Stem Cell Research (Nov 2015)

Development of buffalo (Bubalus bubalis) embryonic stem cell lines from somatic cell nuclear transferred blastocysts

  • Syed Mohmad Shah,
  • Neha Saini,
  • Syma Ashraf,
  • Manoj K Singh,
  • Radheysham Manik,
  • Suresh K Singla,
  • Prabhat Palta,
  • Manmohan Singh Chauhan

DOI
https://doi.org/10.1016/j.scr.2015.10.010
Journal volume & issue
Vol. 15, no. 3
pp. 633 – 639

Abstract

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We developed buffalo embryonic stem cell lines from somatic cell nuclear transfer derived blastocysts, produced by hand-guided cloning technique. The inner cell mass of the blastocyst was cut mechanically using a Microblade and cultured onto feeder cells in buffalo embryonic stem (ES) cell culture medium at 38 °C in a 5% CO2 incubator. The stem cell colonies were characterized for alkaline phosphatase activity, karyotype, pluripotency and self-renewal markers like OCT4, NANOG, SOX2, c-Myc, FOXD3, SSEA-1, SSEA-4, TRA-1-60, TRA-1-81 and CD90. The cell lines also possessed the capability to differentiate across all the three germ layers under spontaneous differentiation conditions.