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Behind the Scenes of Cell Culture

Laboratories


by Carlo Jochems, BSc, Candidate MSc Molecular Sciences,

Wageningen University, The Netherlands


    Cell culture techniques represent a way of performing scientific, medical and toxicity research without the use of live animals.  At least, this is the view held by most people, but is it true? NO!

for two reasons.


    First, several animals have to be killed for the purpose of obtaining cells.  For example, in 1993, the number of animals killed in France to  obtain their cells was 111,792; in 1995; the number killed in The Netherlands was 114,012.  Luckily, cell lines can grow in a plastic flask for many more generations  than in the original animal so that the net result is that fewer live animals are used.


    Second,  a suitable growing medium is required to culture the cells.  In many cases this involves the use of fetal bovine serum (FBS) which is obtained from fetal calves.  Serum is blood without cells and clotting factors. 


   Most of it comes from countries where cows are kept in an extensive manner as in Australia and some American countries where cows and bulls are allowed to roam together.  The bulls serve to keep the herd together, but there is a side effect.  Some of the cows will be pregnant when the herd is sent for slaughter.  This is common practice.  Whether or not a cow is pregnant is simply not an issue!


    If the slaughterhouse has the right facilities, the entire reproductive tract including the fetus is removed and taken to the "calf processing area" right after a pregnant cow is killed. There, the fetus is cut out of the uterus.  The umbilical cord is tied off.  The amniotic fluid is cleaned off and the fetus is disinfected.


    Then, a large diameter needle is inserted through the skin and between the ribs, directly into the beating heart of the unanesthetized  fetus.  The blood is commonly extracted by vacuum into a sterile collection bag.

 

    The blood is then allowed to clot at a low temperature and the clotted substance separated from the serum by refridgerated centrifugation.  The fetus is then destroyed to assure it will not be used for human consumption.


    "For all practical purposes, a fetal calf's heart must be beating to obtain an adequate harvest for fetal calf serum production," according to Gene Erickson, a former US Department of Agriculture  inspector.  Several producers and authorities in countries where FBS is extracted deny the fetus is alive, however.  "...the fetus is dead at the time of cardiac puncture" claims K. Johar of the Australian Quarantine and Inspection Service, Technical Services Branch.


    New Zealand Ministry of Agriculture and Fisheries Animal Welfare Advisory Committee guidelines for the welfare of livestock used for blood collection explain (p.6) that the cardiac puncture "...occurs about 20 to 30 minutes after the cow has been killed and the fetus will have been dead for almost as long. These are post mortem procedures which do not require Animal Ethics Committee approval."


    The same guidelines, however, state on page 14: "Effective electrical stunning causes cardiac arrest and so electrical stunning is not suitable for the harvesting of blood from the heart."  When

I later asked Birgit Petroschka of Kraeber GmbH, a German FBS supplier, to comment on these contradictory statements, the answer was: "It is absolutely impossible to collect blood from fetuses which are dead for 20 - 30 minutes.  I asked one of the big FBS suppliers in New Zealand and they stated to me that FBS is produced in a normal way, that means by heart puncture."


    As John Walsh of the World Society for the Protection of Animals (WSPA) put it: "...there is a great deal of cover up and government ignorance as to the actual sources of neo-natal and fetal serum.  Please keep in mind that the federal and municipal ministries of agriculture work closely with such industries and in most cases protect such industries...The industry is secretive and cautious."


    Not all FBS suppliers cover up.  "The animal is alive when bleeding," comments  Chilean producer A. Cox of Comercial e Industrial APREX Ltda..  Petroschka adds: "The blood has to be taken by cardiac puncture from the living fetus.  When an animal dies, the blood is coagulating immediately...The heart must pump to transport the liquid blood outside the body."


    The heart functions. The fetus is alive.  But the fetus is not receiving any form of anesthesia prior to being subjected to cardiac puncture, an extremely painful procedure.  Over the last 10 to 15 years, more and more scientific data has been piling up showing that fetuses of mammals (particularly those species whose newborns are relatively well-developed at birth like cows, horses, guinea pigs, sheep, goats and pigs) can experience pain or discomfort well before birth.  A recent guideline on humane euthanasia of experimental animals reported that fetuses could experience pain or discomfort after 30% gestation time.  Bovine fetuses are not usable for extracting FBS at less than three months gestation, which is 33% of normal gestation. 


Commonly, they have experienced six months or more gestation.  So all the fetuses used are capable of experiencing pain but are never anesthetized.


    Worse yet, mammal fetuses from a certain time in their development are even more susceptible to pain than adults! It is argued that lack of oxygen after the mother cow is killed may cause some degree of unconsciousness, but mammal fetuses are highly resistant to oxygen deprivation and the degree of unconsciousness at the moment of puncture has never been scientifically investigated.


    Ethical scientists would err on the side of caution wherever animal suffering is concerned not on the side of ignorance or disinterest.  This is apart from the question of whether man has
the right to use animals for human interests.


    Says Petroschka: "I know that (this) is an ethical problem, but the fetal bovine serum is an essential product for cell culture in science, clinical health care and biotechnology."


    Michelle Howald of the Animal Welfare Division of the Swiss Ministry of Veterinary Affairs comments: "At least the use of FBS is unethical whenever it is used for habitual reasons in tests where one could do without orreplace FBS by synthetic medium."


    According to R. Watt of the UK Animals (Scientific Procedures)
Inspectorate, the procedure would be considered an animal experiment.


    Petroschka further states: "Countries like Hungary, the former Russian-Baltic countries and maybe Czechoslovakia have a commercial production of FBS, that means they are breeding fetal cows for the production of FBS. This is the reason we do not buy raw material from such countries."


    Apparently, there is something of a
contradictio in vitro when it comes to the use of FBS in culturing cells as an alternative to animal experiments. 


    Around 500,000 liters of FBS are obtained from over a million bovine fetuses worldwide. The best alternative is to use 100% chemically manufactured media. The overall conclusion is that cell culture work does not free us from animal welfare considerations as taken for granted by many!

------------------------


For more information on FBS and its alternatives, see http://prex.las.vet.uu.nl/nca/ . Select <Documents>, which contains the summary of a report on livestock blood sera production methods and related ethical implications as well as a downloadable version of the report with many references.


Alternatives to Laboratory Animals has accepted an article on this issue which will probably be published before the end of the year. The Netherlands Centre (for) Alternatives (to animal use) is considering a workshop in cooperation with the European Centre for the Validation of Alternative Methods. As this is the first time this issue has been touched on in a scientific manner, a workshop would be the proper vehicle for arriving at conclusions*** regarding the animal welfare aspects of this form of unborn animal use, if not abuse.   - C. Jochems


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The Civil Abolitionist

Autumn 2000

The Civil Abolitionist

Autumn 2000