Guidelines for the use of cell lines in biomedical research

Abstract

Cell-line misidentification and contamination with microorganisms, such as mycoplasma, together with instability, both genetic and phenotypic, are among the problems that continue to affect cell culture. Many of these problems are avoidable with the necessary foresight, and these Guidelines have been prepared to provide those new to the field and others engaged in teaching and instruction with the information necessary to increase their awareness of the problems and to enable them to deal with them effectively. The Guidelines cover areas such as development, acquisition, authentication, cryopreservation, transfer of cell lines between laboratories, microbial contamination, characterisation, instability and misidentification. Advice is also given on complying with current legal and ethical requirements when deriving cell lines from human and animal tissues, the selection and maintenance of equipment and how to deal with problems that may arise.

Microbial and Viral Contamination of Animal and Stem Cell Cultures: Common Contaminants, Detection and Elimination

“…The standardised procedures and training protocols should be followed strictly on a timely basis. 13,51 Improper handling usually leads to mixing and contamination, and therefore utmost care should to taken by avoiding conversations, coughing, sneezing, spilling of medium on surfaces and accidental touching of pipettes to surfaces. [52][53][54] …”

A critical analysis of cancer biobank practices in relation to biospecimen quality

“…Cancer cell lines are accessible, inexpensive and, in some cases, represent adequate disease models (Wilding and Bodmer 2014). Concerns over the impacts of cell line cross-contamination and mis-identification are now being addressed through better cell culture practices (Geraghty et al 2014) and journal-imposed requirements for researchers to authenticate cell lines using short tandem repeat (STR) profiling (Lichter et al 2010). However, other limitations, such as those imposed by routine cell culture systems, the consequences of long-term passaging, including loss of tumour heterogeneity, and genetic drift from genomic instability, result in cell lines incompletely representing the original tumour state within the host (LaBaer 2012;Wilding and Bodmer 2014).…”

“…Concomitantly with authentication becoming more frequent among the large community of researchers employing cell lines (Geraghty et al 2014;Lichter et al 2010), the potential for sample misidentification in biobanks is being recognised and acted upon, although data regarding the rates of sample misidentification are generally lacking. Sample authentication can be achieved by STR profiling of archived blood spots on Guthrie cards collected prior to sample archiving that can be assayed as required (Cardoso et al 2010 …”

Is cell culture a risky business? Risk analysis based on scientist survey data

“…Cell culture risks must be managed for in vitro work to be reproducible. Cell line repositories were established to manage these risks, ensuring that reliable cell line stocks are accessible to the research community 10 . Repositories employ a "cell banking" approach when handling a cell line, expanding the culture to give a "seed stock" or "master bank" of vials.…”

“…This approach minimizes the risks associated with prolonged passaging and allows testing of stocks for authenticity and microbial contamination 11,12 . The essentials of Good Cell Culture Practice have been clearly documented, and methods for testing contamination are becoming standardised and more accessible 10,13 . However, despite ongoing efforts, many laboratories are not aware of the common cell culture problems and do not manage the risks effectively.…”