Biosafety Recommendations on the Handling of Animal Cell Cultures
The first steps in tissue culture are dating back to the beginning of the nineteenth century when biosafety measures did not yet exist. Later on, animal cell culture became essential for scientific research, diagnosis and biotechnological activities. Along with this development, biosafety concerns have emerged pointing to the risks for human health and in a lesser extent for the environment associated to the handling of animal cell cultures. The management of these risks requires a thorough risk assessment of both the cell cultures and the type of manipulation prior the start of any activity. It involves a case-by-case evaluation of both the intrinsic properties of the cell culture genetically modified or not and the probability that it may inadvertently or intentionally become infected with pathogenic micro-organisms. The latter hazard is predominant when adventitious contaminants are pathogenic or have a better capacity to persist in unfavourable conditions. Consequently, most of the containment measures primarily aim at protecting cells from adventitious contamination. Cell cultures known to harbour an infectious etiologic agent should be manipulated in compliance with containment measures recommended for the etiologic agent itself. The manipulation of cell cultures from human or primate origin necessitates the use of a type II biosafety cabinet. The scope of this chapter is to highlight aspects relevant for the risk assessment and to summarize the main biosafety recommendations and the recent technological advances allowing a mitigation of the risk for the handling of animal cell cultures.
KeywordsBiosafety Risk assessment Risk management Contained use Genetically modified organisms Pathogenic organisms
Genetically modified organism
Transmissible spongiform encephalopathies
The authors are grateful to their colleagues Dr. Didier Breyer and Dr. Aline Baldo (Scientific Institute of Public Health WIV-ISP, Brussels, Belgium) for their useful review of this manuscript.
Is the process by which the true origin and identity of cell lines are determined and should form an essential part of any cell culture operation.
In the context of this chapter, biosafety relates to the evaluation of the potential risks to human health and the environment associated with the use of genetically modified organisms (GMOs) or pathogenic organisms.
Safety cabinet with a front aperture through which the operator can carry out manipulations inside the cabinet and which is constructed so that the laboratory worker is protected, the product and cross contamination is low. The escape of airborne particulate contamination generated within the cabinet is controlled by means of an appropriate filtered internal airflow and filtration of the exhaust air (HEPA filters).
Any activity in which (micro)-organisms are genetically modified or in which such organism (pathogenic or not) are cultured, stored, transported, destroyed, disposed or used in any other way, and for which specific containment measures are used to limit their contact with the general population and the environment.
: Primary Cell Cultures, Diploid Cell Lines, Continuous Cell Lines
Are established directly from tissues of animals and are often the most appropriate in vitro tool for reproducing typical cellular responses observed in vivo. However, as typical cell characteristics are lost during passaging, these cultures must be obtained from fresh tissue that may contain or may become inadvertently contaminated with pathogens. Consequently, primary cell cultures may potentially present increased risks compared to continuous, established cell lines.
Are similar to primary cells, are considered non-tumourogenic and have a finite capacity for serial propagation. They are used for the preparation of viral vaccines and are from human or monkey origin.
Are immortalized cells that may survive almost infinite serial passages. These cells are obtained by either isolating cells from tumours (neoplastic origin), primary cells treated with mutagens, oncogenic viruses or recombinant DNA (oncogenes) or by cell fusioning of primary cells with a continuous cell line. As a consequence, the class of risk of these cell lines is often correlated to the class of risk of primary cells of whom they are derived. Due to the “immortality” of the continuous cell lines, their ability to induce tumours has also to be considered.
Gene transfer of DNA in eukaryotic cells using non-viral delivery methods.
Gene transfer of DNA in eukaryotic cells using viral vectors.
Process occurring when a virus (wild type) that typically replicates and spreads into neighbouring cells mediates a gene transfer belonging to its own genome or extra genes it may carry.
Is a protein particle derived from a replicative virus that contains genetic information in the form of RNA or DNA. A viral envelope may be present as well.
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