Putative amniotic fluid stem (AFS) cells express transcription factor Oct-4 in goat (Capra aegagrus hircus)

Authors

  • Sumita Verma Department of Biotechnology, Kurukshetra University, Kurukshetra-136119 Haryana India. Author
  • Kapil Dev Department of Biotechnology, Kurukshetra University, Kurukshetra-136119 Haryana India. Author
  • Shiv K. Giri Department of Biotechnology, Kurukshetra University, Kurukshetra-136119 Haryana India. Author
  • Anil Khuttan Department of Biotechnology, Kurukshetra University, Kurukshetra-136119 Haryana India. Author
  • Anita Yadav Department of Biotechnology, Kurukshetra University, Kurukshetra-136119 Haryana India. Author
  • Ashutosh Mangotra Department of Biotechnology Engineering, U.I.E.T. Kurukshetra University, Kurukshetra-136119 Haryana India. Author
  • Birbal Singh Regional Station, Indian Veterinary Research Institute, Palampur-176061 H.P. India. Author
  • Sanjeev K. Gautam Department of Biotechnology, Kurukshetra University, Kurukshetra-136119 Haryana India. Author

Keywords:

Goat, amniotic fluid, stem cells, characterization, RT-PCR

Abstract

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 156bp 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.

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Published

01-06-2012

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Articles