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目的 探讨一种新的有效离散体外培养的神经干细胞球的方法。方法 采用单纯玻璃吸管机械分散,胰蛋白酶消化,不锈钢滤网研磨,单纯滤网吸管分散和滤网吸管结合短时间胰酶消化等五种方法离散神经干细胞球,比较其各自的离散效果和离散后细胞存活情况。结果 单纯玻璃吸管吹打不能完全离散神经球;胰蛋白酶消化存30min内不能完全离散神经球,延长酶消化时间虽可离散细胞,但细胞难以存活;不锈钢滤网研磨或单纯滤网吸管吹打离散对细胞损伤大,存活率低(分别为64.1％和71.9％);滤网吸管结合短时间(3~5min)胰酶消化离散细胞效果好,存活率(92.1％)显著高于上述其它方法(P<0.05)。结论 滤网吸管结合短时间胰酶消化法离散神经干细胞球,离散效果好,细胞存活率高,是一种新的有效离散体外培养的神经干细胞球的好方法。
Objective To explore a new method of effectively dispersing neural stem cells in vitro. Methods Five kinds of methods, mechanical dispersion, trypsin digestion, stainless steel mesh grinding, simple mesh pipette dispersion and mesh pipette combined with short trypsin digestion, were used to disperse neural stem cell spheres. Their discrete effects and their discrete effects were compared Cell survival. Results The glass ball could not completely separate the neurospheres. The trypsin digestion could not completely disperse the neurospheres within 30 min, but the cells were difficult to survive even though the digestion time of the enzyme was prolonged. The stainless steel mesh or pure mesh pipette was used to separate the cells (64.1% and 71.9%, respectively). The combination of filter straw and short-time trypsin digestion (3 ~ 5min) had good effect on digestion of discrete cells and the survival rate (92.1%) was significantly higher than the other methods (P < 0.05). Conclusion Mesh straw combined with short trypsin digestion method to disperse neural stem cell spheres has good discrete effect and high cell survival rate. It is a new and effective method to effectively disperse neural stem cells in vitro.