Genetic characterization of skin fibroblast cell lines by DNA fingerprinting
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.
Arat S, Rzucidlo SJ, Gibbons J, Miyoshi K and Stice SL (2001). Production of transgenic bovine embryos by transfer of transfected granulosa cells into enucleated oocytes. Molecular Reproduction and Development 60:20-26.
Armour JA, Tael I, Thein SL, Fey MG and Jeffreys AJ (1989). Analysis of somatic mutations in human microsatellite loci in tumour cell lines. Genomics 4:328-334.
Baker TK, Carfagna MA, Gao H, Dow ER, Li Q, Searfoss GH and Ryan TP (2001). Temporal gene expression analysis of monolayer cultured rat hepatocytes. Chemical Research in Toxicology 14:1218-1231.
Beckmann JS and Weber JL (1992). Survey of human and rat microsatellites. Genomics 12:627:631.
Boyer JC, Yamada NA, Roques CN, Hatch SB, Riess K and Farber RA (2002). Sequence dependent instability of mononucleotide microsatellites in cultured mismatch-repair-proficient and deficient mammalian cells. Human Molecular Genetics 11:707-713.
Crawford AM, Dodds KG, Ede AJ, Pierson CA, Montgomery GW, Garmonswa HG, Beattie AE, Davies K, Maddox JF and Kappes SW (1995). An autosomal linkage map of sheep genome. Genetics 140:703-724.
Enright BP, Jeong BS, Yang X and Tian XC (2003). Epigenetic characteristics of bovine donor cells for nuclear transfer : Levels of histone acetylation. Biology of Reproduction 69:1525-1530.
Farber RA, Petes TD, Dominska M, Hudgens SS and Liskay RM (1994). Instability of simple sequence repeats in a mammalian cell line. Human Molecular Genetics 3:253-256.
Geraghty RJ, Davis A Capes, Davis JM, Downward J, Freshney RI, Knezevic I, Badge R Lovell, Masters JRW, Meredith J, Stacey GN, Thraves P and Vias M (2014). Guidelines for the use of cell lines in biomedical research. British journal of cancer 111(6):1021-1046
Gupta N, Sharma R, Taneja R, Singh HN, Tantia MS, Singh G and Gupta SC (2003a). Use of somatic cells in conservation of endangered breed of Bhadawari buffaloes. Proceedings of 4th Asian Buffalo Congress held at New Delhi from 25th-28th Feb.
Gupta SC, Gupta N, Sharma R, Taneja R, Sisodia B and Bulandi SS (2003b). Manual on Somatic Cell Technology for Conservation and Regeneration of Endangered Breed. National Bureau of Animal Genetic Resources (ICAR) Publication.
Hammond K (1998). Animal genetic resources for the twenty-first century. Acta Agriculturæ Scandinavica 28:11-18.
Hirayu H and Dere WH (1987). Rapport B. Initiation of normal thyroid cells in primary culture associated with enhanced c-myc messenger ribonucleic acid levels. Endocrinology 120:924-928.
Heath DD, Iwama GK and Devlin RH (1993). PCR primed with VNTR core sequences yields species specific patterns and hypervariable probes. Nucleic Acids Research 21: 5782-5
Jeffreys AJ, Wilson V and Thein SL (1985). Individual specific fingerprints of human DNA. Nature 316:76-79.
Kubota C, Yamakuchi H, Todoroki J, Mizoshita K, Tabara N, Barber M and Yang X (2000). Six cloned calves produced from adult fibroblast cells after long-term culture. Proceedings of the National Academy of Sciences of the United States of America 97:990-995.
Liu L, Shin T, Pryor JH, Kraemer D and Westhusin M (2001). Regenerated bovine fetal fibroblasts support high blastocyst development following nuclear transfer. Cloning 3:51-58.
Nadir E, Margalit H, Gallily T and Ben-Sasson SA (1996). Microsatellite spreading in the human genome: Evolutionary mechanisms and structural implications. Proceedings of the National Academy of Sciences of the United States of America 93:6470-6475.
Ramya R, Nagarajan T, Sivakumar V, R Senthilkumar L, Bala B, Obulapathi, Thiagarajan D and Srinivasan VA (2009). Identification of cross-contaminated animal cells by PCR and isoenzyme analysis. Cytotechnology 61(3):81-92
Roh S, Shim H, Hwang WS and Yoon JT (2000). In vitro development of green fluorescent protein (GFP) transgenic bovine embryos after nuclear transfer using different cell cycles and passages of fetal fibroblasts. Reproduction, Fertility and Development 12:1-6.
Singh MK (2004). Evaluation of skin fibroblast cell line of Bhadawari buffaloes for their utility in cryopreservation and cloning. MVSc Thesis submitted to JNKVV, Jabalpur, India.
Stacey GN, Bolton BJ, Morgan D, Clark SA and Doyle A (1992). Multilocus DNA fingerprints analysis of cell banks: Stability studies and culture identification in human B lymphoblasted and mammalian cell lines. Cytotechnology 8:13-20.
Stacey GN, Hoelzl H, Stehenson JR and Doyle A (1997). Authentication of animal cell cultures by direct visualization of DNA, aldolase gene PCR and isoenzyme analysis. Biologicals 25:75-85.
Steffen P, Eggen A, Dietz AB, Womack JE, Stranzinger G and Fries R (1993). Isolation and mapping of polymorphic microsatellites in cattle. Animal Genetics 24(2):121-124.
Taneja R (2004). Cell cycle synchronization and telomere dynamics in Gaddi goat skin fibroblast in culture. PhD Thesis submitted to Kurukshetra University, Kurukshetra.
Tautz D (1989). Hypervariability of simple sequences as a general source for polymorphic DNA marker. Nucleic Acids Research 17:6463-6471.
Thacker JM, Webb MJK and Debenham PG (1988). Fingerprinting cell lines: Use of human hypervariable DNA probes to characterize mammalian cell cultures. Somatic Cell and Molecular Genetics 14:519-525.
Wilmut I, Schnieke AE, McWhir J, Kind AJ and Campbell KH (1997). Viable offspring derived from fetal and adult mammalian cells. Nature 385:810-813.