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A Severe Case of Fraudulent Blending of Fetal Bovine Serum Strengthens the Case for Serum-free Cell and Tissue Culture Applications

Authors:
In 2011, GE Healthcare (a unit of General Electric
Co.) acquired PAA Laboratories, Linz, Austria. In
April 2013, GE Healthcare published a product
information to customers, stating that batches of
fetal bovine serum (FBS) produced at PAA facili-
ties from March 2008 to March 2013 are subject to
label non-conformances, i.e. that:
“These products may contain added adult bovine
serum albumin (BSA) of United States origin,
water, and/or cell growth promoting additives. For
FBS product shipped into countries other than the
United States, current product labeling states that
the origin of the product is either Australia or EU
approved serum sources. In addition to, or instead
of product of this origin, the product may contain
adult BSA of United States origin and/or may
contain FBS from sources including United States,
Canada, Argentina, Brazil, and/or Mexico.”
This warning of GE Healthcare about the purity
and quality of FBS from PAA Laboratories
prompted us to write a note to inform and to alert
the cell culture community, and to provide back-
ground information about FBS, and about serum
alternatives and serum-free cell culture applica -
tions, respectively.
1
FBS is a natural cocktail of most of the factors
required for cell attachment, growth and prolifera-
tion, effective for most types of human and animal
(including insect) cells.
2
Although in use as a
universal growth supplement of cell and tissue
culture media for more than 50 years, FBS has
never been fully characterised. Recent proteomic
and metabolomic studies revealed approximately
1,800 different proteins
3,4
and more than 4,000
metabolites present in serum,
5
with the proportions
of each of these components varying between
different serum batches. Furthermore, global
supply and availability of FBS has changed dramat-
ically over the past few years. FBS is a by-product of
the beef packing industry. Thus, FBS supply is
dictated by many factors, including beef consump-
tion (e.g. more white meat over red meat), feed
prices, environmental factors such as drought,
cattle import and export, governmental farm poli-
cies,
6
and the outbreak of diseases (e.g. foot and
mouth disease, BSE).
7–10
From this, it can be concluded that the use of
serum in cell culture may involve a number of
disadvantages: a) serum in general is an ill-defined
supplement in culture media, with high qualitative
and quantitative, geographical and seasonal batch-
to-batch variations; b) FBS may contain adverse
factors, like endotoxins, mycoplasma, viral
contaminants or prion proteins; c) there are animal
welfare concerns surrounding the harvest and
collection of FBS from unborn bovine fetuses; and
d) FBS availability is dependent on the global
market.
11–13
There is a severe geographical mismatch between
the supply of, and the demand for, FBS. Demand is
highest in the USA and Europe, while the major
sources of FBS are far away from these areas — in
Brazil, Argentina, South Africa, Australia, New
Zealand, and Central America. It is in these coun-
tries that huge meat cattle herds — bulls and cows
— roam freely together, and as a result, many cows
are pregnant at the time of slaughter.
13,14
The same
holds for the geographical distances between raw
serum producers and FBS processors. The latter are
also mainly located in the USA and in Europe. It is
estimated that approximately 500,000 litres of FBS
are sold per year, which means that more than
1,000,000 unborn bovine fetuses have to be
subjected to the harvesting procedure — a fact that
raises major animal welfare concerns,
13–16
and
indeed the numbers are still increasing. As a conse-
quence, a number of strategies were developed in
terms of the Three Rs,
17
to reduce or replace the
requirement for FBS in cell culture media.
15
As well as concerns about the number of animals
required to supply the FBS market, there are addi-
tional concerns that this market is only loosely
regulated
18–21
— and this creates opportunities for
abuse.
18,19,22
This abuse has been evident in the
past, and is most likely still happening now. For
example, in 1994 it was reported
20
that approxi-
mately 30,000 litres of “New Zealand” FBS was
sold worldwide. However, only 15,000 litres of
high-quality FBS were annually collected in New
Zealand. Even now, exact figures for the global
FBS production rate are still unavailable, which
raises suspicions as to whether FBS in general
might be blended with other sera to meet
increasing demands. No attempts have ever been
ATLA 42, 207–209, 2014 207
Comment
A Severe Case of Fraudulent Blending of Fetal Bovine
Serum Strengthens the Case for Serum-free Cell and
Tissue Culture Applications
undertaken to trace the collected sera, in order to
gain clear evidence about their geographical
origin.
Obviously, in the last 20 years nothing has
changed.
18–20
As pointed out above, many FBS
batches were blended with bovine serum albumin,
water and growth promoting additives.
1
The US
Food and Drug Administration (FDA) reports that
143 batches of FBS, amounting to a total of approx-
imately 280,000 litres, are affected.
23
This latest
incident might be just the tip of the iceberg. Most
importantly, the actual case might also have a
substantial impact on many thousands of cell and
tissue culture experiments, and, in particular,
where GLP and GMP conditions are required, this
can hardly be ignored!
This recent fraudulent action should be taken as
an opportunity to question the use of FBS as cell
culture media supplement. We therefore appeal to
cell and tissue culturists to reduce or completely
avoid FBS in their cultures, and to turn to other
options, e.g. serum-free cell and tissue culture,
15,16,24
or the replacement of FBS by the use of serum
substitutes, such as human platelet lysates.
25–27
In
particular, cultures that are newly initiated should
be grown from the very beginning under serum-free
conditions. In 2003 and 2009, European cell culture
experts gathered at two workshops to discuss
options for, and the methodologies of, serum-free cell
culture. Two comprehensive workshop reports were
published,
15,16
in which clear recommendations for
the replacement of FBS, and for the design of serum-
free media, respectively, are provided. Following
these report recommendations will result in:
scientifically better (and more-reproducible) data;
safer products;
ethical research without harming animals;
better availability of cell and tissue culture
media;
the transparent and traceable composition of
culture media; and
a significant contribution to Good Cell Culture
Practice
28
(GCCP).
Professor Gerhard Gstraunthaler
Division of Physiology
Innsbruck Medical University
Innsbruck
Austria
E-mail: gerhard.gstraunthaler@i-med.ac.at
Professor Toni Lindl
Institut für Angewandte Zellkultur
Munich
Germany
Dr Jan van der Valk
3Rs-Centre Utrecht Life Sciences
Utrecht University
Utrecht
The Netherlands
References
1
Gstraunthaler, G., Lindl, T. & van der Valk, J. (2013).
A plea to reduce or replace fetal bovine serum in cell
culture media. Cytotechnology 65, 791–793.
2
Gstraunthaler, G. & Lindl, T. (2013). Zell- und
Gewebekultur. 7th edition, 340pp. Heidelberg, Germ -
any: Springer Spektrum Verlag.
3
Anderson, N.L. & Anderson, N.G. (2002). The human
plasma proteome. History, character, and diagnostic
prospects. Molecular & Cellular Proteomics 1,
845–867.
4
Anderson, N.L., Polanski, M., Pieper, R., Gatlin, T.,
Tirumalai, R.S., Conrads, T.P., Veenstra, T.D.,
Adkins, J.N., Pounds, J.G., Fagan, R. & Lobley, A.
(2004). The human plasma proteome: A non-redun-
dant list developed by combination of four separate
sources. Molecular & Cellular Proteomics 3, 311–326.
5
Psychogios, N., Hau, D.D., Peng, J., Guo, A.C.,
Mandal, R., Bouatra, S., Sinelnikov, I., Krish na -
murthy, R., Eisner, R., Gautam, B., Young, N., Xia, J.,
Knox, C., Dong, E., Huang, P., Hollander, Z.,
Pedersen, T.L., Smith, S.R., Bamforth, F., Greiner, R.,
McManus, B., Newman, J.W., Goodfriend, T. &
Wishart, D.S. (2011). The human serum metabolome.
PloS One 6, e16957.
6
Shailer, C. & Corrin, K. (1999). Serum supply: Policies
and controls operating in New Zealand. Developments
in Biological Standardization 99, 71–77.
7
Asher, D.M. (1999). Bovine sera used in the manufac-
ture of biologicals: Current concerns and policies of
the U.S. Food and Drug Administration regarding the
transmissible spongiform encephalopathies. Develop -
ments in Biological Standardization 99, 41–44.
8
Dormont, D. (1999). Transmissible spongiform
enceph alopathy agents and animal sera. Develop -
ments in Biological Standardization 99, 25–34.
9
Even, M.S., Sandusky, C.B. & Barnard, N.D. (2006).
Serum-free hybridoma culture: Ethical, scientific and
safety considerations. Trends in Biotechnology 24,
105–108.
10
Wessmann, S.J. & Levings, R.L. (1999). Benefits and
risks due to animal serum used in cell culture produc-
tion. Developments in Biological Standardization 99,
3–8.
11
Brindley, D.A., Davie, N.L., Culme-Seymor, E.J.,
Mason, C., Smith, D.W. & Rowley, J.A. (2012). Peak
serum: Implications of serum supply for cell therapy
manufacturing. Regenerative Medicine 7, 7–13.
12
Fujimoto, B. (2002). Fetal bovine serum — Supply vs
demand? Art to Science 21, 1–4.
13
Jochems, C.E., van der Valk, J.B.F., Stafleu, F.R. &
Baumans, V. (2002). The use of fetal bovine serum:
Ethical or scientific problem? ATLA 30, 219–227.
14
Brunner, D., Frank, J., Appl, H., Schöffl, H., Pfaller,
W. & Gstraunthaler, G. (2010). Serum-free cell
culture: The serum-free media interactive online data-
base. ALTEX 27, 53–62.
15
van der Valk, J., Brunner, D., De Smet, K., Fex
Svenningsen, Å., Honegger, P., Knudsen, L.E., Lindl,
T., Noraberg, J., Price, A., Scarino, M.L. & Gstraun -
208 Comment
thaler, G. (2010). Optimization of chemically defined
cell culture media — Replacing fetal bovine serum in
mammalian in vitro methods. Toxicology in Vitro 24,
1053–1063.
16
van der Valk, J., Mellor, D., Brands, R., Fischer, R.,
Gruber, F., Gstraunthaler, G., Hellebrekers, L.,
Hyllner, J., Jonker, F.H., Prieto, P., Thalen, M. &
Baumans, V. (2004). The humane collection of fetal
bovine serum and possibilities for serum-free cell and
tissue culture. Toxicology in Vitro 18, 1–12.
17
Balls, M., Goldberg, A.M., Fentem, J.H., Broadhead,
C.L., Burch, R.L., Festing, M.F.W., Frazier, J.M.,
Hendriksen, C.F.M., Jennings, M., van der Kamp,
M.D.O., Morton, D.B., Rowan, A.N., Russell, C.,
Russell, W.M.S., Spielmann, H., Stephens, M.L.,
Stokes, W.S., Straughan, D.W., Yager, J.D., Zurlo, J.
& van Zutphen, B.F.M. (1995). The Three Rs: The way
forward. The report and recommendations of ECVAM
Workshop 11. ATLA 23, 838–866.
18
Hodgson, J. (1991). Checking the sources: The serum
supply secret. Nature Biotechnology 9, 1320–1324.
19
Hodgson, J. (1993). Fetal bovine serum revisited.
Nature Biotechnology 11, 49–53.
20
Hodgson, J. (1995). To treat or not to treat: That is the
question for serum. Nature Biotechnology 13, 333–
343.
21
Nielsen, O. (1995). Changing serum’s mind-set.
Nature Biotechnology 13, 626.
22
Bohn, B. (1995). Fatal bovine serum. Nature Bio -
technology 13, 926–927.
23
FDA (2013). Class 2 Device Recall GE Healthcare/
PAA Healthcare. Silver Spring, MD, USA: US Food
and Drug Administration. Available at: http://www.
accessdata.fda.gov/scripts/cdrh/cfdocs/cfRes/res.cfm?
ID=117863 (Accessed 02.04.14).
24
Gstraunthaler, G. (2003). Alternatives to the use of
fetal bovine serum: Serum-free cell culture. ALTEX
20, 275–281.
25
Bieback, K. (2013). Platelet lysate as replacement for
fetal bovine serum in mesenchymal stromal cell
cultures. Transfusion Medicine & Hemotherapy 40,
326–335.
26
Rauch, C., Feifel, E., Amann, E-M., Spötl, H.P., Schen -
nach, H., Pfaller, W. & Gstraunthaler, G. (2011).
Alternatives to the use of fetal bovine serum: Human
platelet lysates as a serum substitute in cell culture
media. ALTEX 28, 305–316.
27
Rauch, C., Wechselberger, J., Feifel, E. & Gstraun -
thaler, G. (2014). Human platelet lysates successfully
replace fetal bovine serum in adipose-derived adult
stem cell culture. Journal of Advanced Biotechnology &
Bioengineering 2, 1–11.
28
Coecke, S., Balls, M., Bowe, G., Davis, J., Gstraun -
thaler, G., Hartung, T., Hay, R., Merten, O.W., Price,
A., Schechtman, L., Stacey, G. & Stokes, W. (2005).
Guidance on Good Cell Culture Practice. A report of
the second ECVAM task force on Good Cell Culture
Practice. ATLA 33, 261–287.
Comment 209
... FBS is a by-product of the meat industry, which can be affected by various factors, for example, geographical factors and conditions, or the amount of meat consumption in the world and the slaughters performed can change the amount of access. One of the problems that arise due to more use of FBS is the global share and income, Followed by the creation of conditions that allow for increased profits, as well as an environment conducive to fraud and the use of low-quality materials (Gstraunthaler et al. 2014;Hodgson 1995;Brindley et al. 2012). ...
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... This is not to mention the batch-to-batch variation that is frequently observed in FBS, potentially adding a factor that causes inconsistency in the cellular therapy product. The processing procedure in FBS production contributes as the key factor in controlling batch variation, while the poor regulation of FBS production may lead to cases of non-conformity [22], greatly affecting the results in cell cultures. Studies observed that the content of endotoxin and growth hormone varied widely in batches of FBS, resulting in the inconsistency in cell expansion [23,24]. ...
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... BS supply is also highly dependent on the beef consumption industry, which is itself dependent on a multitude of environmental and regulatory factors [15,16]. Ultimately, this has led to concerns about fraudulent sera blending in response to the increasing BS demand in what appears to be a loosely regulated market [17,18]. ...
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Pflanzliche Zell- und Gewebekulturen sind zu einem wichtigen Hilfsmittel der Grundlagenforschung und der praktischen Pflanzenzüchtung geworden, da ihre Relevanz für die Biotechnologie und die Agrarwirtschaft erkannt worden ist (Abb. 23-1a, Abb. 23-1b).
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Mesenchymal stromal cells (MSC) emerged as highly attractive in cell-based regenerative medicine. Initially thought to provide cells capable of differentiation towards mesenchymal cell types (osteoblasts, chondrocytes, adipocytes etc.), by and by potent immunoregulatory and pro-regenerative activities have been discovered, broadening the field of potential applications from bone and cartilage regeneration to wound healing and treatment of autoimmune diseases. Due to the limited frequency in most tissue sources, ex vivo expansion of MSC is required compliant with good manufacturing practice (GMP) guidelines to yield clinically relevant cell doses. Though, still most manufacturing protocols use fetal bovine serum (FBS) as cell culture supplement to isolate and to expand MSC. However, the high lot-to-lot variability as well as risk of contamination and immunization call for xenogenic-free culture conditions. In terms of standardization, chemically defined media appear as the ultimate achievement. Since these media need to maintain all key cellular and therapy-relevant features of MSC, the development of chemically defined media is still - albeit highly investigated - only in its beginning. The current alternatives to FBS rely on human blood-derived components: plasma, serum, umbilical cord blood serum, and platelet derivatives like platelet lysate. Focusing on quality aspects, the latter will be addressed within this review.
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