A multiplex PCR assay utilizing eight human short tandem repeat (STR) markers was developed for the authentication of vervet monkey cell lines. Labeled forward primers were used to amplify the targeted STR marker region and amplified products were separated using a 16-capillary electrophoresis instrument. Allele distributions and heterozygosity values were obtained from analyzing sixty-two unrelated vervet monkey samples, all resulting in unique genetic profiles. Correlations between allele fragment length and repeat number were confirmed with sequencing. STR markers have shown genetic stability in Vero cells with passage numbers ranging from seven to sixty-nine. Cell line contaminants could also be identified in a genetic profile of Vero cells when mixture samples were analyzed, suggesting the assay could be used to screen vervet cell lines for vervet and human cell line contamination. A vervet multiplex assay based on eight STR markers was successful in providing profiles unique to sixty unrelated vervet monkeys and the parent vervet cell lines, Vero and CV-1.
Citation: Bmc Biotechnology
Pub Type: Journals
"cell line authentication", "cell identification", "vervet", "African green monkey", "Vero", "short tandem repeat markers (STR)", "multiplex PCR assay", "human cell line contamination"