Author : Sumita Verma, Kapil Dev, Shiv K. Giri, Anil Khuttan, Anita Yadav, Ashutosh Mangotra, Birbal Singh and Sanjeev K. Gautam

The current study was carried out to isolate, culture, characterize and cryopreserve the putative amniotic fluid stem (AFS) cells in goat. Putative AFS cells were cultured without feeder cells, in DMEM containing 10% FBS, 1% penicillin/streptomycin and 1% L-glutamine in 5% CO2 in humidified air at 39.1±0.5˚C. After 6 days of culture different morphologies of cells were observed. Most of the cells started anchorage-dependent growth after day 6 of the culture. In order to check their pluripotency properties when the cells were subjected to RT-PCR a strong positive expression of Oct-4 was observed. Using species-specific primers, a PCR amplicon of 156 bp was observed for Oct-4. When the cells were cryopreserved in DMSO containing freezing media, >70% of the cells were viable. These results may contribute towards establishing non-embryonic pluripotent stem cells for various therapeutic and reproductive biotechnological applications in this species.

Key words: Goat, amniotic fluid, stem cells, characterization and RT-PCR.

This article was published in International Journal of  Cell Science and Biotechnology, Vol.1, 2012