A molecular approach to the identification and individualization of human and animal cells in culture: isozyme and allozyme genetic signatures.

The electrophoretic resolution of a group of genetically monomorphic gene-enzyme systems that are developmentally and biologically ubiquitous has been used to provide a species-specific and type-specific biochemical characterization of various cultured cells. The relative mobilities of gene-enzyme systems representing nine distinct gene products from cell cultures of 25 species from Drosophila to man are presented. These isoenzymes effectively discriminate interspecies cell-to-cell contamination and almost invariably serve to identify the contaminating species. The resolution of eight polymorphic gene-enzyme systems in human cell cultures provides a virtually unique allozyme genetic signature as a monitor of intraspecies cellular contamination. The genetic signatures of 47 commonly used human cells are presented. Included in the test were seven putative HeLa (human cervical carcinoma) contaminants each of which expressed a signature identical with that of HeLa. The probability that an unrelated human cell line will have a signature identical to a typed cell is computed for each line from the genotypic frequencies at each locus in a population of cultured human cells. The gene frequencies of this cell population are comparable to the same frequencies in natural human populations. The most common human signature has a frequency (and therefore a probability) of 0.02. The majority of the 17,010 possible signatures are far less probable. A calculation of the theoretical incidence of chance matching of signatures within test groups of two or more individuals is presented. The probability of a chance match between any two randomly selected individuals is 0.004 and among five randomly selected individuals is 0.034. The allozyme genetic signature represents a definitive monitor of cell identity and is presented as a standard of cell and tissue identification for a variety of biological studies.

Enzyme polymorphisms as genetic signatures in human cell cultures.

The electrophoretic resolution of seven relatively polymorphic human gene-enzyme systems expressed in tissue culture cells can be used as a sensitive genetic monitor for intraspecific cell contamination. An identical genotype at each of the same allozyme loci provides a 95% (or greater) confindence estimate of the identity of two cultured lines, on the basis of the allelic frequencies of the seven enzyme loci in natural populations and in populations of independently derived cultured cells. Of 27 commonly used human cell lines examined, only one of 351 pairwise comparisons proved genetically indistinguishable.