Concerning cell cultures for biocompatibility-testing: Monitoring by DNA-fingerprinting

Cell cultures are innovative tools for e.g. biocompatibility testing of bio materials in vitro. In our studies we used fibroblast, endothelial cell and chondrocyte cultures of human origin and of the test animal species most c ommon for this purpose in vivo. Verification of the identity of these cells is obligatory for reproducibility of the tests and Valid interpretation of the results. Cultured cells have to be checked for identity, contamination s of various origins and also for genomic mutations occuring during prolong ed cultivation in vitro or due to exposition to biomaterials. Furthermore, the risk of genetic cross-contamination with other cells increases with the number of cell cultures passaged parallel in the same laboratory. Therefor e, we generated reference fingerprints of the cultures in Varying passages for comparative monitoring of cells purposed for in vitro tests. Minisatelite DNA polymorhism resulting in reproducible individual DNA finge rprints is very discriminatory and can be used for cell culture monitoring. The patterns are stable over several passages, although sudden changes did happen in two cases, i.e. loss/gain of bands or changes in band-intensity, indicating massive genomic mutations of the cultures in vitro. Influences of biomaterials on the prints could not be detected. Several tasks can be f ollowed at the same time: detection of contaminant cells, identification of these cells of primary culture origin used for in vitro testing and finall y, monitoring for eventual genomic mutations due to prolonged cultivation o r contact to biomaterials. Inconclusive results in just one of these aspect s should lead to the disqualification of the monitored cultures from usage in vitro.