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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
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Comment 209
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