Effect of leukaemia inhibitory factor and different types of feeder layers on growth and pluripotent nature of embryonic stem cells from in vitro produced goat (Capra hircus) blastocysts

摘要

Feeder layer and leukaemia inhibitory factor (LIF) play an important role in maintaining the embryonic stem cells in undifferentiated state and help in their unlimited proliferation. The aim of the present investigation was to see the effect of LIF and different types of feeder layers on growth and pluripotent nature of goat embryonic stem cells. Goat blastocysts were produced in vitro by standard methods of in vitro maturation (IVM), in vitro fertilisation (IVF) and in vitro culture (IVC) techniques. Inner cell mass (ICM) cells were isolated mechanically and cultured in embryonic stem (ES) cell medium on inactivated feeder layer supplemented with or without LIF. When ICM cells were cultured on goat fibroblast feeder layer in ES cell culture medium supplemented with LIF they maintained undifferentiated state up to 15 passages. The undifferentiated ES cells showed stem cell specific morphological features, normal karyotype and expressed stem cell specific surface markers like alkaline phosphatase,TRI-1-61, TRI-1-81 and intracellular markers Oct4, Sox2 and Nanog. It could be concluded that goat embryonic stem cells were successfully cultured on feeder layers which were not of goat origin but to maintain the undifferentiated state for long time was found the best on goat fetal fibroblast with LIF.