In Vitro Meat Production

Course: Management Information Systems

Instructor: Prithwis Mukerjee
Nachiketa Trivedi
Vinod Gupta School Of Management, IIT Kharagpur

Abstract

In this paper we discuss the concept of producing edible meat in vitro by culturing animal meat in
laboratory or at a commercial scale. The production of highly structured skeletal muscle tissue is a
great technical challenge. In the short run this method of meat production may remain economically
infeasible. But ongoing research and economies of scale may make it the alternative that we are
looking for to feed the growing meat and meat product demand in the developed as well as
developing world. It has the potential to pertinent questions like food security, rising meat
consumption in the developing world, global warming and food borne infections and the lifestyle
diseases arising out of increased meat consumption. We would explore the methods of production of
in vitro meat production its advantages, disadvantages and its implication at large.

What is In Vitro meat production?

It is feasible to grow cells of plants and animals in the laboratory without the organism from which the
cell was derived. This practice is known as in vitro culturing. Culturing involves extraction of cells from
an organism and growing the cells in laboratory by using the appropriate culture medium under
controlled conditions of temperature, pH and other factors. These cells can be grown for various
purposes like research, medicine production etc. In this paper we would look at technique used to
grow cells or rather meat in the laboratory. This presents a unique challenge of growing a large
number of cells from a small number of extracted cells and also to grow them as skeletal muscle
tissues or edible meat.

But before looking at the processes involved in actually growing the cells in the laboratory and
producing edible meat commercially let us look at why meat consumption is necessary and what
factors are driving us to explore the possibility of In Vitro meat production.

Why is Meat Consumption Essential?

Requirement of Human body for animal protein

Human body has evolved as omnivore. To maintain good health a balanced diet containing both
vegetarian and non-vegetarian components is needed. Although the vegetarian diet contains almost
all the nutrients needed by the body but is completely devoid of certain components like vitamin B12.
Vitamin B12 (cobalamin) is required by bacteria, algae and animals, but not by higher plants.
Therefore, it is completely absent in food sources derived from higher plants. All vitamin B12 is
ultimately derived from bacteria and algae, which are all but a few exceptions able to synthesize it. It
enters the food chain of higher animals via large herbivores, where it is produced by the in the gastro-
internal tract. This is not enough to fulfil the requirements, and even if it did it could not be absorbed
by their intestine. A lack of vitamin B12 leads to the conditions of megaloblastic anaemia1 and sub
acute combined degeneration2, and is lethal in the long. Hence a vegan diet is probably not possible
without the intake of (chemically derived) food additions. Hence there is a genuine need for non
vegetarian component in a balanced diet.
It is possible to speculate about two other reasons why the consumption of animal products is desired
over a diet lacking animal products altogether. The first is that man has evolved as an omnivore, so its
gastro-internal regulation and its physiology are optimized for the processing of an omnivorous diet.
This could imply that some of the essential nutrients are digested best when it is consumed in form of
omnivorous diet. There also one section of scientists who believe that the present research on dietary
requirements of human body is still not comprehensive and we would better understand the true
importance of an omnivorous diet as researchers probe deeper into the mystery of diet, digestion and
requirements of the body.

Trends in Meat Consumption

Apart from the nutritional status, meat and other animal products play an important social role in the
western society. In The Netherlands, the per capita meat consumption was 50.1 kg 2001. In the
period 1994/1995 this amount was for Germany and Switzerland respectively 61.6 and 57.7 kg. A
survey in Switzerland showed that 24.8% of the respondents consumed meat or meat products at
least once every day. The consumption of meat appears to be a strong social factor, as only a
marginal group chooses to be vegetarian; 4.7% of the adults in Australia in 1995, 5.4% of the adults in
Britain in 1997. From 1961 on there is a trend in both developed countries as in developing countries
to eat more meat. Until 2020, meat demand is expected to increase highly in developing countries
and slightly in developed counties.

Poor people everywhere are eating more animal products as their incomes rise above poverty level
and as they become urbanized. By 2020 developing countries would consume 63% of the world’s total
meat as against 52% at present this would translate to an increase of 107million metric tonnes
dwarfing the increase of 19 million metric tonnes in the developing countries. The projected increase
in livestock production will require annual feed consumption of cereals to rise by nearly 300 million
metric tonnes by 2020. This increased demand is driving a Livestock Revolution.
 It is imperative to look at the aspects of this livestock revolution. This would help in understanding
environmentally and economically unsustainably nature of the increasing meat consumption.
Important global livestock sector trends reflecting this ‘revolution’ are:

a rapid increase in consumption of livestock products in developing as well as developed countries

Shift in the geographical area of livestock farming from cold and dry area to humid wet and warmer
area thus rendering the industry more prone to animal disease break outs.

The change in the nature of business from a non commercial or small scale commercial activity to a
organized large scale commercial activity

And most importantly basing of increasing proportions of animal feed on cereals.
Thus meat consumption is bound to grow with the increase in demand of meat products in developed
as well as developing countries. The rising consumption would put pressure on resources and would
also have environmental and economic consequences.

Problems associated with meat production and Consumption

Although meat has enjoyed sustained popularity as a foodstuff, in recent years consumers have
expressed growing concern over some consequences of meat consumption and production. These
include:

Health Problems: Overconsumption of meat has an adverse on the body. It is also responsible for the
rising instances of diseases such as obesity heart diseases and constipation. Consumption of large
quantities of meat is also linked to cancer. These diseases are now responsible for a third of global
mortality. A survey conducted in California in 1960’s found that the risk of heart disease is three times
greater for 45-64 year old men who eat meat daily, versus those who did not eat meat. In another
study in 2010 involving over one million people who ate meat found that processed meat had an
adverse risk in relation to coronary heart disease. The study found that having 50g of processed meat
per day increases risk of coronary heart disease by 42% and diabetes by 19%. These adverse effects
of meat consumption pose a genuine health threat which needs to be addressed.

Food Borne Diseases: Another problem associated with consumption of meat is the issue of food
borne diseases. Pathogens in meat such as Salmonella, Campylobacter, pathogenic E. coli, avian
influenza, cause outbreaks of epidemics which effect the meat consuming population at large. This
has been proved by the outbreak of bovine spongiform encephalopathy (BSE) in the UK and Europe
and the chicken flu in The Netherlands and Belgium. These diseases can be potentially dangerous for
humans. Consumption of certain types of beef affected by BSE can cause a rare disease called as
the Creutzfeld-Jacob disease. But in more recent times greater threat has been posed by the
outbreak of avian and H1N1 outbreak which assumed pandemic proportions within short time and
caused health and economic damage. In the United States, food-borne diseases – the most common
causes of which are contaminated meats -- are responsible for over 76 million episodes of illness,
325,000 hospitalizations, and 5,000 deaths each year.

Inefficient use of resources: Raising animals for meat requires considerably resource in form of more
cropland, water, fertilizer, pesticides, and energy. A lot of resources are consumed in maintaining
livestock rearing facilities, transporting livestock , growing animal feed, transporting animal feed,
maintaining slaughter houses and transporting meat thus intensive meat production is only 25
percent as energy efficient as soybean production. If we feed the burgeoning global population on
plant derived products then we can feed a significantly larger number of people.

Pollution: Annually, 1.4 billion tonnes of animal wastes are produced in the US. Cattle and sheep are
the second largest emitters of greenhouse gases that contribute to global warming, next to carbon
dioxide. This is because commercial animal farming techniques over-feed the animals which causes
them to produce more of these harmful gases than they normally do. Animals are the primary
producers of methane which is much more potent than carbon dioxide as far as green house potential
is concerned.Meat production is responsible for the emissions of nitrogen and phosphorus, pesticide
contamination of water, heavy metal contamination of soil, and acid rain from ammonia emissions.

Animal Suffering: Because of increasing commercialization of animal farming large number animals
are herded in closed spaces. This leads to high levels of stress in animals and even leads to
premature deaths. Pigs herded for meat consumption with intensive farming techniques live in small
concrete boxes. Food provides the only distraction, but this is consumed quickly due to its high
nutritional quality. This leads to such high growth rates which are unsustainable leading to diseases in
animals. Also, antibiotics are increasingly being used as growth promoters for animals kept in
intensive farming. This has been linked to the emergence of multi-drug-resistant strains of pathogenic
bacteria.

Due to the aforementioned factors there has been a continuous search for an alternative. In Vitro
Meat production technology is an alternative which can address all the above issues. As mentioned
before, In Vitro meat production refers to the practice of artificially growing animal or plant tissue in
laboratory. Before going into the details of the production technology let us look at structure,
development, and growth of skeletal muscles in vertebrates.

Anatomy of Skeletal Muscles

Skeletal muscles are composed of columns of muscle tissue called fascicles which are embedded in
fibrous connective tissue called as epimisium. The fascicles are in turn made up of mayofibres which
are elongated multinucleated cells. The muscles are classified into two groups called as the fast
muscles and the slow muscles. The fast muscles are specialized in reflexive fast actions and the slow
muscles are specialized in slow and prolonged activity.
Formation process of Skeletal Muscles

Cells need to withdraw from the natural cell division cycle to be able to form muscles. After the cells
withdraw from the regular cellular cycle, Muscle Regulatory Factor 4 (mrf4) and Myogenin (myogenin)
initiate muscle differentiation. It is proposed that there are two phases of proliferation of mayoblasts
the first one giving rise to slow muscles and the second one to fast muscles. Very little muscle cell
proliferation takes place in higher animals after embryogenesis. In postnatal muscle tissue, full-grown
muscle fibres are accompanied with satellite cells. These cells are the primary source for new nuclei
in postnatal muscle tissue. Satellite cells are distinct in their appearance. They initiate muscle growth
by donating their DNA to existing muscle fibres or fusing themselves to form muscle fibres. Normally,
satellite cells are quiescent and non-proliferative, but upon stimulation they start to proliferate.
The objective is to isolate the satellite cells which have withdrawn from the cell cycle and stimulate
their proliferation. Embryonic stem cells can also be used for the purpose but they haven’t been found
as suitable as satellite cells the reasons for which we would explore in the coming sections. Typically
these satellite cells are more abundant in the muscle cells of neonatal individuals than in the muscles
of older individuals. Abundance is not the only criteria for the selection of individuals another factor is
the proliferative capacity of the cells of the individual. This capacity is negatively related to the age of
the individual. The fall in the proliferative capacity may not be proportional with age. The satellite cells
of a human infant are capable of multiplying 55–65 times, but at the age of 9 this number is about 45
divisions. In addition, older satellite cells form thinner and more fragile mayotubes. Isolation requires
mincing the muscles, followed by enzymatic treatment and subsequent separation of the satellite cells
by differential centrifugation.

Cultured Meat Production

Skeletal muscle fibres constitute edible animal meat. The production of cultured meat in vitro thus
requires techniques of skeletal tissue engineering. These techniques can be divided into two
categories scaffold based techniques and self organizing techniques.
In scaffold based techniques scaffolds or carriers are used as the basic structures upon which the
satellite cells grow. A scaffold or carrier can be a collagen meshwork or micro-carrier beads. The
mesh work is seeded with cells. The meshwork seeded with cells is then perfused with the
appropriate culture medium in a bioreactor or the culture medium is percolated through the
meshwork. Various environmental cues are provided which stimulates the cells to fuse and thus form
myotubes which differentiate into myofibers. The mayofibers obtained are then harvested as meat and
can be consumed.
A scaffold based technique is useful for producing meat to be consumed as hamburgers or minced
meat but for meat preparation such as steaks where the structure of the meat is important scaffold
based techniques don’t serve the purpose. For such structured meat pieces it is necessary to grow
meat as self organizing units.
To grow meat as self organizing constructs muscle explants have been grown with different culture
mediums. Explants have the advantage of containing all the cells that make up muscle in their
corresponding proportions. The growth has been dependent on the medium used . Although the
results of the experiments have been encouraging but the success has been limited due to the
absence of system to replicate the blood flow mechanism in actual muscle tissue. The proliferated
muscles die in case separated from the nutrient medium by more than .5mm.
Vascularisation is proving to be the crucial hurdle which is limiting the success of the efforts to grow
self organizing meat constructs in the laboratory. Proposed solutions suggest making a branched
network of edible polymer through which nutrients are circulated. The method involves creation of a
cast onto which a biochemical solution or a collagen is spread. After the polymers set the original
material is dissolved leaving behind a network of micro channels. Many such layers can be stacked
onto one another to give a three dimensional capillary network which can be used to mimic
vascularisation artificially.

Process Diagram

Source: In vitro cultured meat production P.D. Edelman, D.C. McFarland, V.A. Mironov, and J.G.
Matheny.

Scaffold-based cultured meat production:

1. Isolated Myoblasts
2. Porous collagen microspheres;
3. Myoblasts form myotubes on collagen microspheres;
4. Bioreactor;
5. Microwave;
6. Hamburger
Now we would look at the crucial aspects of in vitro production process in details and understand the
impediments and the challenges that lie ahead in implementing this technology on a mass scale.

What Type of Cells to Choose

Skeletal muscle is made up of several cell types. The proliferation differentiation and fusion of the
embryonic mayoblasts form skeletal muscle tissue. In post natal stage this is done through the
satellite cells. There is an option of using embryonic stem cells for in vitro meat production stem
systems. These cells have high potential to differentiate and proliferate. But the harvests from these
cells don’t justify the effort required to force them to differentiate. Thus satellite cells are the most
appropriate option for growing muscle cells in vitro. Satellite cells with high proliferative capacity have
been isolated from skeletal muscle of chickens, turkeys, pigs, lambs and cattle. These cells have
been grown in different medium to encouraging results. These cells can be harvested as meat for
consumption. Therefore, it is possible to isolate cells from animals grow them in a nutrient medium
and consume as meat. But for in vitro meat to taste just as natural meat would require a vascular
system and a connective tissue system organized in three dimensions.

Mayotubes grown from satellite cells of a turkey.

Source: In vitro cultured meat production P.D. Edelman, D.C. McFarland, V.A. Mironov, and J.G.
Matheny.

How Much Meat can be got through a single Cell

It has not been established that how much meat could a single cell yield in culture. Cells in culture
have a limited life span and undergo a fixed number of doublings in that life span. The number of
doublings that a cell could undergo in a lifespan is known as the Hayflicks limit. Cells with high
Hayflicks Limit could yield very high yields. Although the Hayflick limits of most of the domestic animal
cells have not yet been established it believed that cells of turkey could produce large quantity of
meat. It is estimated that cell with a Hayflick s Limit of 75 could satisfy the whole global demand of
meat. Even though some of the animal cells may not have high Hayflick Limits genetic engineering
offers a way out by modifying the telomerase structures. Thus cells with low Hayflick Limit can be
engineered to have higher yields.

Fields

The need for stimulation or exercise has been established for tissue culture systems. Atrophy is a
clear threat for an in vitro meat production system as there is a danger of growth retardation. In fact
muscle fibres formed in culture start to contract spontaneously in laboratory conditions. In line with
this is the fact that mechanical, electromagnetic, gravitational, and fluid flow fields could have a
significant impact on the differentiation of cells. It has been found that repetitive stretch and relaxation
equal to 10 percent of length, 6 times per hour, increased differentiation into myotubes. Differentiation
has been observed in myoblasts seeded with magnetic micro particles placed in a magnetic field,
without adding special growth factors or any conditioned medium. Electrical stimulation also
contributes to differentiation.

Scaffolds

For proliferation and differentiation to occur it is necessary that a substratum or scaffolding be

provided. This substratum or the scaffolding should be edible. As has been already discussed
mayotubes formed start stretching and contracting spontaneously and also that stretching and
contracting act as stimulants for growth. Therefore flexible scaffolding is necessary to encourage
growth and also to prevent mayofibres from detaching. Cytodex-3 microcarriers have been used as
scaffoldings but they don’t have any stretching potential and thus they are inappropriate. A more
appropriate option is to use a material which is sensitive to environmental cues and expands or
contracts as they are changed. An example is a material which would change in surface area with
change in temperature or pH.

Culture Media and Growth Factors

To be an economically feasible alternative to conventional meat production it is very important that the
medium used is non expensive. The medium should also be able to provide the necessary nutrients
to the mayoblast in simple form as no digestion is involved. There have been many developments on
this front. It is critical that the costs of the culture medium are reduced. It is so because cost of culture
medium is expected to be the biggest cost contributor in case of commercial scale in vitro meat
producing unit. The cost contribution is expected to be 1/10 of the operation cost. Recent
developments in which serum free media were used would certainly reduce costs of the culture
medium.
Bioreactors
 Meat production would require development of new bioreactors. These bioreactors should have low
shear and should maintain high perfusion at large volumes. Most skeletal muscle tissue research has
been done using the NASA rotating bioreactors. This is because the cells in the reactor remain in
continuous suspension, fluid shear in minimum and cell assemblies up to 1 cm can remain in
8
suspension. Bioreactors such as these are capable of sustaining cell concentrations of up to 10 cells
per mL. Bioreactors as small as 250 ml have been scales upto 3 litres and further expansion to
industrial scale would theoretically not affect the system. In fact as the reactors are scaled up the cost
of production would drop down and this would make large scale production feasible.

Uses and Advantages of Using In Vitro Meat Production

Health Benefits: As has been discussed consumption of meat has become the primary cause of
increasing instances of cardiovascular diseases, diabetes, obesity and even cancer. In Vitro meat can
be used to address this issue. It is high content of saturated fat in meat which is responsible for health
risks associated with meat consumption. In in-vitro meat the fat content can controlled by introducing
fat after the meat has been harvested or by co-culturing with adiposities. The fat percentage of
unsaturated fatty acids can be enhanced to have beneficial effect on blood cholesterol.

Disease control: The number of cases of food borne diseases could be significantly reduced if not
totally eliminated. This is because of the greater control over the manufacturing process. With
adoption of standard good practises and stringent quality measures which are very difficult to impose
on the long supply chain of meat production at present meat quality can be enhanced so as not to
cause any food borne diseases. Also as the meat production process would be carried out in large
factories there would not be any exposure to pesticides, arsenic, antibiotics and artificial hormones.

Efficient use of Resources: When animals are raised in a farm for meat, energy is also needed for the
metabolic functions of the animals like maintaining body temperature, reproduction etc. The animal
has to be developed fully in terms of all the body parts like brain, skin etc. But in case of in vitro meat
only parts of animals which have to be consumed have to be grown and also no energy would be
consumed in the metabolic and locomotion functions of the animals. This would lead to very efficient
use of resources.

Less Time Consuming: In Vitro meat production would lead to less time being consumed for meat
production. Whereas conventional meat production takes years to raise livestock and harvest meat in
vitro meat production system would only take a few weeks to produce edible meat.

In Vitro meat production would minimize waste production from the meat production process.

As livestock would not have to be maintained and in vitro meat would be produced in industrial units
therefore the process would free up a lot of land that has been engaged in the activity of livestock
 maintenance. This would present a great economic opportunity to use this land for purposes such as
housing, plantations and also forest regeneration. This effect would be more pronounced in countries
like Argentina and Brazil which have large proportion of land being used for meat production process.
This can address the problem of additional crop land needed and land for affordable housing in these
countries.

Species which are on the brink of extinction due to poaching for meat can be saved by producing their
meat in the laboratory.

The concept of production of meat in vitro is not limited to producing muscle tissue for human
consumption. The concept can be extended to tissue restoration and regeneration. It can be used to
grow human muscles and then help in healing. This link between the in vitro technology and
regenerative medicine technology can synergise developments in both fields. Any development in the
in vitro meat technology can be a boost for the regenerative technology and vice versa.

In Vitro meat also finds a place in the bio-regenerative systems as a means of providing food for
astronauts on long haul space missions. Actually carrying food conventionally represents a logistical
problem for space travel as costs of launching payload is astronomical. Therefore a bio-regenerative
system was envisaged which would recycle human waste to build a self sustaining system. The
question is why would an in vitro meat would be a essential part of such a system. There two reasons
for this first that the efficient digesters of human waste like fungi and algae are not always edible by
humans. Secondly, plant matter if generated in the recycling process would lead to waste as two
thirds of plant matter is not digestible by humans. If the plant matter is decomposed it would again
lead to the formation of fungi or algae. But the fungi and algae formed in the process act as good
substrate for the formation in vitro meat. Thus in vitro meat would be an essential part of a
regenerative system. In fact this use is not limited to space technology. It can be used where the cost
of regeneration of human waste is less than the cost of transporting the food for example polar-
regions, remote defence posts or bunkers.

In vitro meat would also lead to reduced animal use. Theoretically only a few animals can be used to
source the entire meat requirement.

Consumer Acceptability

To be implemented as large scale commercial meat production system the in vitro meat would have
to be accepted by consumers. For in vitro meat to be acceptable by the customers it is necessary that
the taste and texture closely resemble the natural meat. The fact that meat would not exactly
resemble natural meat is offset by the fact that there is large market for boneless and skin less meat
and use of meat for processed meat products such as hamburgers and sausages. It would also
appeal to increasing number of people concerned about food security and also to those who are
 against killing of animals for meat. However there is an intuitive reluctance to reject synthesised
foods. It is this factor which could lead to non acceptance of in vitro meat.

Suggestions to make in vitro meat more acceptable

Raise customer awareness about the environmentally unsustainable nature of meat production today

Underline the benefits to the environment and the economic benefits that would accrue from the
usage of in vitro meat

Stress on the health advantages of having in vitro meat as has been discussed in the paper

Stress should be laid on the fact that use of in vitro meat would reduce the atrocities being committed
on animals, like overfeeding, confining in small spaces and use of antibiotics and hormones to
accelerate growth, in the name of commercial production of meat.

Pertinent question of food security can also be used to sway public opinion in favour of in vitro meat.

Economy

At present the production the commercial production of in vitro meat is not economically possible. It is
estimated that at present it would cost $5million to produce one kilogram of functional skeletal muscle
in laboratory.

Financial studies conducted into the feasibility of in vitro meat production reveals a different picture. At
present the studies conducted assume that the upfront R&D costs would be huge and would be
funded by governments or through research funding. The capital costs have been estimated on the
basis of currently available technologies although the exact technologies to be used for in vitro meat
production are not known at present and could be cheaper. Medium costs would be the major costs
for the use of the technology they have been estimated on the basis of available media costs and
have been discounted for mass production. Proceeding with the assumption of mild financial risks,
estimates suggest that in vitro meat can be available at 3300-3500 euro/tonne. This cost is
comparable to the price of unsubsidized chicken production of 1800 euro/tonne. These estimates
clearly suggest that this technology is not unfeasible and requires certain changes which can reduce
costs.

Proposals to Reduce Costs

Costs would certainly be reduced if suspended cell culture technique is used and the meat is
produced in industrial scale bioreactors.

The economies of scale would certainly reduce costs.

It would be advantageous if the units producing the culture media and the production units are located
at the same site.

Concept of using fungus or algae based media derived from decomposition of human or plant waste
can be investigated as this can drastically reduce costs.

Advances in cell culture technology would further improve the efficiency of the process. Theoretically
in vitro meat production would give higher labour and resource efficiency and thus could be cheaper
than conventional meat if it is grown in an affordable culture medium.

Conclusion

In vitro meat production method is a technology of the future. It has been proved feasible technically
but the commercial implementation of the technology of the process would require better
understanding of the processes involved like scaffold technology, environment cues and their effect
on cell culture and the culture technology. The most crucial aspect that would need to be addressed is
the culture medium technology. Subsequently measures should be taken to enhance customer
acceptability of the in vitro meat. In vitro meat production technology has the potential to address the
growing concerns of humanity like food security and global warming and at the same time as has
been discussed in the paper that it is feasible at reasonable cost. With advance in technology these
costs would only reduce. In view of the advantages and the feasibility it is imperative that in vitro meat
technology be encouraged.

References

[1] The In Vitro Meat Consortium Preliminary Economics Study Project 29071 V5 March 2008.

[2] In Vitro Meat: Zombies On The Menu? Neil Stephens

[3] In vitro meat production Pieter D. Edelman 17th November 2003.

[4] In vitro cultured meat production P.D. Edelman, D.C. McFarland, V.A. Mironov, and J.G.
Matheny.