节点文献
酵素-Ⅱ分离人神经干细胞球
Effects of dispaseⅡ on isolating neurospheres
【摘要】 目的:寻找一种有效分离神经干细胞球并获得大量单个神经干细胞的方法。方法:取20周胎龄人胚胎脑组织分离单细胞,应用DMEM/F12培养基培养形成神经干细胞球,取神经干细胞球,分别以2.50g/L、1.25g/L胰蛋白酶及0.6u/ml、1.2u/ml、1.8u/ml、2.4u/ml酵素-Ⅱ(DispaseⅡ)消化,观察消化过程及细胞形态,对消化后的单个细胞进行培养,观察细胞生长情况,并进行免疫细胞化学染色鉴定。结果:各浓度Dispase-Ⅱ均能较完全地消化神经干细胞球,其中1.2u/ml、1.8u/mlDispase-Ⅱ消化后的细胞生长状况最佳。胰蛋白酶消化后细胞有较大的损伤,成活细胞少。消化后的细胞Nestin抗体染色结果(+),证实为神经干细胞。结论:一定浓度的DispaseⅡ消化神经干细胞球后,可获得大量成活单个神经干细胞。
【Abstract】 Aim: To investigate an effective method to isolate neurospheres. Methods: Neurospheres were isolated with different dose of trypsinase and dispaseⅡ. The process and the shape of cells were observed.Immunocytochemistry was used for identification. Results: DispaseⅡ could isolate neuropheres into single cells, especially in concentration of 1.2 u/ml and 1.8 u/ml.Cells were damaged after isolated by using trypsinase and few viable cells were obtained. Conclusion: It is an ideal method to isolate neurospheres by using a certain concentration of dispaseⅡ.
- 【文献出处】 郑州大学学报(医学版) ,Journal of Zhengzhou University(Science Medical) , 编辑部邮箱 ,2005年03期
- 【分类号】R329.2
- 【被引频次】4
- 【下载频次】113