Generation of handmade cloned embryos from adipose tissue derived mesenchymal stem cells in goat

Abstract

The present study aims to isolate and culture adult mesenchymal stem cells (MSCs) from adult adipose tissues of goat followed by their thorough characterization and in vitro differentiation into osteocytes, chondrocytes and adipocytes. These cells were further used as donor nuclei for production of handmade cloned embryos. The goat adipose tissue-derived mesenchymal stem cells (ADSCs) were positive for the expression of CD29, CD34, CD44, CD90 and CD166 (MSCs specific positive markers) and negative for CD45 and CD71 (MSCs specific negative markers). The expression of specific markers for osteocytes (Osteopontin, Collagen I), chondrocytes (Aggrecan and Collagen II) and adipocytes (LPL, PPARα and PPARγ) was also detected from these cells. The present study reported no significant difference in cleavage, morulae and blastocyst production, when ADSCs (69.48±0.86, 30.69±1.09 and 10.96±0.82), fetal (73.3±1.26, 34.12±0.79 and 13.86±0.69) and adult (71.81±0.64, 34.07±1.00 and 11.71±0.63) fibroblasts were used as somatic donor cells. On staining with Hoechst 33342 dye, a well-developed blastocyst revealed 90–100 numbers of blastomeres under an epifluorescence microscope. In the future, the ADSCs may be used as a convenient source for autologous therapy and production of transgenic animals.