Cell Transplantation, Vol. 24, pp. 1981–1997, 2015 0963-6897/15 $90.00 + .

00
Printed in the USA. All rights reserved. DOI: http://dx.doi.org/10.3727/096368914X685302
Copyright Ó 2015 Cognizant Comm. Corp. E-ISSN 1555-3892
www.cognizantcommunication.com

Comparison of Magnetic Intensities for Mesenchymal Stem Cell Targeting

Therapy on Ischemic Myocardial Repair: High Magnetic Intensity Improves
Cell Retention but Has no Additional Functional Benefit
Yunli Shen,*1 Xuebo Liu,*1 Zheyong Huang,† Ning Pei,‡ Jianfeng Xu,† Zheng Li,§ Yunkai Wang,*
Juying Qian,† and Junbo Ge†

*Department of Cardiology, Shanghai East Hospital, Tongji University, Shanghai, China

†Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, Shanghai, China
‡College of Science, Shanghai University, Shanghai, China
§Biomedical Research Center, Zhongshan Hospital, Fudan University, Shanghai, China

Magnetic targeting has the potential to enhance the therapeutic effects of stem cells through increasing reten-
tion of transplanted cells. To investigate the effects of magnetic targeting intensities on cell transplantation, we
performed different magnetic intensities for mesenchymal stem cell (MSC)-targeting therapy in a rat model of
ischemia/reperfusion. Rat MSCs labeled with superparamagnetic oxide nanoparticles (SPIOs) were injected
into the left ventricular (LV) cavity of rats during a brief aorta and pulmonary artery occlusion. The 0.15
Tesla (T), 0.3 T, and 0.6 T magnets were placed 0~1 mm above the injured myocardium during and after
the injection of 1 × 106 MSCs. Fluorescence imaging and quantitative PCR revealed that magnetic targeting
enhanced cell retention in the heart at 24 h in a magnetic field strength-dependent manner. Compared with the
0 T group, three magnetic targeting groups enhanced varying cell engraftment at 3 weeks, at which time LV
remodeling was maximally attenuated, and the therapeutic benefit (LV ejection fraction) was also highest in
the 0.3 T groups. Interestingly, due to the low MSC engraftment resulting from microvascular embolisms, the
0.6 T group failed to translate into additional therapeutic outcomes, though it had the highest cell retention.
Magnetic targeting enhances cell retention in a magnetic field strength-dependent manner. However, too high
of a magnetic intensity may result in microembolization and consequently undermine the functional benefits
of cell transplantation.

Key words: Mesenchymal stem cells; Magnetic targeting intensity; Microembolization; Myocardial
infarction; Rat model

INTRODUCTION delivery of cardiosphere-derived cells (CDCs) (6) or

Cellular therapies are an attractive option as adjunct
therapies for acute or chronic ischemic heart diseases (26).
Intracoronary cell injection is the more clinically relevant
and less invasive setting of an intravascular approach
than intramyocardial injection. However, the therapeutic
effect of intracoronary cell transplantation is extremely
restricted by low cell retention and engraftment in injured
myocardium (10,24).
Magnetic targeting is an emerging approach to augment
cellular therapies by guiding cells to sites of injury using
externally generated magnetic fields and field gradients
(3). Using different magnetic forces, two animal studies
recently tested the magnetically enhanced intracoronary
endothelial progenitor cells (EPCs) (4) in a rat model of
acute ischemia/reperfusion (I/R). In Cheng et al.’s study
(6), the actual working intensity was theoretically esti-
mated to be less than 0.3 Tesla (T) for a 1.3 T magnet
that was placed ~1 cm above the heart, and the magnet
group exhibited greater than a fourfold greater cell reten-
tion. In Chaudeurge et al.’s study (4), a 0.1 T magnet
was implanted into the subcutaneous layer, leading to
an actual working intensity that was far less than 0.1 T,
and magnetic targeting failed to increase cell retention
using RT-PCR quantification. Our previous in vitro study
demonstrated that cell capture efficiency was positively
related to the magnetic flux density between 10 and

Received July 24, 2013; final acceptance September 20, 2014. Online prepub date: November 5, 2014.
1
These authors provided equal contribution to this work.
Address correspondence to Juying Qian, M.D., Shanghai Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, 180 Feng Lin Road,
Shanghai 200032, China. Tel: +86-21-64041990, x10564; Fax: +86-21-64223006; E-mail: juyingqian@126.com or Zheyong Huang, M.D., Shanghai
Institute of Cardiovascular Diseases, Zhongshan Hospital, Fudan University, 180 Feng Lin Road, Shanghai 200032, China. Tel: +86-21-64041990, x12596;
Fax: +86-21-64223006; E-mail: zheyonghuang@126.com
1981
1982
640 mT, and it reached 89.3% with a magnetic flux den-
sity of 640 mT, a magnetic intensity gradient of 38.4 T/m,
and a flow velocity of 0.8 mm/s (14).
Based on the above data, it seems that cell retention
and engraftment can continuously increase with the
enhancement of magnetic force, subsequently translat-
ing into additional cardiac functional and structural ben-
efits. This notion also seems to be evidenced by some
early studies without magnetic targeting, in which the
relationship between injected cell number and func-
tional effect was a positive linear correlation regardless
of intramyocardial injection or intracoronary delivery
(17,22,23,27). However, intracoronary delivery of mes-
enchymal stem cells (MSCs) can induce microvascular
obstruction as has been reported previously (11,29,30),
and little is known about whether microembolisms would
be aggravated with the escalating magnetic intensity and
thereby have an influence on therapeutic outcome of
transplanted cells. Here, we investigated the effects of
magnetic targeting intensities on intraventricular MSC
delivery in a rat I/R model.
MATERIALS AND METHODS
Magnet Cylinder
Three cylindrical NdFeB magnets (Shanghai Yahao
Instrument Equipment Co., Shanghai, China) with 0.15 T,
0.3 T, and 0.6 T (diameter 8 mm) were used in this study.
The magnetic flux density (B) of the magnet surface is
measured using a model 51,662 Leybold Tesla meter
(Beijing Zhuo Sheng Jia Magnetic Technology Co. Ltd.,
Beijing, China). The distribution of the magnetic flux
density was calculated by a finite element analysis.
Rat MSCs, SPIOs, and DiR Labeling
Animal experiments were approved by the Animal Care
and Use Committee of Fudan University and were in com-
pliance with the Guide for the Care and Use of Laboratory
Animals published by the National Academy Press (1).
Femurs of 4-week-old male Sprague–Dawley (SD) rats,
obtained from Shanghai animal administration center,
were excised under sterile conditions. MSCs were cul-
tured from bone marrow plugs explanted from 4-week-old
male SD rats, as previously described (34). All used cells
were harvested by 0.25% trypsin (Gibco, Grand Island,
NY, USA) and 0.038% EDTA (Gibco) when they reached
80–90% confluence at passage 4. MSCs were labeled with
superparamagnetic oxide nanoparticles (SPIOs; 100 mg/
ml; Schering, Berlin, Germany), as previously described
(14). For cell tracking in vivo, cells were labeled with 1,1-
dioctadecyl-3,3,3,3-tetramethyl indotricarbocyanine iodide
(DiR; AAT Bioquest, Inc., Sunnyvale, CA, USA) accord-
ing to the manufacturer’s protocol. Labeled cells are here-
after called SPIOs-DiR-MSCs or DiR-MSCs for brevity.
To evaluate the labeling efficacy, Prussian blue staining
SHEN ET AL.
(potassium ferrocyanide and HCl solution from Jueshen
Bio-Technology Co. Ltd., Shanghai, China) and electron
microscopy (Phillips CM120, Philips FEI, Eindhoven,
Netherlands) were undertaken to indicate the presence and
localization of intracellular iron particles, and DiR stain-
ing was observed by fluorescence microscopy using the
XF112-Omega optical filter after cell nuclei were coun-
terstained with 4¢,6-diamidino-2-phenylindole (DAPI;
Sigma-Aldrich, St. Louis, MO, USA) (16).
Effects of SPIO Labeling on MSC
Differentiation Capacity
Our previous study demonstrated that SPIO labeling
had no significant effect on the proliferation and viabil-
ity of MSCs (16). Here we further determined the dif-
ferentiation capacity of SPIOs-MSCs. The MSCs and
SPIOs-MSCs were, respectively, cultured in adipogenic
(SR811D250, Amsbio, Abingdon, UK) or osteogenic
(SR417D250, Amsbio) media for 21 days with media
replenishment every 3 days. To demonstrate adipogenic
differentiation, the cells were stained with 0.3% Oil red O
(O0625; Sigma-Aldrich) in isopropanol for 30 min and
rinsed with PBS. For osteogenic differentiation, the cells
were stained with 1% Alizarin red (500-4; RiccaChem,
Arlington, TX, USA) for 15 min (2).
To determine the myocardium-like cell differentiation,
the MSCs and SPIOs-MSCs were cultured in culture
medium-added 5-aza-2¢-deoxycytidine (5-azaC; Sigma-
Aldrich) for 24 h. Incubation then continued in complete
medium lacking 5-azaC, with medium changes every 3 days.
Undifferentiated cells and normal mature rat cardiomyo-
cytes were, respectively, used as negative and positive
controls. Total RNA was extracted from cultured cells
using TRIzol reagent (Invitrogen, Carlsbad, CA, USA)
according to the manufacturer’s protocol. RT-PCR prim-
ers (synthesized by Sangon, Shanghai, China) were as fol-
lows: GATA-4, 5¢-CTG TCA TCT CAC TAT GGG CA-3¢
and 5¢-CCA AGT CCG AGC AGG AAT T T-3¢; a-actin,
(F) 5¢-TCC TTT ATC GGT ATG GAG TCT G-3¢ and
(R) 5¢-TGA TCT TGA TCT TCA TGG TGC T-3¢; cTnT,
5¢-AGA GGA CTC CAA ACC CAA GC-3¢ and 5¢-ATT
GCG AAT ACG CTG CTG TT-3¢; Cx43, 5¢-TCC TTG
GTG TCT CTC GCT TT-3¢ and 5¢-GAG CAG CCA TTG
AAG TAG G C-3¢; GAPDH, 5¢-GAC ATG CCG CCT
GGA GAA AC-3¢ and 5¢-AGC CCA GGA TGC CCT
TTA GT-3¢. Transcriptional expressions of GATA-4, cTnT,
a-actin, and Cx43 genes from the MSCs and SPIOs-MSCs
were determined by real-time PCR according to the manu-
facturer’s instructions. Transcript levels were standardized
to the corresponding rat GAPDH level.
In Vitro Magnetic Capture of Flowing SPIOs-MSCs
To test the magnetic responsiveness of MSCs labeled
with SPIOs (SPIOs-MSCs), the accumulation of SPIOs-
MAGNETIC INTENSITY FOR STEM CELL TARGETING
MSCs was observed, while cells flowed through a tube
served as a model of blood vessels in a magnetic field. A
total of 20 ml SPIOs-MSCs suspension, at a concentra-
tion of 5 × 104 cells/ml, was placed in a 50-ml syringe
(Shanghai Boguang Biotechnology Co. Ltd., Shanghai,
China) and flowed through a quartz tube (ID 2.3 mm,
OD 4.3 mm, length 20 cm) (Shanghai Puya Quartz Glass
Factory, Shanghai, China). Cell suspension circulated
at flows of 5 mm/s to mimic circulatory conditions in
animals and human beings. The aforementioned 0.15-T
magnet cylinder was placed tightly at the midsegment of
the tube. The cell suspension flowed through the mag-
net field was collected in a centrifuge tube (Shanghai
Boguang Biotechnology Co. Ltd.) 1#. The remaining
cells within the quartz tube were washed using PBS and
collected in centrifuge tube 2#. The numbers of SPIOs-
MSCs in centrifuge tube 1# and 2# were counted using a
counting plate and referred to quantity of captured cells
(Q1) and quantity of uncaptured cells (Q2). The capture
Q1
efficiency (CE) was calculated by CE = × 100%.
Q1 + Q2
Then, the effects of the 0.3-T and 0.6-T magnets on the
accumulation of SPIOs-MSCs in turn were observed.
The magnetic responsiveness of MSCs unlabeled with
SPIOs was tested according to the method mentioned
above. All experiments were performed in six copies for
each condition.
Animal Model and Magnetic Targeting
To make our experiment more relevant to reperfused
acute myocardial infarction (AMI), a rat I/R model was
developed in female SD rats (150–200 g). Rats were
anesthetized with ketamine (100 mg/kg, IP) (Beijing
Zizhu Pharmaceutical Co. Ltd., Beijing, China) and
xylazine (10 mg/kg, IP) (Shanghai Ruicong Laboratory
Equipment Co. Ltd., Shanghai, China) and were venti-
lated with a rodent ventilator using room air at 80 breaths
per minute. Female SD rats (n = 236 total), obtained from
Shanghai animal administration center, underwent a left
thoracotomy in the fourth intercostal space under gen-
eral anesthesia. The heart was exposed, and myocardial
infarction was produced by a 90-min ligation of the left
anterior descending coronary artery (LAD) using a 7-0
silk suture (Unik Surgical Sutures Mfg. Co. Ltd., Suzhou,
China). The suture was then released to allow coronary
reperfusion. Twenty minutes later, cells were injected into
the left ventricle cavity during a 5-s temporary aorta and
main pulmonary artery occlusion, as previously described
(18,19). Cell delivery was confirmed by the temporary
observation of bradycardia and epicardial blanching.
For magnetic targeting, three cylindrical NdFeB mag-
nets with 0.15 T, 0.3 T, and 0.6 T were separately placed
above the injured myocardium during and after cell injec-
tion for 10 min, which was determined to be the optimal
1983
time (5,6). The 217 survivors were randomized into six
groups according to the strength of the magnetic field
(n = 37 for 0.3 T group, n = 36 for other groups): 0 T group,
0.15 T group, 0.3 T group, 0.6 T group, MSC group, and
PBS group. All magnets were close to the target site, that
is, within 0–1 mm, during and after infusion in the three
magnetic targeting groups. Except for the PBS group,
which was intraventricularly injected with vehicle (PBS)
only, the other treatment groups were intraventricularly
injected with 1 × 106 SPIOs-DiR-MSCs (0 T, 0.15 T, 0.3 T,
and 0.6 T group) or DiR-MSCs (MSC group). All animals
received humane care in compliance with the Guide for
the Care and Use of Laboratory Animals.
Fluorescence Imaging
After assessment of cardiac function using trans-
thoracic echocardiography, six animals were randomly
selected from each treatment group and were eutha-
nized 24 h after cell injection for fluorescence imaging.
Extensive PBS washing was performed to remove any
cells adherent to the surface of organs. Hearts, lungs, livers,
spleens, and kidneys were placed in a Carestream In-Vivo
Multispectral Imaging System FX PRO (Carestream
Health, Toronto, Canada) to detect DiR fluorescence under
748 nm of excitation and 780 nm of emission. Exposure
time was set at 3 s and kept the same during the entire
imaging session. Organs from the PBS group (i.e., the
animals receiving normal saline) were also imaged as
controls for background noise. Fluorescence signals
(photon/s) from a fixed region of interest (ROI) were
measured as described (36).
Quantification of Engraftment by Real-Time PCR
Quantitative PCR was performed 24 h (after fluo-
rescence imaging and assessment of cardiac function),
10 days, and 3 weeks (after fluorescence imaging and
assessment of cardiac function) after cell injection in six
animals from each cell-injected group to quantify cell
retention/engraftment. We injected MSCs from male donor
SD rats into the left ventricle of female recipients, uti-
lizing the sex-determining region Y (SRY) gene located
on the Y chromosome as a detection target (28). The
whole heart was harvested, weighed, and homogenized.
Genomic DNA was isolated from aliquots of the homoge-
nate corresponding to 12.5 mg of myocardial tissue using
commercial kits (DNA Easy minikit, Qiagen, Hilden,
Germany). The TaqMan® assay (Applied Biosystems,
Carlsbad, CA, USA) was used to quantify the number of
transplanted cells with the rat SRY gene as a template
[forward primer: 5¢-GAG CTT TGG GAG CAG TGA
C-3¢ (TM 55.1), reverse primer 5¢- ATG AGG CTG ATA
TTT ATA GTT TGG-3¢ (TM 51.5), Taq Man probe: 6
FAM CAA CAG AAT CCC AGC ATG CAG AAT TCA
G TAMRA; Applied Biosystems]. A standard curve was
1984
generated with multiple dilutions of genomic DNA iso-
lated from male hearts to quantify the absolute gene copy
numbers. All samples were spiked with equal amounts
of female genomic DNA as a control. The copy number
of the SRY gene at each point of the standard curve was
calculated with the amount of DNA in each sample and
the mass of the rat genome per cell. For each reaction,
50 ng of template DNA was used. Real-time PCR was
performed using an Applied Biosystems 7900 HT Fast
real-time PCR System. All experiments were performed
in triplicate. Cell numbers per milligram of heart tissue
and percentages of retained cells of the total injected cells
(1 × 106) were calculated.
Assessment of Cardiac Function
Postinfarct cardiac function was evaluated by transtho-
racic echocardiography using a Vevo 770 high-resolution
imaging system (Visual Sonics, Toronto, Canada) with a
17.5-MHz probe 24 h and 3 weeks after I/R and treat-
ment (n = 13 for the 0.3 T group, n = 12 for other groups).
After the induction of light general anesthesia, hearts
were imaged in 2D and M-mode. The recordings were
obtained from the parasternal long axis view at the level
of the greatest LV diameter. The LV internal end-diastolic
diameter (LVIDd) and LV internal end-systolic diameter
(LVIDs) were measured from M-mode recordings. The
LV ejection fraction (LVEF) was calculated as follows:
LVEF (%) = [(LVIDd)3 − (LVIDs)3]/(LVIDd)3 × 100. All
echocardiographic measurements were averaged from at
least five separate cardiac cycles.
Immunohistochemistry and Morphometric
Heart Analysis
At 72 h after cell injection, six representative recipient
hearts from each group embedded in paraffin were cut
into 5-µm slices, which were stained with Prussian blue
(Shanghai HuaYi Bio Technology Co. Ltd., Shanghai,
China). The representative hearts were harvested and fro-
zen in OCT compound (Leica JUNG, Wetzlar, Germany)
72 h after transplantation and 3 weeks subsequent to
Table 1. Study Protocol and Rat Numbers Used in Each Experiment
24 Hours 72 Hours
Heart
Fluorescence/ Prussian
PCR for SRY Blue Staining
PBS group 6 6
MSC group 6 6
0 T group 6 6
0.15 T group 6 6
0.3 T group 6 6
0.6 T group 6 6
Twelve animals from each group (except for 13 animals in 0.3 T group) underwent echocardiography at 24 h and 21 days after injection, respectively.
SRY, sex-determining region Y gene; PBS, phosphate-buffered saline (control); MSC, mesenchymal stem cell; T, Tesla (magnetic field strength).
SHEN ET AL.
echocardiography of the rat. Cryostat sections (8 mm
thickness) were obtained for immunofluorescence stain-
ing to evaluate cardiomyocyte or endothelial differentia-
tion of injected MSCs. Potential transdifferentiation of
myocardium-like cells from implanted MSCs was veri-
fied by antibody immunostaining for troponin T (cTnT).
Briefly, frozen tissue sections were fixed in acetone at
4°C for 10 min and incubated separately with a mouse
anti-rat cTnT (TnT, cardiac isoform Ab-1; Labora-
tory Vision Corp, Chicago, IL, USA) diluted 1:100 for
60 min at room temperature. After washing with PBS
solution, sections were incubated with a goat anti-mouse-
conjugated FITC IgG (Molecular Probes, Inc., Eugene,
OR, USA) diluted 1:200 for cTnT. Goat anti-rat CD31
(BD Biosciences, San Diego, CA, USA) was diluted
1:100 to check for evidence of angiogenesis. Secondary
antibody of donkey anti-goat IgG conjugated with FITC
(Molecular Probes) diluted 1:200 was used. Cell nuclei
were counterstained with DAPI. Capillary density was
also assessed by staining with anti-CD31 antibody as pre-
viously described (31). A total of 10 visual fields where a
cross-section of capillaries was clearly visible were ran-
domly selected in the peri-infarct zone, and the number
of capillaries was counted under 200× magnification by a
Leica Fluorescence microscope (Leica DM LB2).
For morphometric analysis, representative recipient
hearts in each group were harvested at 3 weeks (after
cardiac function assessment) and embedded in paraffin.
Masson’s trichrome staining was performed as described
by the manufacturer’s instructions [HT15 Trichrome
Staining (Masson) Kit; Sigma-Aldrich]. To determine
the infarct size, the average ratio of infarct size per LV
was measured for all paraffin slides after staining with
Masson’s trichrome as previously described (21). All
analyses were performed in a blinded manner.
Statistical Analysis
Animal numbers used in each experiment are shown
in Table 1. All data are shown as mean ± standard devia-
tion (SD). The difference between mean values was
10 Days 21 Days
Cell PCR Masson Staining/
Fluorescence for SRY Immunofluorescence PCR for SRY
6 6 6 6
6 6 6 6
6 6 6 6
6 6 6 6
6 6 7 6
6 6 6 6
MAGNETIC INTENSITY FOR STEM CELL TARGETING 1985

determined with one-way analysis of variance (ANOVA)
and Bonferroni post hoc test. The correlation analysis was
based on the Pearson correlation coefficient. Statistical
significance was set at p < 0.05.
RESULTS
Labeling of Rat MSCs With SPIOs and DiR
The fourth passage of MSCs was characterized by
fusiform or spindle-shaped morphology (Fig. 1A). The
SPIOs-DiR-MSCs had a strong red fluorescence sig-
nal (Fig. 1B). The Prussian blue staining of the MSCs
showed intracytoplasmic iron inclusions as dense blue-
stained vesicles, and the magnetic nanoparticles were
distributed evenly around the nucleus of the MSCs as
a spherical shell (Fig. 1C). The labeling efficiency was
approximately 100% reproducible using a very low con-
centration of iron oxide (50 µg/ml). Transmission elec-
tron microscopy of labeled cells indicated the presence
of anionic magnetic nanoparticles exclusively in polydis-
perse vesicles in the cytoplasm, and no obvious change in
the ultramicrostructure was observed (Fig. 1D).
Characterization of SPIOs-MSCs
To determine the adipogenic and osteogenic differ-
entiation, the MSCs and SPIOs-MSCs were cultured for
21 days in media that promote differentiation into adi-
pogenic and osteogenic lineages. As shown in Figure 2A
and B, both MSCs and SPIOs-MSCs differentiated into
the two lineages, as demonstrated by staining for Oil
red O and Alizarin red, respectively. Following myo-
cardial differentiation, real-time PCR showed that myo-
cardial genes GATA-4, cTnT, a-actin, and Cx43 were
both detected in differentiated MSCs and SPIOs-MSCs
(all p > 0.05) (Fig. 2C). The expression of these myocar-
dial genes was comparable in differentiated MSCs and
SPIOs-MSCs (Fig. 2D). The results from RT-PCR sug-
gested that low concentration of SPIO labeling did not
affect MSC differentiation.
In Vitro Theoretical Analysis of the Magnetic Targeting
The magnetic field lines around the magnet were
mapped by point-by-point magnetic field measurements
using a model 51662 Leybold Tesla meter. The center
magnetic flux densities of three cylindrical magnet poles
were 0.15 T, 0.3 T, and 0.6 T, and the peripheral strengths
of the corresponding magnetic poles were 0.40 T, 0.56 T,
and 0.93 T, respectively. As shown in Figure 3A–C, the
magnetic field strength gradually declines with the short-
ened distance from periphery to center of the magnetic
pole in the horizontal direction. Line 1 and line 2,

Figure 1. SPIOs and DiR labeling of rat MSCs. (A) The fourth passage of MSCs used in the present study. (B) MSCs were labeled
with DiR, and cell nuclei were counterstained with DAPI. (C) Prussian blue staining of MSCs labeled with SPIOs on cover glass.
(D) Transmission electron microscopy demonstrated the presence of numerous iron-containing vesicles in the cytoplasm of MSCs.
SPIOs, superparamagnetic oxide nanoparticles; DiR, 1,1-dioctadecyl-3,3,3,3-tetramethyl indotricarbocyanine iodide; MSCs, mesen-
chymal stem cells; DAPI, 4¢,6-diamidino-2-phenylindole.
1986 SHEN ET AL.

Figure 2. Differentiation capacity of SPIOs-MSCs. (A) Oil red O staining for adipogenic differentiation. (B) Alizarin red staining for
osteogenic differentiation. (C) Real-time PCR for myocardial gene GATA-4, cTnT, a-actin, and Cx43. (D) Quantitative gene expres-
sion analysis showed no difference between MSCs and SPIOs-MSCs, suggesting low concentration of SPIO labeling did not affect
MSCs differentiation. NC, negative control; PC, positive control.

respectively, represent magnetic field lines from the
center and periphery of the magnetic pole in the vertical
direction (Fig. 3D). According to the principle of mag-
netic attenuation, when the magnetic pole radius is 4 mm,
the density of magnetic flux is attenuated by 47%, 43%,
and 40% at 2 mm from the magnetic poles with 0.15 T,
0.3 T, and 0.6 T and is further decreased by 83%, 80%,
and 76% at 6 mm from the three magnets, respectively
(Fig. 3E–G). Thus, it is critical not only to overcome the
magnetic attenuation but also to make magnetic field
lines cover the target site for administering a specific
strength of magnetic targeting. We controlled the move-
ment ranges of magnets with the heartbeat to within
1 mm to maximally overcome magnetic attenuation. The
actual measured working strengths of the magnets with
0.15 T, 0.3 T, and 0.6 T were at least 0.12 T, 0.23 T, and
0.49 T, respectively. All magnets used here were 8 mm in
diameter and approximately the size of a MI in rats. We
 MAGNETIC INTENSITY FOR STEM CELL TARGETING

Figure 3. Theoretical analysis of magnetic fields. (A–C) Distribution of magnetic field lines from three magnets. In the horizontal direction, the magnetic field strength gradually
declines along the radius from the periphery to the center. (D–G) Schematic diagram of magnetic attenuation in the vertical direction. Lines 1 and 2 represent magnetic decay curves
of central and peripheral of magnetisms, respectively, in the vertical direction.
1987
1988 SHEN ET AL.

managed to cover the infarct area and infarct border zone
with the strongest area of the magnetic field, thus ensur-
ing target homing of magnetic-responsive cells.
Magnetic Capture of Flowing SPIOs-MSCs
The flowing SPIOs-MSCs were substantially attracted
to the site where the magnetic pole was positioned.
The flowing SPIOs-MSCs in the absence of an applied
magnetic field demonstrated minimal accumulation
(0.26% ± 0.04%), while the CEs reached 42.92% ± 4.76%,
70.53% ± 6.53%, and 80.2% ± 4.79% when the magnetic
intensities were 0.15 T, 0.3 T, and 0.6 T, respectively
(all p < 0.001 among groups) (Fig. 4A–C). The capture
efficiency of cells increased as the magnetic intensity
enhanced, indicating that the flowing SPIOs-MSCs
can be significantly magnetically attracted in a magnetic
flux density-dependent manner. In contrast, the flow-
ing unlabeled MSCs exposed to the varying intensities
of magnetic fields only showed minimal accumulation
(0.22% ± 0.04%, 0.21% ± 0.02%, 0.21% ± 0.02% and
0.22% ± 0.03% for 0 T, 0.15 T, 0.3 T, and 0.6 T, respec-
tively; all p > 0.05 among groups), indicating that MSCs
without SPIOs were not magnetically responsive.
Magnetic Strength Influences Cell Distribution
After Retention
Because vascular-delivered cells reportedly translocate
into the parenchyma after 48–72 h (29), a subpopula-
tion of animals was histologically examined at 72 h. Rep-
resentative hearts were cryosectioned and analyzed for
covisualization of CD31+ blood vessels (green) and DiR-
labeled MSCs (red). Cells were found residing in the
infarct area and peri-infarct border zone in the groups
treated with SPIOs-DiR-MSCs or DiR-MSCs. Not sur-
prisingly, the numbers of DiR-MSCs were indistinguish-
able in the 0 T and MSC groups (p > 0.05) (Fig. 5A, B).

Figure 4. Magnetic concentration of the flowing SPIOs-MSCs. (A) A schematic illustration of the microfluidic system. The SPIOs-
MSCs were injected into the tube, and the flow rate was controlled using a syringe pump. Cells were captured by use of three NdFeB
magnets located beside the tube. (B) Representative images of cell accumulation in the segment close to the magnet at velocities of
5 mm/s. (C) The efficiency of magnetically capturing flowing SPIOs-MSCs with different magnetic intensities. (#p < 0.001 vs. the 0 T
group; Dp < 0.001 vs. the 0.15 T group; ☆p < 0.01 vs. the 0.3 T group).
MAGNETIC INTENSITY FOR STEM CELL TARGETING 1989

Three magnetic targeting groups exhibited more SPIOs-
DiR-MSCs compared to either the 0 T group or the MSC
group (all p < 0.001) (Fig. 5A–E). DiR fluorescence from
MSCs suggested that the 0.15 T, 0.3 T, and 0.6 T groups
demonstrated 2.18-, 3.95-, and 2.31-fold cell distribution,
respectively, outside vascular structures relative to the 0 T
group (all p < 0.001, respectively) (Fig. 5F). Interestingly,
comparable results were found in the 0.6 T and 0.15 T
groups (p > 0.05).
Meanwhile, microemboli were detected in the groups
treated with MSCs (Fig. 6A, B). The percentage of
blocked vessels was comparable in the MSC and 0 T
groups (2.93 ± 1.12% vs. 3.16 ± 1.36%, p > 0.05), indicat-
ing that MSCs can induce microvascular obstruction as
has been reported previously (13,30). More importantly,
the percentage of blocked vessels was 4.3% ± 0.98%,
8.24% ± 1.12%, and 15.12% ± 2.09% when the magnetic
intensities were 0.15 T, 0.3 T, and 0.6 T, respectively,
indicating that the external magnetic field may exacer-
bate magnetically responsive MSCs obstructed in micro-
vascular, and the microembolisms further worsened with
the escalating magnetic intensity (Fig. 6C).
The Relationship Between Magnetic Strength and
Short-Term Cell Retention
Fluorescence imaging of hearts excised 24 h after cell
infusion revealed more fluorescence signals in hearts
from the three magnetic targeting groups than the 0 T

Figure 5. Microembolization at 72 h after intraventricular infusion (n = 6 for each group). (A–E) Representative fluorescence image
of heart sections from MSC group (A), 0 T group (B), 0.15 T group (C), 0.3 T group (D), and 0.6 T group (E). CD31 antibody (green)
stained for detecting blood vessels. MSCs were visualized by red fluorescence, and cell nuclei were counterstained with DAPI.
(F) MSC numbers from 10 randomly selected high-power fields (five from the infarct area and five from the border zone) in each group
were quantified by fluorescence microscopy. Data are mean ± SD. (#p < 0.001 vs. the MSC group or the 0 T group; Dp < 0.001 vs. the
0.15 T group; ☆p < 0.001 vs. the 0.3 T group). HPF, high power field.
1990 SHEN ET AL.

Figure 6. Microembolization at 72 h after intraventricular infusion (n = 6 for each group). (A) Representative heart frozen sections
were stained for CD31 antibody (green) to detect blood vessels. SPIOs-DiR-MSCs were visualized by red fluorescence, and cell nuclei
were counterstained with DAPI. The 0.6 T groups showed blood vessel completely blocked by cell clumps, whereas the 0 T, 0.15 T,
and 0.3 T groups exhibited that blood vessels containing cells were still patent. (B) Prussian blue staining confirmed that blood vessels
were blocked by cell clumps in the 0.3 T and 0.6 T groups (arrows), but most blood vessels containing cells were still patent in the 0 T
and 0.15 T groups (arrows). (C) Quantitative analysis of blocked vessels by frozen sections stained with CD31 antibody. (#p < 0.001
vs. the MSC group or the 0 T group; Dp < 0.001 vs. the 0.15 T group; ☆p < 0.001 vs. the 0.3 T group).

and MSC groups. The enhanced fluorescence from three
magnetic targeting groups was dependent on magnetic
strength. As a negative control, excised hearts from the
PBS group were also imaged. No signals were detectable
(Fig. 7A, B).
To compare off-target migration, lungs, livers, spleens,
and kidneys from the same animals were also harvested
to detect fluorescence signals of transplanted MSCs.
Not surprisingly, fluorescence signals were detectable
in these organs, but less so in the organs from three mag-
netically targeted groups than in those from the MSC
and 0 T groups (Fig. 7C). Moreover, there is a trend
that fluorescence signals in the off-target organs from
three magnetically targeted groups decreased with the
enhancement of strength of heart magnetic targeting
(Fig. 7C).
Quantitative PCR for the male-specific SRY gene
was performed 24 h after cell infusion. Consistent with
the fluorescence imaging results, magnetic targeting
enhanced short-term cell retention in a magnetic strength-
dependent manner: the 0.15 T, 0.3 T, and 0.6 T groups
exhibited 1.45-, 2.76-, and 3.86-fold greater cell reten-
tion rates, respectively, than the 0 T group (all p < 0.001)
(Fig. 7D). The relationship between magnetic intensity
and retained cell numbers is a positive linear correlation
(r = 0.983, p = 0.017).
MAGNETIC INTENSITY FOR STEM CELL TARGETING 1991

Figure 7. Short-term cell retention and long-term cell engraftment in hearts and other organs. (A) Representative fluorescence imag-
ing of hearts harvested 24 h after cell injection. (B) Quantificative analysis of fluorescence intensities. (*p < 0.001 vs. the PBS group;
#p < 0.001 vs. the MSC group or the 0 T group; Dp < 0.001 vs. the 0.15 T group; ☆p < 0.001 vs. the 0.3 T group). (C) Cell retention in
other organs by fluorescence imaging 24 h after cell injection. (#p < 0.001 vs. the MSC group or the 0 T group; Dp < 0.001 vs. the 0.15 T
group; ☆p < 0.001 vs. the 0.3 T group). (D) Cell detection 24 h, 10 days, and 3 weeks after injection in the heart by quantitative PCR
of the male-specific SRY gene (n = 6 for each group).

Different Magnetic Strengths Induce Varying Long-Term
Cell Engraftment
To examine the effects of magnetic targeting on long-
term engraftment, quantitative PCR for the male-specific
SRY gene was performed 10 days and 3 weeks after cell
infusion. Consistent with previous findings (4,6,19,20),
quantitative PCR revealed that the five groups treated
with MSCs experienced a substantial drop in surviving
cells 10 days after cell infusion. The surviving cells fur-
thered reduced 3 weeks after cell infusion; however, the
three magnetic targeting groups still exhibited enhanced
cell engraftment relative to the 0 T group (all p < 0.001)
(Fig. 7D), which is consistent with previous findings
(5,6), indicating that magnetic targeting can enhance
MSC homing and engraftment. At 3 weeks after cell
infusion, the 0.15 T, 0.3 T, and 0.6 T groups exhibited
1.88-, 3.06-, and 1.90-fold greater cell engraftment rates,
respectively, than the 0 T group (Fig. 7D). Interestingly,
cell engraftment rates in the 0.6 T group were lower than
the 0.3 T group (p < 0.001) (Fig. 7D). Compared with the
0.15 T group, the 0.6 T group exhibited comparable cell
engraftment rates (p > 0.05) (Fig. 7D).
Magnetic Strength Affects Therapeutic Benefits of Cell
Transplantation and Left Ventricular Remodeling
To investigate functional outcomes mediated by mag-
netic targeting, LVEF was assessed by echocardiography
at baseline (24 h after I/R and treatment) and for a sub-
sequent 3 weeks. The 0.6 T group had the lowest LVEF
at baseline, but LVEF did not differ between treatment
groups (Fig. 8A), indicating a comparable degree of ini-
tial injury. Over the next 3 weeks, LVEF declined progres-
sively in the PBS group, but it did not decline in the four
MSC-treated groups. Both the 0.15 T and 0.3 T groups
exhibited better therapeutic outcome, with LVEF superior
to the 0 T group (all p < 0.001) (Fig. 8A) at 3 weeks. Not
1992 SHEN ET AL.

Figure 8. Functional benefits of cell transplantation. (A) LVEF measured by echocardiography 24 h and 3 weeks after cell injection
(n = 13 for 0.3 T and n = 12 for other groups). (B) Changes in LVEF from 24 h to 3 weeks in each group. (*p < 0.001 vs. the PBS group;
#p < 0.001 vs. the MSC group or the 0 T group; Dp < 0.001 vs. the 0.15 T group; ☆p < 0.001 vs. the 0.3 T group). (C–D) Changes in
LVEF are plotted against changes in LVIDs (C) and LVIDd (D). Change reflects the difference between parameters measured 24 h
and 3 weeks after therapy. LVEF, left ventricular ejection fraction; LVIDs, left ventricular internal end-systolic diameter; LVIDd, left
ventricular internal end-diastolic diameter; DLVEF, Changes in LVEF from 24 h to 3 weeks.

surprisingly, the 0.3 T group had a higher LVEF than the
0.15 T group (p < 0.001) (Fig. 8A). Notably, LVEF was
comparable among the 0.6 T, 0 T, and MSC groups (all
p > 0.05) (Fig. 8A). To facilitate comparisons, we calcu-
lated the treatment effect, that is, the change in LVEF at
3 weeks relative to baseline, in each group.
The PBS group had a negative treatment effect, with
LVEF decreasing over time. In contrast, the 0.15 T and
0.3 T groups exhibited a sizable positive treatment effect
that was greater than the 0 T and 0.6 T groups (Fig. 8B).
The similar changes were also observed in LVIDd and
LVIDs. Compared with the MSC, 0 T, and 0.6 T groups,
the 0.15 T group exhibited the greater reduction in LVIDd
and LVIDs (0.21 mm and 0.15 mm, 0.19 mm and 0.13 mm,
0.21 mm and 0.14 mm vs. 0.59 mm and 0.40 mm, respec-
tively, all p < 0.001). More importantly, the 0.3 T group
exhibited the greatest reduction in LVIDs (1.4 mm) and
LVIDd (0.90 mm) among three magnetic targeting groups
(all p < 0.001). The percentage of change in the LVEF at
3 weeks, as a function of the LVIDs (Fig. 8C) or LVIDd
(Fig. 8D) percentage of change indicated that appropri-
ate intensity of magnetic targeting mediating cell therapy
extended the therapeutic benefit of cell therapy for car-
diac function and structure.
Morphometry at 3 weeks showed more infarct size in
the PBS group than the five MSC-treated groups. The
0.15 T and 0.3 T groups showed significantly smaller
infarct size than the 0 T group and the MSC group. Fur-
thermore, the infarct size was 4.7% lower in the 0.3 T
group than the 0.15 T group (p < 0.001) (Fig. 9A, B).
Again, LV infarct size was indistinguishable among the
0.6 T, 0 T, and MSC groups (p > 0.05) (Fig. 9A, B).
Different Intensities of Magnetically Targeted Cell
Delivery Induce Varying Angiogenesis
To determine the mechanisms underlying the ben-
eficial effects of cell transplantation, we investigated the
effects of cell transplantation on angiogenesis in the post-
I/R hearts. At 3 weeks after cell transplantation, immu-
nofluorescence staining for CD31 antibody indicated
MAGNETIC INTENSITY FOR STEM CELL TARGETING 1993

Figure 9. Left ventricular remodeling. (A) Representative Masson’s trichrome stained myocardial sections from a subgroup of ani-
mals 3 weeks after treatment (n = 7 for 0.3 T and n = 6 for other group). (B) Quantitative analysis of fibrosis size. (*p < 0.001 vs. the PBS
group; #p < 0.001 vs. the MSC group or the 0 T group; Dp < 0.01 vs. the 0.15 T group; ☆p < 0.001 vs. the 0.3 T group).

significant angiogenesis in MSC-treated hearts, with
more CD31-expressing capillaries being present in the
peri-infarct region of MSC-treated groups compared with
the PBS group (all p < 0.05) (Fig. 10A, B). Quantitative
evaluation of CD31-positive capillary numbers indicated
that the PBS group exhibited the lowest capillary den-
sity. In contrast, both the 0.15 T and 0.3 T groups had
more capillary density than the 0 T and 0.6 T groups (all
p < 0.01) (Fig. 10A, B). Not surprisingly, the 0.3 T group
had the most capillary density. Also, no significant differ-
ence was found among the 0.6 T, 0 T, and MSC groups
(p > 0.05) (Fig. 10B). However, few cells stained positive
for both the endothelial cell marker CD31 or the cardio-
myocyte marker TnT and DiR in the five MSC-treated
groups. These data suggest that transplanted MSCs
may promote angiogenesis through a paracrine effect,
contributing to the observed cellular therapy-associated
beneficial effects.
DISCUSSION
To our knowledge, this is the first study that compares
the impact of different strengths of magnetic targeting
on the therapeutic outcomes of intraventricular deliv-
ery of MSCs in a rat model of ischemia/reperfusion. We
found that magnetic targeting enhanced cell retention in
a magnetic intensity-dependent manner, but “too high”
of a strength can worsen embolic injury, thus undermin-
ing the therapeutic effects of cell transplantation. A num-
ber of animal studies and clinical trials have confirmed
that transplantation effect was positively related to cell
dose. Pouzet et al. reported that a fivefold difference
in the number of transplanted skeletal myoblasts (SM,
1994 SHEN ET AL.

Figure 10. Angiogenesis 3 weeks after cell infusion. (A) Frozen sections of representative hearts were stained for CD31 antibody
(green) to detect capillary densities in the peri-infarct region 3 weeks after MSC transplantation. (B) Quantitative analysis of CD31-
containing capillaries in the peri-infarct region (n = 7 for 0.3 T and n = 6 for other group). (*p < 0.05 vs. the PBS group; #p < 0.05 vs. the
MSC group or the 0-T group; Dp < 0.01 vs. the 0.15-T group; ☆p < 0.001 vs. the 0.3-T group).

1 ´ 106 vs. 5 ´ 106) caused a twofold difference in LVEF
(22). Tambara et al. found that a large number of trans-
planted neonatal SM can replace the infarcted myocar-
dium with reverse LV remodeling in infracted rat hearts
(27). Clinical trials (17,23,32) and meta analysis (7) also
demonstrated that heart function in patients with AMI
was related to intracoronary transplanted cell dose. Our
previous study found that repeated bone marrow mono-
nuclear cell administration can significantly improve LV
function, compared with a single infusion in patients
with large AMI (35). Despite different cell types and
delivery routes and different heterogeneity evaluations,
these results demonstrated that, in patients with AMI, the
effects of cell transplantation are dependent on the num-
ber of cells. This finding is the theoretical basis of cell
transplantation guided by magnetic targeting. Previous
studies have found that magnetic targeting can enhance
cell homing and improve therapeutic effects irrespective
of delivery route (5,6).
We found that the 0.15 T, 0.3 T, and 0.6 T groups
exhibited 1.45-, 2.76-, and 3.86-fold greater cell retention
rates, respectively, than the 0 T group. Cheng et al. (6)
MAGNETIC INTENSITY FOR STEM CELL TARGETING
reported the magnet group exhibited more than fourfold
greater cell retention rates than the cardiosphere-derived
cell (CDC) group. Chaudeurge et al. (4) implanted a 0.1 T
magnet (7 mm diameter, 5 mm thickness) subcutane-
ously, leading to cell retention between groups failing to
be statistically significant using RT-PCR quantification.
We speculate that the different results may be attributed
to the difference in magnetic intensities and cell types.
To our knowledge, there are several factors affecting the
magnetic attraction of transplanted cells. First, the mag-
netic strength and the magnetic exposure time must be
taken into consideration. Notably, the magnet intensity is
not equal to the actual working strength, which is limited
by magnetic attenuation. To overcome magnetic attenua-
tion and obtain maximal working magnetic strength, the
distance from the magnetic pole to the target site should
be as short as possible. Second, it is critical that the dis-
tribution of magnetic field lines must cover the target
site. Third, the iron content and the magnetic responsive-
ness of the transplanted cells may also be an important
factor. In the present study, the distance from magnet to
target lesion was 0~1 mm to maximally overcome mag-
netic attenuation; moreover, although the three magnets
we used had different strengths, their surface areas were
close to the size of a rat MI, ensuring the magnetic target-
ing effect.
Surprisingly, we found the 0.6 T group had the highest
cell retention, but at 3 weeks cell engraftment was indis-
tinguishable relative to the 0.15 T group; moreover, thera-
peutic benefits had no significant difference compared to
the 0 T group. What led to the paradoxical phenomenon
between cell retention and therapeutic effects? We found
more microembolization in the 0.6 T group at 72 h than
the 0.15 T and 0.3 T groups. We speculate that the para-
doxical result may be associated with severer microem-
bolization (15). Owing to more serious microembolisms,
although the 0.6 T group had the highest cell retention, the
retained cells did not represent effectively homing cells.
Consequently, the result did not translate into enhanced
cell engraftment. Moreover, these microembolisms may
cause new myocardial injury or deteriorate original injury,
partly offsetting the therapeutic benefit of homing cells
and eventually resulting in comparable functional effects
in the 0.6 T and 0 T groups. Basically, the cell transplanta-
tion effect is dependent on the effective homing cell num-
ber rather than retained cell number.
The intracoronary injection of MSCs has been proposed
as a potential therapeutic option to repair an ischemic- or
infarct-damaged heart. However, because MSCs are large
(rounded up cells are ~22 to 25 µm in diameter), an important
consideration of this approach is the potential of these cells
to induce myocardial damage via microvascular obstruc-
tion, as previously reported (30). In addition, microvascu-
lar embolism caused by the intracoronary administration
1995
of MSCs is related to cell number (13). Cheng et al. (6)
found that the intracoronary delivery of 1.0 × 106 CDCs
with or without magnetic targeting may result in microem-
bolization. Nakamae et al. (20) have found that an external
magnetic force can enhance adhesion of MSCs, which may
partly account for microembolization. How, then, can mag-
netically targeted intracoronary cell delivery resulting in
microvascular obstruction be improved? We speculate that
smaller cells may have the advantage of reducing embo-
lization. For example, very small embryonic-like stem
cells (~2–4 µm) found in recent years have been proven to
attenuate LV remodeling and improve LV systolic function
in small animal studies (8,33). In addition, previous study
found that sodium nitroprusside could influence the distri-
bution of MSCs by reducing the number of MSCs trapped
in the lungs (12). Vasodilator applications might represent
another promising method to reduce embolism.
The magnetic targeting is an emerging and promising
strategy in stem cell therapeutics. Before translation into
possible clinical application, it is warranted to elucidate
the optimized magnetic strength, exposure time, cell type,
cell number, and administration route. Because the infarct
size is larger and the ventricular wall is thicker in human
beings or large animals than those in rats, we speculate
that stronger magnetic strength is needed, and the vascu-
lar embolism may become a more serious complication.
Study Limitations
Although it describes a new and promising method, the
present study has a number of limitations. The optimized
magnet strength obtained from the small animal stud-
ies may not be directly extrapolatable to large animals.
Additionally, we did not include a magnet targeting group
without cell infusion and hence did not explore the effects
of magnet targeting on a rat model of acute ischemia/
reperfusion. Furthermore, instead of optimizing basal cell
dosage and magnetic exposure time, we chose cell dose
and duration consistent with previous studies that showed
functional benefits in small animals (5,6,9,18,19,25).
Further dosage and exposure time optimization would be
valuable, to inform future clinical studies. Finally, we did
not investigate whether the SPIO movement guided by
the magnetic field damaged the MSCs’ micro- and ultra-
structures. Future studies will be designed to capture any
changes caused by magnetic targeting in transplanted cell
micro- and ultrastructures.
CONCLUSION
We conclude that cell retention can be continuously
increased with the enhancement of magnetic targeting
intensity. Magnetic targeting can enhance cell engraft-
ment and improve therapeutic benefits through increasing
the effectively homing MSCs in a magnetic intensity-
dependent manner. However, when the magnetic field
1996
strength is elevated to a certain extent, microemboliza-
tion is increased, thus reducing long-term cell engraft-
ment and further undermining therapeutic benefit.
ACKNOWLEDGMENTS: The authors gratefully acknowl-
edge the financial support from the National Natural Science
Foundation of China (81370003, 81000043, 11304194, and
81100145) and the Science and Technology Commission of
Shanghai Municipality (13JC1401703). The authors declare no
conflicts of interest.
REFERENCES
1. Bayne, K. Revised Guide for the Care and Use of
Laboratory Animals available. American Physiological
Society. Physiologist 39:199, 208–211; 1996.
2. Boopathy, A. V.; Pendergrass, K. D.; Che, P. L.; Yoon, Y. S.;
Davis, M. E. Oxidative stress-induced Notch1 signaling pro-
motes cardiogenic gene expression in mesenchymal stem
cells. Stem Cell Res. Ther. 4:43; 2013.
3. Castro, E.; Mano, J. F. Magnetic force-based tissue engi-
neering and regenerative medicine. J. Biomed. Nanotechnol.
9:1129–1136; 2013.
4. Chaudeurge, A.; Wilhelm, C.; Chen-Tournoux, A.; Farahmand,
P.; Bellamy, V.; Autret, G.; Menager, C.; Hagege, A.;
Larghero, J.; Gazeau, F.; Clement, O.; Menasche, P. Can
magnetic targeting of magnetically labeled circulating cells
optimize intramyocardial cell retention? Cell Transplant.
21:679–691; 2012.
5. Cheng, K.; Li, T. S.; Malliaras, K.; Davis, D. R.; Zhang,
Y.; Marban, E. Magnetic targeting enhances engraftment
and functional benefit of iron-labeled cardiosphere-derived
cells in myocardial infarction. Circ. Res. 106:1570–1581;
2010.
6. Cheng, K.; Malliaras, K.; Li, T. S.; Sun, B.; Houde, C.;
Galang, G.; Smith, J.; Matsushita, N.; Marban, E. Magnetic
enhancement of cell retention, engraftment, and functional
benefit after intracoronary delivery of cardiac-derived stem
cells in a rat model of ischemia/reperfusion. Cell Transplant.
21:1121–1135; 2012.
7. Clifford, D. M.; Fisher, S. A.; Brunskill, S. J.; Doree, C.;
Mathur, A.; Clarke, M. J.; Watt, S. M.; Martin-Rendon, E.
Long-term effects of autologous bone marrow stem cell
treatment in acute myocardial infarction: Factors that may
influence outcomes. PLoS One 7:e37373; 2012.
8. Dawn, B.; Tiwari, S.; Kucia, M. J.; Zuba-Surma, E. K.;
Guo, Y.; Sanganalmath, S. K.; Abdel-Latif, A.; Hunt, G.;
Vincent, R. J.; Taher, H.; Reed, N. J.; Ratajczak, M. Z.;
Bolli, R. Transplantation of bone marrow-derived very
small embryonic-like stem cells attenuates left ventricular
dysfunction and remodeling after myocardial infarction.
Stem Cells 26:1646–1655; 2008.
9. Erwin, G. S.; Crisostomo, P. R.; Wang, Y.; Wang, M.;
Markel, T. A.; Guzman, M.; Sando, I. C.; Sharma, R.;
Meldrum, D. R. Estradiol-treated mesenchymal stem cells
improve myocardial recovery after ischemia. J. Surg. Res.
152:319–324; 2009.
10. Fukushima, S.; Varela-Carver, A.; Coppen, S. R.; Yamahara,
K.; Felkin, L. E.; Lee, J.; Barton, P. J.; Terracciano, C. M.;
Yacoub, M. H.; Suzuki, K. Direct intramyocardial but
not intracoronary injection of bone marrow cells induces
ventricular arrhythmias in a rat chronic ischemic heart fail-
ure model. Circulation 115:2254–2261; 2007.
11. Furlani, D.; Ugurlucan, M.; Ong, L.; Bieback, K.;
Pittermann, E.; Westien, I.; Wang, W.; Yerebakan, C.;
SHEN ET AL.
Li, W.; Gaebel, R.; Li, R. K.; Vollmar, B.; Steinhoff, G.;
Ma, N. Is the intravascular administration of mesenchymal
stem cells safe? Mesenchymal stem cells and intravital
microscopy. Microvasc. Res. 77:370–376; 2009.
12. Gao, J.; Dennis, J. E.; Muzic, R. F.; Lundberg, M.;
Caplan, A. I. The dynamic in vivo distribution of bone
marrow-derived mesenchymal stem cells after infusion.
Cells Tissues Organs 169:12–20; 2001.
13. Grieve, S. M.; Bhindi, R.; Seow, J.; Doyle, A.; Turner,
A. J.; Tomka, J.; Lay, W.; Gill, A.; Hunyor, S. N.; Figtree,
G. A. Microvascular obstruction by intracoronary delivery
of mesenchymal stem cells and quantification of resulting
myocardial infarction by cardiac magnetic resonance. Circ.
Heart Fail. 3:e5–e6; 2010.
14. Huang, Z.; Pei, N.; Wang, Y.; Xie, X.; Sun, A.; Shen, L.;
Zhang, S.; Liu, X.; Zou, Y.; Qian, J.; Ge, J. Deep magnetic
capture of magnetically loaded cells for spatially targeted
therapeutics. Biomaterials 31:2130–2140; 2010.
15. Huang, Z.; Shen, Y.; Pei, N.; Sun, A.; Xu, J.; Song, Y.;
Huang, G.; Sun, X.; Zhang, S.; Qin, Q.; Zhu, H.; Yang, S.;
Yang, X.; Zou, Y.; Qian, J.; Ge, J. The effect of nonuniform
magnetic targeting of intracoronary-delivering mesenchy-
mal stem cells on coronary embolisation. Biomaterials
34:9905–9916; 2013.
16. Huang, Z.; Shen, Y.; Sun, A.; Huang, G.; Zhu, H.; Huang, B.;
Xu, J.; Song, Y.; Pei, N.; Ma, J.; Yang, X.; Zou, Y.; Qian, J.;
Ge, J. Magnetic targeting enhances retrograde cell reten-
tion in a rat model of myocardial infarction. Stem Cell Res.
Ther. 4:149; 2013.
17. Meluzin, J.; Mayer, J.; Groch, L.; Janousek, S.; Hornacek, I.;
Hlinomaz, O.; Kala, P.; Panovsky, R.; Prasek, J.; Kaminek, M.;
Stanicek, J.; Klabusay, M.; Koristek, Z.; Navratil, M.; Dusek,
L.; Vinklarkova, J. Autologous transplantation of mononuclear
bone marrow cells in patients with acute myocardial infarc-
tion: The effect of the dose of transplanted cells on myocardial
function. Am. Heart J. 152:975.e9–975.e15; 2006.
18. Molina, E. J.; Palma, J.; Gupta, D.; Gaughan, J. P.; Houser,
S.; Macha, M. Right ventricular effects of intracoronary
delivery of mesenchymal stem cells (MSC) in an ani-
mal model of pressure overload heart failure. Biomed.
Pharmacother. 63:767–772; 2009.
19. Molina, E. J.; Palma, J.; Gupta, D.; Torres, D.; Gaughan,
J. P.; Houser, S.; Macha, M. Improvement in hemodynamic
performance, exercise capacity, inflammatory profile, and
left ventricular reverse remodeling after intracoronary deliv-
ery of mesenchymal stem cells in an experimental model
of pressure overload hypertrophy. J. Thorac. Cardiovasc.
Surg. 135:292–299; 2008.
20. Nakamae, T.; Adachi, N.; Kobayashi, T.; Nagata, Y.;
Nakasa, T.; Tanaka, N.; Ochi, M. The effect of an exter-
nal magnetic force on cell adhesion and proliferation of
magnetically labeled mesenchymal stem cells. Sports Med.
Arthrosc. Rehabil. Ther. Technol. 2:5; 2010.
21. Pfeffer, M. A.; Pfeffer, J. M.; Fishbein, M. C.; Fletcher,
P. J.; Spadaro, J.; Kloner, R. A.; Braunwald, E. Myocardial
infarct size and ventricular function in rats. Circ. Res.
44:503–512; 1979.
22. Pouzet, B.; Vilquin, J. T.; Hagege, A. A.; Scorsin, M.;
Messas, E.; Fiszman, M.; Schwartz, K.; Menasche, P.
Factors affecting functional outcome after autologous skel-
etal myoblast transplantation. Ann. Thorac. Surg. 71:844–
850; discussion 850–851; 2001.
23. Quyyumi, A. A.; Waller, E. K.; Murrow, J.; Esteves, F.; Galt,
J.; Oshinski, J.; Lerakis, S.; Sher, S.; Vaughan, D.; Perin, E.;
MAGNETIC INTENSITY FOR STEM CELL TARGETING
Willerson, J.; Kereiakes, D.; Gersh, B. J.; Gregory, D.;
Werner, A.; Moss, T.; Chan, W. S.; Preti, R.; Pecora, A. L.
CD34(+) cell infusion after ST elevation myocardial infarc-
tion is associated with improved perfusion and is dose
dependent. Am. Heart J. 161:98–105; 2011.
24. Robey, T. E.; Saiget, M. K.; Reinecke, H.; Murry, C. E.
Systems approaches to preventing transplanted cell death
in cardiac repair. J. Mol. Cell. Cardiol. 45:567–581; 2008.
25. Saito, T.; Kuang, J. Q.; Lin, C. C.; Chiu, R. C. Transcoronary
implantation of bone marrow stromal cells ameliorates
cardiac function after myocardial infarction. J. Thorac.
Cardiovasc. Surg. 126:114–123; 2003.
26. Sanganalmath, S. K.; Bolli, R. Cell therapy for heart fail-
ure: A comprehensive overview of experimental and clini-
cal studies, current challenges, and future directions. Circ.
Res. 113:810–834; 2013.
27. Tambara, K.; Sakakibara, Y.; Sakaguchi, G.; Lu, F.;
Premaratne, G. U.; Lin, X.; Nishimura, K.; Komeda, M.
Transplanted skeletal myoblasts can fully replace the
infarcted myocardium when they survive in the host in large
numbers. Circulation 108(Suppl 1):II259–II263; 2003.
28. Terrovitis, J. V.; Smith, R. R.; Marban, E. Assessment and
optimization of cell engraftment after transplantation into
the heart. Circ. Res. 106:479–494; 2010.
29. Toma, C.; Wagner, W. R.; Bowry, S.; Schwartz, A.;
Villanueva, F. Fate of culture-expanded mesenchymal stem
cells in the microvasculature: In vivo observations of cell
kinetics. Circ. Res. 104:398–402; 2009.
30. Vulliet, P. R.; Greeley, M.; Halloran, S. M.; MacDonald,
K. A.; Kittleson, M. D. Intra-coronary arterial injection of
mesenchymal stromal cells and microinfarction in dogs.
Lancet 363:783–784; 2004.
1997
31. Wang, X.; Jameel, M. N.; Li, Q.; Mansoor, A.; Qiang, X.;
Swingen, C.; Panetta, C.; Zhang, J. Stem cells for myocar-
dial repair with use of a transarterial catheter. Circulation
120:S238–S246; 2009.
32. Wohrle, J.; von Scheidt, F.; Schauwecker, P.; Wiesneth, M.;
Markovic, S.; Schrezenmeier, H.; Hombach, V.; Rottbauer,
W.; Bernhardt, P. Impact of cell number and microvascular
obstruction in patients with bone-marrow derived cell ther-
apy: Final results from the randomized, double-blind, pla-
cebo controlled intracoronary stem cell therapy in patients
with acute myocardial infarction (SCAMI) trial. Clin. Res.
Cardiol. 102:765–770; 2013.
33. Wu, J. H.; Wang, H. J.; Tan, Y. Z.; Li, Z. H. Characterization
of rat very small embryonic-like stem cells and cardiac
repair after cell transplantation for myocardial infarction.
Stem Cells Dev. 21:1367–1379; 2012.
34. Xu, J.; Qian, J.; Xie, X.; Lin, L.; Zou, Y.; Fu, M.; Huang, Z.;
Zhang, G.; Su, Y.; Ge, J. High density lipoprotein protects
mesenchymal stem cells from oxidative stress-induced
apoptosis via activation of the PI3K/Akt pathway and
suppression of reactive oxygen species. Int. J. Mol. Sci.
13:17104–17120; 2012.
35. Yao, K.; Huang, R.; Sun, A.; Qian, J.; Liu, X.; Ge, L.;
Zhang, Y.; Zhang, S.; Niu, Y.; Wang, Q.; Zou, Y.; Ge, J.
Repeated autologous bone marrow mononuclear cell ther-
apy in patients with large myocardial infarction. Eur. J.
Heart Fail. 11:691–698; 2009.
36. Zhang, C.; Liu, T.; Su, Y.; Luo, S.; Zhu, Y.; Tan, X.; Fan,
S.; Zhang, L.; Zhou, Y.; Cheng, T.; Shi, C. A near-infrared
fluorescent heptamethine indocyanine dye with preferen-
tial tumor accumulation for in vivo imaging. Biomaterials
31:6612–6617; 2010.