Title: Genetic characterization of skin fibroblast cell lines by DNA fingerprinting
Authors: NK Nagpal, BK Beniwal, GC Gahlot and SC Gupta
Source: Ruminant Science (2016)-5(2):149-158.
Cite this reference as: Nagpal NK, Beniwal BK, Gahlot GC and Gupta SC (2016). Genetic characterization of skin fibroblast cell lines by DNA fingerprinting. Ruminant Science 5(2):149-158.
Present study was carried out for genetic characterization of Bhadawari buffalo skin fibroblast cell lines using ten microsatellite primers. The primary cells explants from tissue of six cell lines MF-22, MF-37 and MF-82 (females) and MM-31, MM-32 and MM-79 (males) collected from 6-8 months old calves of Bhadawari buffalo breed were employed for development of cell lines. To establish the integrity of cell line which may have lost due to improper passaging, sharing of cultural media or inaccurate labeling, DNA fingerprinting was exercised. The PCR and denaturing sequence gel electrophoresis were carried out for amplification and separation of the amplified products for ten microsatellite markers. The allele size recorded for different microsatellite primers were used for the detection of cross contamination among cell lines. Out of six, three cell lines MM-31, MM-79 and MF-82 were found to be pure for their genetic integrity and were suitable for further use in somatic cell nuclear transfer. While in other three cell line MF-22, MF-37 and MM-32 intermixing was detected and should be rejected.
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