新生大鼠视网膜干细胞分离培养及其生物学特性
The biological characteristics of isolation and culture of retinal stem cell in neonatal rat
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摘要: 目的:探讨新生大鼠视网膜干细胞的分离培养及其生物学特性。方法:采用机械酶-分离法体外扩增培养SD大鼠视网膜干细胞,观测细胞增殖能力,绘制CCK-8检测生长曲线;诱导细胞分化,免疫组织化学检测细胞巢蛋白、溴脱氧尿核苷、微管缔合蛋白-2、神经胶质原纤维酸性蛋白、胸腺糖蛋白1.1的表达。结果:原代视网膜干细胞形成大量的细胞球悬浮生长,传代后的细胞也可以形成细胞球,呈线性生长,巢蛋白和溴脱氧尿核苷表达阳性;诱导分化后的细胞部分原纤维酸性蛋白、微管缔合蛋白-2、胸腺糖蛋白1.1表达阳性。结论:新生SD大鼠视网膜干细胞可在体外分离培养和扩增,表达干细胞特性,并可向视网膜节细胞分化。Abstract: Objective:To observe the Results of isolation and culture of retinal stem cell in the neonatal rat and investigate their biological characteristics. Methods:The mechanical enzyme separation in vitro amplification of rat retinal stem cells was done,the capacity of cell proliferation and growth curve of CCK-8 detection were observed;cell differentiation was induced,then the expressions of nestin,bromodeoxyuridine,microtubule-associated protein-2,glial fibrillary acidic protein and thymus glycoprotein-1. 1 in cells were detected by immunocytochemical method. Results:The primary retinal stem cells formed a large number of cells in the ball suspended growth,the passaged cells could also form a ball of cells,and behaved as linear growth,the nestin and bromodeoxyuridine expressions were positive;glial fibrillary acidic protein,microtubule-associated protein-2,thymus glycoprotein-1. 1 were positively expressed in part of cells after induction of differentiation. Conclusions:The neonatal SD rat retinal stem cells can be isolated,cultured,and amplified in vitro,expressed the characteristics of stem cells,it can differentiate into retinal ganglion cells.
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Key words:
- stem cells /
- retina /
- cell culture techniques /
- cell differentiation
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