Isolation And Cloning Of Goat ES-like Cell

Experiments were performed to determine the effect factors of culture> clone-, isolation and passage of embryonic stem (ES) cell and embryonic germ (EG) cell in goat.Method 1: the goat embryonic stem cell-like was isolated by culture system composed of embryonic fibroblast feeder of Kun Ming strain white mouse(MEF) or embryonic fibroblast feeder of goat(GEF), medium(high gluoice DMEM, 7.5%NBS, 7.5%FCS, O.lmM 3 -mercaptoethunol, lOOIU/ml penicilin,100IU/ml streptomycin sulfate) supplied with lOng/ml SCF and 20ng/ml LIF,from 5-6th day embryos, the goat ES-like cells were cloned to 4 pas-sage, including 30 first passage ES-like cells(33.3%),8 second passage ES-like cells(8.9%),3 third passage ES-like cells(3.3%) and 2 forth passage ES-like cells(2.2%).The experiment researched the effect of embryonic age^ growth factor^ feeder cell according to isolation and clone of goat ES-like cells. The result show: the highest implantation is the embryos(77.4%); 11 ES-like cells were isolated from 5.5th day embryos and 2 ES-like cell were cloned to forth passage. The implantion percentage of embryos and the cloned ES-like cells were low. It is benefit for blastocysts attached and ICM proliferation to supplied with LIF and Insuine. The ES-like cell can be cloned with Insulin, and cloned to passage one, nor ES-like cell with LIF. There is no different between the fibroblast feeder of mouse and the fibroblast feeder of goat according to blastoysts attached and ICM proliferation. ES-like cell develop embryoid body-like structure in suspension culture in vitro. ES-like cells of goat differentiate into various kinds cell, egg: fibroblast cell, neurolike cell. The behavior of the growth is differece beweeten goat's embryos and mouse's embryos.Method2: Primordial germ cells(PGCs) were isolated from 45-70d goat embryonic genital ridges and concultured with homogenous embryonic fibroblast. medium( high glucose DMEM, 7.5%NBS, 7.5%FCS,0.1mM 3 -mercaptoethun-ol,100IU/ml penicilin,100IU/ml streptomycin sulfate) supplied with 10ng/ml SCF and 20ng/ml L1F.36 PGCs were isolated from 45d goat embryonic genital ridges and cloned to forth passage. 11 PGCs were isolated from 55d goat and cloned to seconed passage, nor PGCs from 70th day goat embryonic genital. The presence of LIF had no positive influence on the isolation of ES-like cell. Two PGCs were cloned to forth passage supplied with LIF and SCF. PGCs were cloned to third passage supplied with LIF and SCF on feeder cell. PGCs has the characterization of ES-like.