Comparison Of The Rate Of Induced Differentiation Of Neural Stem Cells Which Are From Different Parts Of The Neonatal Rats Brain To Oligodendrocytes Cells

【Objective】To obtain and culture neural stem cells (neural stem cells, NSCs) of the three different parts of the neonate SD rat brain which is the cerebral cortex, striatum and hippocampus. To compare the multiplication and passage capability and differentiation rate of being induced into oligodendrocytes of the neuural stem cells derived from different locations of the neonate SD rat brain. To provide the theoretical bases of getting more oligodendrocytes.【Methods】1.The cells isolated from cerebral cortex, hippocampus and striatum of 24h age neonate rat brain were cultured under the effect of basic fibroblast growth factor (bFGF) and B27.The rat neural stem cell sphere differentiation was induced by 10% fetal calf serum.Technique of single-cell clone was used to confirm the character of self-renewing.The rat neural stem cells and the differentiated neural stem cells were identified respectively by immunofluorescence including Nestin, neuron specific enolase (NSE), gilial fibrillary acidic protein (GFAP) and Galc- C.2.To subculture the neural stem cells from different locations of the neonate SD rat brain.CCK8 was used to compare the proliferation capability of the NSCs to draw the cell growth curve; To compare the multiplication and passage capability the neuural stem cells derived from different locations of the neonate SD rat brain.3.To induce differentiation of NSCs by using insulin-like growth factor -1, thyroid hormone, nerve glioblastoma supernatant,used the laser confocal microscope and Western blot method to detect the cell differentiation rate after the induction, To analysis the differences of the rate of NSCs differentiation into oligodendrocytes.【Results】1.Using serum-free culture medium can obtain high-purity primary rat NSCs; the NSCs were nestin-positive and have the ability to differentiation into oligodendrocytes, neurons and astrocytes.Single-cell clone experiments can get the cells cloning ball.2. The neural stem cells derived from cerebral cortex can9-12times, 8-11 times from hippocampus while striatum can 6-9 times. CCK8 was used to draw the cell growth curve,statistical analysis : the multiplication of the hippocampus and the striatum have no significant difference (P> 0.05); the cortex and the hippocampus have significant difference (P <0.05). cortex and striatum of the proliferation activity of NSCs have significant difference (P <0.05). With the passage and the increase in the number, the differences are still maintained.3. There was no significant difference between Insulin-like growth factor -1, thyroxin of induced ability (P> 0.05); The nerve glioblastoma supernatant and insulin-like growth factor-1 have significant difference (P< 0.05) The nerve glioblastoma culture supernatants and the ability of thyroxine-induced was significantly difference (P <0.05).cortical source of neural stem cells were significantly different from the hippocampus and striatum cells to induced into oligodendrocytes (P <0.05).【Conclusion】1.Using serum-free medium suspension culture can obtain the primary NSCs which has the ability of high-purity self-renewal and multilineage differentiation.2. The neural stem cells derived from cerebral cortex have stronger passage and multiplication capacity than that from hippocampus and striatum and with the passage the differences are still maintained.3. The ability of nerve glioblastoma supernatants induced neural stem cells into oligodendrocyte is stronger than the insulin-like growth factor -1, thyroxin; the ability of cortical source of neural stem cells be induced into oligodendrocyte was stronger than hippocampus and striatum cells.