Central Nervous System Pluripotent Neural Stem Cells Cultured In Vitro Model And Its Preliminary Study Of The Cytological Characteristics And In Vivo Transplantation Experiments

We have successfully isolated multipotential neural stem cells by single-cell culture method from mouse fetal striatum. Clonal analysis and immunocytochemistry method proved that the cells had the property for self-renewal and can differentiate into neurons, astrocytes and oligodendrocytes by cell-type specific antibodies and the long-term cultured model of the stem cell have been established by passaged the cells for a long-time. The karyotype analysis of neurosphere show that the cell can keep diploid even passaged for 3 months, at the interphase, the chromosomes positioned like star. The cell isolated from the cortex formed neuropheres fast but couldn't adhere to the bottom easily, in contrast, the cells from hindbrain and spinal cord formed neurospheres slowly but could adhere to the culture bottom quickly and had many cells migrated from it. When embedded in collagen gel prepared from rat tail collagen, cells can migrate from the neurospheres and invade into the collagen gel, it was found that the bFGF growth factor could promote cell's migration from the neurospheres. Rat-tail collagen is not ready for the neurosphere to adhere to and can inhibit the cell's migration when it was used as a tow-dimension substrate. Electron-microscope shown there were junctions between the stem cells, and in the neurospheres, some cells had a apopotosis-like appearance and there were small vesicles in some cells. Western-blot analysis had shown that in the stem cell culture medium which had high concentration FGF, the JNK and MAPK protein kinases had a basic phosphorylated level, after stimulated by ceramide, phosphorylated level of the JNK and MAPK did not show significant difference compared to...