Dhruba Malakar

Embryonic Stem Cell-like Cells in Goat (Capra Hircus): Derivation, Culture and Characterization Dhruba Malakar

The objective of the present experiment was production of goat embryos either by parthenogenetic activation or by in vitro fertilization and derivation of embryonic stem (ES) cell-like cells. Firstly, the effects of different activation methods, culture systems and culture media on in vitro development of zona-free and with-zona parthenogenetic embryos were investigated. Secondly, effect of cysteamine supplementation in the in vitro maturation (IVM) or culture (IVC) media, or a combination of both in terms of blastocyst production by in vitro fertilization was investigated. Finally, different methods of isolation of ES cell-like cells from in vitro produced embryos were compared and effects of Leukaemia Inhibitory Factor (LIF) and different types of feeder layers on growth of ES cell-like cells were investigated. The undifferentiated ES cell-like cells showed stem cell like morphological features, normal karyotype, expressed stem cell specific surface and intracellular markers and maintained undifferentiated state for long periods in culture. Mechanical isolation of ICMs from hatched blastocysts was found to be the best method to establish goat ES cell-like cells.