Cell Culture Quality Control

The Coriell Cell Repositories has developed and maintains preventative and surveillance procedures to guarantee that the Repository distributes highly-characterized, viable, and contaminant-free cell cultures.

All cell cultures in the NIGMS and NIA Biobanks are grown and frozen in antibiotic-free media to aid in the detection and prevention of contamination. Cultures are tested and found free of mycoplasma, bacteria, and fungi during expansion, at the time of frozen storage, and after recovery of stock for distribution from liquid nitrogen. The protocol used for detection of mycoplasma by PCR has been published [Toji, L.H., Lenchitz, T.C., Kwiatkowski, V.A., Sarama, J.A., Mulivor, R.A. Validation of routine mycoplasma testing by PCR. In Vitro Cell. Dev. Biol. Anim. 34: 356-8 (1998)] and is available.

Lymphoblast cultures are also tested for the presence of Human Immunodeficiency Virus (HIV). Recovered cultures are also examined to determine viability and growth potential and to verify species of origin. All chromosomally aberrant cell cultures, human/rodent somatic cell hybrid cultures, and normal controls are karyotyped by G-banded analysis. Hybrids are additionally analyzed by FISH with total human DNA. In some cases, alpha-satellite FISH or FISH with specific cosmid probes or chromosome-specific paint probes have also been used to characterize hybrids.

Family relationships have been verified by Southern blot hybridization with minisatellite (VNTR) probes or by PCR using a panel of microsatellite markers. Gender has been verified by PCR with a Y chromosome-specific primer pair. Replicate cultures or matched cultures of differing cell types from the same individual have been analyzed by PCR using microsatellite and Y chromosome-specific primer pairs to assure cell culture identity.

In addition, to assure the identity of cultures comprising the Repository's inherited disorders and normal variants collection, the Repository has performed appropriate assays to verify biochemical or molecular defects or utilized expert curators for this purpose. Feedback from recipients including utilization surveys and publications citing the use of Repository cultures have also been reviewed for this purpose. Relevant information from these publications has been included in the catalog.

A flow chart illustrating the steps in the quality control process for cell cultures is available. Certificates of Analysis are available for cell line samples if requested within 60 days from the date of purchase.