VALIDATION OF DNA FINGERPRINTING IN ANIMAL CELL TECHNOLOGY: THE DIFFERENTIATION AND IDENTIFICATION OF MURINE HYBRIDOMA CLONES FOR A SINGLE FUSION AND THE STABILITY OF TRANSFORMED CELL LINES

DNA fingerprinting is under assessment for use in the quality assurance of cell stocks: providing information on the consistency and stability of cell stocks as well as screening for cross-contamination. At the European Collection of Animal Cell Cultures (ECACC) Alec Jeffreys probes 33.6 and 33.15 have been used for analysis of cell lines from a wide range of species. We have previously reported our initial assessment of DNA fingerprinting for use in the quality control of cell banks [1, 2]. This report describes further validation of fingerprinting in animal cell technology. DNA fingerprints have demonstrated consistency between human and peripheral blood mononuclear cells and Epstein-Barr virus transformed B lymphoblastoid lines from the same individual. Analysis of long-term cultures of these cell lines (ie up to 6 months) also revealed the stability of their fingerprints. Analysis of murine hybridomas has shown certain limitations of the 33.6 probe in that clones from the same fusion expressing different Ig subclasses, could not be differentiated. Significantly probe 33.15 enabled differentiated of clones producing antibodies of identical heavy chain subclass although it was not possible to differentiate all clones. The work discussed in this report represents the ongoing process of validation of DNA fingerprinting: supporting its use in the authentication of EBV transformed cell lines and the analysis of murine hybridomas.