| Poluripotent stem cell derived from the embryo have attracted very close attention. When cultured in vitro, embryo-derived stem cell possess the capability of proliferation and differentiating to some special cells. Using the technique related to the embryo-derived stem cell, it is possible to develop a clone animal, which possesses the same genotype as well as the phenotype. It is also be possible to develop introduced the exogene to develop the transgenic, which could be expressed stably. The extensive application with embryo-derived stem cell can largely promote the research fields about establishing the animal modle of human disease, the medical protein production, and the clinical organ transplantation or repairing tissue.It is known to all, the extensive sever burned patient always facing to the problem of lack of auto skin. It would open a new medical area if the skin tissue cell can be developed from the stem cell through induced differentiation. At present the research of the human embryo-derived stem cell is still at the early stage. The mouse was used to got some experiences in favorite to the human embryo- derived stem cell in our study.The primordial germ cells (PGCs), which is known to be poluripotent, were isolated from the genital ridge of mouse 10 days post coitum (dpc) under the anatomic microscope. The feeder layer cells were made by the primordial mouse embryonic fibroblasts (PMEF). The cultured medium was composed of a -MEM, 15% NBS, 0.1 mM beta-mercaptocthanol, ImM sodium pyruvate, 100 lU/ml bFGF, 1M non essential amino acids, ImM L-glutamin, or added with the PMEF conditioned medium. The Percoll cell separating liquid was used when the stem cell clones were dissaggragating manually. The stem cells were identified through being observed the morphology under the invert phase contrast microscope and stained by the alkali phosphatase (AP). The stem cell were induced to differentiated with non serum culture medium, or prolonging the interval of changing the medium. The cells proliferation curve of the PMEF was checked after treated with mitomycine C of different concentration.The PGCs of 140 embryos were isolated from 80 mice. 5 stem cell lines being subcultured to the passage 3 were obtained. The active mobility of the stem cell was observed, which was positive stained with the AP. The PMEF conditioned medium delayed the survival time of the stem cell from the fourth day to the sixth day after being plated. It was showed by the concentration screening test that 10 ug/ml mitomycine C could suppress fairly the proliferation of the PMEF, which would be in a stable cell number until the fourth day after the treatment. In our study, thepoluripotent stem cells were induced to several kinds of cell, such as neurocyte, cardiac muscle cell, endothelial cell and epidermic-cell-like cell, which were identified through the morphology. Conlusins:1. Using the PMEF as feed layer, or the PMEF conditioned medium , could increase the survival time of the embryo-derived stem cell.2. 10 ug/ml mitomycine C could prepared prepaired the feeder layer cell preferably.3. Combining the percoll with dissagragating clone manually to subculture the embryo-derived stem cell is easier to do in the common laboratory than traditional method.4. The results of the study showed that the embryo-derived stem cells possess the capability to be induced to differentiated to skin tissue cells in vitro.
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