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Complete the form below and we will email you a PDF version of “Ensuring Reproducibility: Critical Cell Culture Quality Controls”
Cell culture is an essential in vitro experimental tool. An attempt to recapitulate the body in a dish, in two and three dimensions, it has provided the basis for decades of research and probably thousands of PhDs. When it goes wrong, however, whether through accident, infection, misidentification, cross-contamination or uncontrolled differentiation (for stem cells), it can be very stressful, especially in the case of longer-term experiments or when using hard-to-replace cell lines. Another important consideration is reproducibility, which is an acknowledged life sciences industry issue. A 2015 PLOS Biology study, for example, reported in an analysis of previous studies that the prevalence of irreproducible research was over 50% – equivalent to USD $28 billion per year on irreproducible preclinical research.1 Inconsistencies in cell culture approaches are a potential issue in this regard, as if cells are not maintained or used in a consistent way, or are contaminated with an infection (like mycoplasma), this can negatively impact results and make it more difficult to reproduce and/or accurately interpret data.

“Quality control (QC) is a key part of assuring the quality of outputs from any cell culture process, and is an essential part of assuring reproducibility of scientific quality in research as well as assurance of the quality and safety of cell culture-derived products,” comments Glyn N Stacey, International Stem Cell Banking Initiative, Cambridge, UK, and the Institute for Stem Cells and Regeneration and National Stem Cell Resource Centre, Chinese Academy of Sciences, Beijing, China. “These topics are currently very much in the minds of journal editors, research funders and regulators and are thus of crucial significance to researchers.”

This article will look at these different aspects of cell culture quality control and the types of protocols that can be implemented to help ensure reliable and reproducible results.

While methods for generating organoids are still evolving, presently they are providing exciting and more accurate systems that are advancing our understanding of basic organ biology and tissue regeneration. Download this handbook to learn more about optimizing organoid culture conditions, tissue-specific organoids and troubleshooting recommendations for culture.
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Short-tandem-repeat (STR) profiling is one key method being used to authenticate cell lines. The American National Standards Institute has provided protocols that can help prove authenticity for human cell lines, and the International Cell Line Authentication Committee (ICLAC) has also provided guidance on this topic. Peat suggests using STR profiling for human cell lines can be compared to those featured in Expasy’s cell line knowledge resource, Cellosaurus. Where this type of test is not available, for example when working with cell lines from other species, Stacey suggests using the COX-1 gene sequence to confirm identity at least to the level of species of origin.6 

Current glycan detection methods are plagued with high propensity for non-specific binding making them less reliable as tools. Download this app note to discover a kit that correlates well with HPLC and MS techniques and facilitates the selection of optimal clones during cell line development.
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1. Freedman LP, et al. The economics of reproducibility in preclinical research. PLOS Biol. 2015; doi: 10.1371/journal.pbio.1002165.
2. Kalia P. De-risking cell culture: Getting canny with contamination. Technology Networks. https://www.technologynetworks.com/cell-science/how-to-guides/de-risking-cell-culture-getting-canny-with-contamination-351546. Published August 03, 2021. Accessed September 27, 2021.
3. Horbach SPJM, Halffman W. The ghosts of HeLa: How cell line misidentification contaminates the scientific literature. PLOS One. 2017;12,1–16. doi: 10.1371/journal.pone.0186281
4. Masters J. End the scandal of false cell lines. Nature. 2012;492,186. doi: 10.1038/492186a.
5. American Type Culture Collection Standards Development Organization Workgroup ASN-0002. Cell line misidentification: the beginning of the end. Nat Rev Cancer. 2010;10,441–448. doi: 10.1038/nrc2852.
6. Orla O, et al. Development and implementation of large-scale quality control for the European bank for induced Pluripotent Stem Cells. Stem Cell Res. 2020;(45). doi: 10.1016/j.scr.2020.101773.
7. Stacey GN, Hawkins JR. Cell lines: Applications and biosafety. Wooley DP, Byers KB, eds. In: Biological Safety, Principles and PracticesASM Press;2016:299-326.  doi: 10.1128/9781555819637.ch14. Accessed September 27, 2021.
8. Stacey GN. Cell culture contamination. Cree IA. (Ed) In: Cancer Cell Culture. Methods and Protocols, Vol 731. Humana Press; 2011:79-91. doi: 10.1007/978-1-61779-080-5_7.  Accessed September 27, 2021.

9. Pamies D, Leist M, Coecke S, et al. Good cell and tissue culture practice 2.0 (GCCP 2.0) – Draft for stakeholder discussion and call for action. ALTEX. 2020;37(3):490-492. doi: 10.14573/altex.2007091
10. Geraghty R, Capes-Davis A, Davis J, et al. Guidelines for the use of cell lines in biomedical research. Br J Cancer 2014;111,1021–1046. doi: 10.1038/bjc.2014.166.

11. Human Tissue Authority (HTA). Codes of Practice.  https://www.hta.gov.uk/guidance-professionals/codes-practice. Updated July 20, 2021. Accessed September 27, 2021.