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目的 旨在以微小免疫磁珠 (平均直径≤ 5 0 nm)分离提纯 SD大鼠胚胎大脑皮层组织中的神经干细胞并进行培养 ,观察细胞的增殖分化情况并对其分化产物进行鉴定 ,评价该分离方法的可行性及其在神经干细胞研究中的应用价值。方法 取 SD大鼠的胚胎大脑皮层组织制单细胞悬液 ,用微小磁珠分选出神经巢蛋白(nestin)阳性的细胞群 ,光镜下检测其纯度及活力后进行体外培养 ,特定条件下诱导分化并以免疫细胞方法检测产物的细胞类型。结果 分离的神经干细胞纯度为 (89.2± 4.8) % ,细胞存活率 (97.0± 1 .6) % ,诱导分化产物中出现 NSE阳性、GFAP阳性、巢蛋白 (nestin)反应阳性细胞群落。结论 微小磁珠法分离提纯神经干细胞纯度高 ,获得细胞数多 ,并对细胞的生物活性无明显影响 ,提纯的细胞可用于后续实验
The aim of this study is to isolate and culture neural stem cells in the cerebral cortex of SD rat embryos by using tiny immunomagnetic beads (mean diameter ≤ 50 nm), observe the proliferation and differentiation of the cells and identify the differentiated products, and evaluate the separation Feasibility of the Method and Its Value in Neural Stem Cell Research. Methods The single cell suspension was prepared from the cerebral cortex of SD rats and the nestin-positive cells were sorted out with micro-magnetic beads. The purity and viability of the cells were detected by light microscopy and cultured in vitro. Under certain conditions Differentiation was induced and the cell type of the product was detected by immunocytochemistry. Results The purity of neural stem cells was (89.2 ± 4.8)% and the cell survival rate was (97.0 ± 1.6)%. NSE-positive, GFAP-positive and nestin-positive cell populations were found in the differentiated cells. Conclusion The separation and purification of micro-magnetic beads method of neural stem cells with high purity, the number of cells obtained, and no significant effect on the biological activity of the cells, the purified cells can be used for subsequent experiments