The Study Of Purification LIF And The Suppression Mes Cell Differentiation By In Vitro

Embryonic stem (ES) cells are able to self-renewal and maintain pluripotent in special media culture condition in vitro. ES cells can also generate cells with three germ-layer derivatives while in differentiation culture conditions. The tranditional culture medium for murine embryonic stem cells uses LIF (Leukemia inhibitory factor) as a key factor to prevent the murine embryonic stem cells to differentiate. While LIF is such an expensive factor monopolized by foreign biological company and it is a serious problem for the investigators in Chinese for a long time. In order to resolve the problem we use pGEX-2T-MLIF transformed Escherichia coli to produce LIF induced by IPTG and then purify it. We then added this LIF into the stem cell culture medium and the experiments show that the LIF produced by us has the same capacity to prevent the ES cell to differentiate and maintain self-renew just like the commercialized LIF. Immuno- histochemistry experiments show that the ES cells also express pluripotent markers such as Oct4, Sox2, Nanog. We injected the ES cells to the nude and got the teratoma 4 weeks later. It proves that the LIF has biologic active and can be used in mES cell culture to maintain pluripotent.