A comparative study on efficiency of adult fibroblasts and amniotic fluid-derived stem cells as donor cells for production of hand-made cloned buffalo ( XXXX ) embryos.

@entity71469 @entity69115 @entity61681 @entity6703 @entity11520
UNASSIGNED: The efficiency of two cell types, namely adult fibroblasts, and @entity6703 ( @entity6703 ) cells as nuclear donor cells for somatic cell nuclear transfer by hand-made cloning in buffalo ( @entity11520 ) was compared. The in vitro expanded buffalo adult fibroblast cells showed a typical "S" shape growth curve with a doubling time of 40.8  h and stained positive for @entity71469 . The in vitro cultured undifferentiated @entity6703 cells showed a doubling time of 33.2  h and stained positive for alkaline phosphatase, these cells were also found positive for undifferentiated embryonic stem cell markers like @entity61681 , @entity69115 and SOX-2, which accentuate their pluripotent property. Further, when @entity6703 cells were exposed to corresponding induction conditions, these cells differentiated into osteogenic, adipogenic and chondrogenic lineages which was confirmed through alizaran, oil red O and alcian blue staining, respectively. Cultured adult fibroblasts and @entity6703 cells of passages 10-15 and 8-12, respectively, were used as nuclear donors. A total of 94 embryos were reconstructed using adult fibroblast as donor cells with cleavage and blastocyst production rate of 62.8      1.8 and 19.1      1.5, respectively. An overall cleavage and blastocyst formation rate of 71.1      1.2 and 29.9      2.2 was obtained when 97 embryos were reconstructed using @entity6703 cells as donor cells. There were no significant differences (P  >  0.05) in reconstructed efficiency between the cloned embryos derived from two donor cells, whereas the results showed that there were significant differences (P  <  0.05) in cleavage and blastocyst rates between the cloned embryos derived from two donor cell groups. Average total cell numbers for blastocyst generated using @entity6703 cells (172.4      5.8) was significantly (P  <  0.05) higher than from adult fibroblasts (148.2      6.1). This study suggests that the in vitro developmental potential of the cloned embryos derived from @entity6703 cells were higher than that of the cloned embryos derived from adult fibroblasts in buffalo hand-made cloning.
