人胚脑神经干细胞的分离、培养和鉴定

目的：探讨人胚脑神经干细胞的体外生长特性、分化情况和培养条件，为相关实验研究提供条件。方法：利用神经干细胞的条件培养基，对从9周和12周胚龄的自然流产胎儿分离的前脑、后脑进行神经干细胞培养，并比较其生长特性。以免疫荧光细胞化学技术检测神经上皮干细胞蛋白(Nestin)和分化后神经细胞抗原的表达。结果：两胚龄前、后脑组织中均分离出神经干细胞，他们均表达Nestin抗原阳性。血清诱导分化后，表达神经元、星型胶质细胞和少突胶质细胞的特异性抗原。9周胚龄的脑组织原代培养形成细胞克隆的比例高于12周胚龄的脑组织。在条件培养基存在的情况下，后脑组织分离的神经干细胞比前脑组织来源的更容易分化。结论：从人胚脑组织中的前、后脑中分离出神经干细胞，较小胚龄的脑组织原代培养时细胞克隆的形成比例较高，后脑组织分离的神经干细胞比前脑组织来源的更容易分化。

Isolation, Cultivation and Identification of Neural Stem Cells From Human Embryonic Cerebral Tissue

Objective: To study the growth characterization, differentiation and the cultivation of neural stem cells from human embryonic cerebral tissue in vitro for furture related reseach. Methods: Forebrain and hindbrain of human fetuses about 9 weeks and 12 weeks were isolated from spontaneous abortion. Neural stem cells were cultured in a special culture medium and were compared with growth characteriation. Immunofluorescence cytochemistry was used to detect Nestin antigen of neural stem cells and specific antigen of mature neural cells after the neural stem cells were differientiated. Results: The neural stem cells with positive expression of Nestin were isolated from forebrain and hindbrain of human fetuses about 9 weeks and 12 weeks. The cells differentiated from these stem cells expressed specific antigen of neuron astrocytes and oligodendrocytes. The ratio of cell clone formation from human embryonic cerebral tissue about 9 weeks is higher than that of 12 weeks in primary culture. Under the existence of the special medium, the differentiation of neural stem cells from hindbrain is easier than that from forebrain. Conclusion: The neural stem cells can be isolated from human embryonic cerebral tissue . The ratio of cell clone formation from smaller gestational age is higher than that from elder gestational age . The differentiation of neural stem cells from hindbrain is easier than that from forebrain.