Skip to main content

Best practices for the use and evaluation of animal serum as a component of cell culture medium


Animal serum is a common additive for cell culture medium and is often required at 5 to 10% (v/v) for the attachment and growth of primary and continuous anchorage-dependent (monolayer) cultures. The use of animal serum in cell culture medium confers several advantages and also some risks. This article discusses the use of animal serum as a component of cell culture medium. The best practices associated with the sourcing, storage, thawing, testing, and mitigation of risk associated with the use of animal sera are among the topics described in this article.

This is a preview of subscription content, access via your institution.

Fig. 1.
Fig. 2.


  • Armstrong SE, Mariano JA, Lundin DJ (2010) The scope of mycoplasma contamination within the biopharmaceutical industry. Biologicals 38:211–213

    Article  CAS  PubMed  Google Scholar 

  • Barile MF, Kern J (1971) Isolation of Mycoplasma arginini from commercial bovine sera and its implication in contaminated cell cultures. Proc Soc Exp Biol Med 138:432–437

    Article  CAS  PubMed  Google Scholar 

  • Brunner D, Frank J, Appl H, Schöffl H, Pfaller W, Gstraunthaler G (2010) Serum-free cell culture: the serum-free media interactive online database. ALTEX 27:53–62

    Article  PubMed  Google Scholar 

  • Danner DJ, Smith J, Plavsic M (1999) Inactivation of viruses and mycoplasmas in fetal bovine serum using 56°C heat. Bio Pharm 12:50–52

    Google Scholar 

  • Dehghani H, Rasmussen B, Fung V, Piper R, Gosink J, Weaver B, Reddy P (2007) Case studies of mycoplasma in CHO cell cultures. In: Proceedings from the PDA Workshop on mycoplasma contamination by plant peptones. Bethesda MD; Pharmaceutical Drug Association, 53–59

  • Drexler HG, Uphoff CC (2002) Mycoplasma contamination of cell cultures: incidence, sources, effects, detection, elimination, prevention. Cytotechnology 39:75–90

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • European Medicines Agency. Guideline on the use of bovine serum in the manufacture of human biological medicinal products. EMA/CHMP/BWP/457920/2012 rev. 1, effective 2013

  • European Pharmacopoeia. Bovine serum. 01/2008:2262.

  • Hansen G, Foster L (1997) Viral safety in serum for cell culture use. Hyclone Art to Science 16(2):1–7

  • International Serum Industry Association. Frequently asked questions—bovine serum. 2011.

    Google Scholar 

  • Franke J, Abs V, Zizzadoro C, Abraham G (2014 Published online 2014 May 26) Comparative study of the effects of fetal bovine serum versus horse serum on growth and differentiation of primary equine bronchial fibroblasts. BMC Vet Res 10:119 10.1186/1746-6148-10-119

  • Gstraunthaler G (2003) Alternatives to the use of fetal bovine serum: serum-free cell culture. ALTEX 20:275–281

    PubMed  Google Scholar 

  • Nims R (2011) Adventitious viral contamination of biopharmaceuticals: who is at risk? Bioprocess J 10:4–10

    Article  Google Scholar 

  • Nims RW, Gauvin G, Plavsic M (2011) gamma Irradiation of animal sera for inactivation of viruses and mollicutes—a review. Biologicals 39:370–377

    Article  CAS  PubMed  Google Scholar 

  • Price PJ (2017) Best practices for media selection for mammalian cells. In Vitro Cell Dev Biol – Anim. doi:10.1007/s11626-017-0186-6

  • Reid YA, Baust JM, Buehring GC, Campbell LH, Elmore E, Harbell JW, Nims RW, Price P, Simione F (2017) Best practices in cell culture: an overview. In Vitro Cell Dev Biol – Anim

  • Sampath R, Blyn LB, Ecker DJ (2010) Rapid molecular assays for microbial contaminant monitoring in the bioprocess industry. PDA J Pharm Sci Technol 64:458–464

    CAS  PubMed  Google Scholar 

  • United States Pharmacopeia. <1024> Bovine Serum

  • United States Code of Federal Regulations (1995) Detection of extraneous viruses by the fluorescent antibody techniques. 9 CFR 113.47

  • United States Code of Federal Regulations (2012) Requirements for ingredients of animal origin used for production of biologics. 9 CFR 113.53

  • World Health Organization (2010) Annex 3. Recommendations for the evaluation of animal cell cultures as substrates for the manufacture of biological medicinal products and for the characterization of cell banks.

  • Yang Z, Xiong H.-R (2012) Culture conditions and types of growth media for mammalian cells. INTECH

  • Zheng X, Baker H, Hancock WS, Fawaz F, McCaman M, Pungor E Jr (2006) Proteomic analysis for the assessment of different lots of fetal bovine serum as a raw material for cell culture: part IV. Application of proteomics to the manufacture of biological drugs. Biotechnol Prog 22:1294–1300

    Article  CAS  PubMed  Google Scholar 

Download references


The authors acknowledge the input and expert review of the following members of the Best Practices in Cell Culture Workgroup: John M. Baust, Gertrude Case Buehring, Lia H. Campbell, Eugene Elmore, Paul Price, Frank Simione, and Yvonne A. Reid.

Author information

Authors and Affiliations


Corresponding author

Correspondence to Raymond W. Nims.

Additional information

Editor: Tetsuji Okamoto


Adventitious agents

Infectious agents which can contaminate cell cultures, such as viruses and bacteria.

Anchorage-dependent cells or cultures

Cells, or cultures derived from them, which will grow, survive, or maintain function only when attached to a surface such as glass or plastic. The use of this term does not imply that the cells are normal or that they are or are not neoplastically transformed.


The situation which exists when the nucleus of a cell does not contain an exact multiple of the haploid number of chromosomes; one or more chromosomes being present in greater or lesser number than the rest. The chromosomes may or may not show rearrangements.

Cell line

A cell line arises from a primary culture at the time of the first successful subculture. The term “cell line” implies that cultures from it consist of lineages of cells originally present in the primary culture. The terms “finite” or “continuous” are used as prefixes if the status of the culture is known. If not, the term line will suffice. The term “continuous line” replaces the term “established line.” In any published description of a culture, one must make every attempt to publish the characterization or history of the culture. If such has already been published, a reference to the original publication must be made. In obtaining a culture from another laboratory, the proper designation of the culture, as originally named and described, must be maintained and any deviations in cultivation from the original must be reported in any publication.

Chemically defined medium

A nutritive solution for culturing cells in which each component is of known chemical structure.


Chinese hamster ovary

Continuous cell culture

A culture which is apparently capable of an unlimited number of population doublings, often referred to as an immortal cell culture. Such cells may or may not express the characteristics of in vitro neoplastic or malignant transformation.

Dialysis/charcoal stripping

A treatment of serum intended to remove certain low molecular weight protein complexes, polar and non-polar chemicals, hormones, cytokines, glucose, amino acids, antibiotics, and other exogenous molecules (dialysis), or to remove non-polar constituents of serum such as hormones, growth factors, cytokines, and lipid-enveloped viruses (charcoal stripping).


The situation where the nucleus of the cell contains the normal (i.e., two times the haploid) number of chromosomes.


Equine (horse) bronchial fibroblast


European Medicines Agency


Fetal bovine serum


Good manufacturing practices

Heat inactivation

The process of raising the temperature of serum to 56°C for a short period of time (typically 30 min) to inactivate complement.


Immunoglobulin G

IgG stripping

A treatment of serum intended to remove undesired antibodies from the serum.


Mouse (murine) myeloma cell line


The transfer or transplantation of cells, with or without dilution, from one culture vessel to another. It is understood that any time cells are transferred from one vessel to another, a certain portion of the cells may be lost and, therefore, dilution of cells, whether deliberate or not, may occur. This term is synonymous with the term “subculture.”

Passage number

The number of times the cells m the culture have been subcultured or passaged. In descriptions of this process, the ratio or dilution of the cells should be stated so that the relative cultural age can be ascertained.

Plating efficiency

This is a term which originally encompassed the terms, “Attachment (“Seeding”) efficiency”, “Cloning efficiency,” and “Colony forming efficiency” and which is now better described by using one or more of them in its place as the term “plating” is not sufficiently descriptive of what is taking place.

Population doubling time

The interval, calculated during the logarithmic phase of growth in which, for example, 1.0 × 106 cells increase to 2.0 × 106 cells. This term is not synonymous with “cell generation time.”

Primary culture

Primary cultureA culture started from cells, tissues, or organs taken directly from organisms. A primary culture may be regarded as such until it is successfully subcultured for the first time. It then becomes a “cell line.”


Mouse (murine) myeloma cell line, multi-drug resistant.

Rights and permissions

Reprints and Permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Nims, R.W., Harbell, J.W. Best practices for the use and evaluation of animal serum as a component of cell culture medium. In Vitro Cell.Dev.Biol.-Animal 53, 682–690 (2017).

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: