纤维连接蛋白对人神经干细胞诱导分化为少突胶质前体细胞的影响  

Effect of fibronectin on differentiation of human neural stem cells into oligodendrocyte precursor cells

作　　者：汪兆艳 王倩 刘卫鹏 杨辉 栾佐 屈素清 Wang Zhaoyan;Wang Qian;Liu Weipeng;Yang Hui;Luan Zuo;Qu Suqing(Department of Pediatrics,Sixth Medical Center of PLA General Hospital,Beijing 100048,China)

机构地区：[1]中国人民解放军总医院第六医学中心儿科,北京市100048

出　　处：《中国组织工程研究》2025年第31期6661-6666,共6页Chinese Journal of Tissue Engineering Research

摘　　要：背景:少突胶质前体细胞是治疗脑白质损伤性疾病的种子细胞,建立高效稳定扩增的体外诱导分化方法是实现临床转化研究的重要前提。目的:探讨纤维连接蛋白对人神经干细胞来源少突胶质前体细胞增殖、迁移和分化等生物学特性的影响。方法:将悬浮培养的人神经干细胞用Accutase消化成单细胞,通过流式细胞术检测特异性标志物Nestin、Sox2、Vimentin、CD133、Musashi的表达。然后将人神经干细胞单细胞重悬于少突胶质前体细胞培养基,接种于不同质量浓度纤维连接蛋白(0,1,2.5,5,10μg/mL)包被的6孔板中,培养7 d后Accutase消化进行传代,锥虫蓝染色计数细胞,选择扩增能力最强的纤维连接蛋白包被与未用纤维连接蛋白包被的少突胶质前体细胞进行下一步实验,Transwell小室检测细胞迁移能力,流式细胞术检测Olig2、Sox10、PDGFR-α的表达。将少突胶质前体细胞向少突胶质细胞诱导分化3周,免疫荧光染色检测分化细胞Galc的表达。结果与结论:①人神经干细胞呈悬浮球状生长,流式细胞术检测结果显示,人神经干细胞高表达Nestin、Sox2、Vimentin、CD133和Musashi;②由人神经干细胞诱导的少突胶质前体细胞的胞体为圆形或椭圆形,折光性强,有双极或三级突起结构;与0μg/mL纤维连接蛋白包被组比较,2.5,5,10μg/mL纤维连接蛋白包被组少突胶质前体细胞的扩增能力显著增强(P<0.05);当纤维连接蛋白质量浓度为10μg/mL时,少突胶质前体细胞扩增能力最强;③流式细胞术检测结果显示0,10μg/mL纤维连接蛋白包被组均高表达少突胶质前体细胞标志物Olig2、Sox10和PDGFR-α,两组无统计学差异(P>0.05);④Transwell小室检测结果显示,与0μg/mL纤维连接蛋白包被组比较,10μg/mL纤维连接蛋白包被组少突胶质前体细胞的迁移能力增加(P<0.01);⑤少突胶质前体细胞向少突胶质细胞分化3周,细胞呈多分支、网格状或膜片样复杂�BACKGROUND:Oligodendrocyte precursor cells are seed cells for the treatment of white matter damage diseases.Establishing an efficient and stable in vitro differentiation method is an important prerequisite for clinical translational research.OBJECTIVE:To investigate the effect of fibronectin on biological characteristics such as proliferation,migration,and differentiation of oligodendrocyte precursor cells derived from human neural stem cells.METHODS:Human neural stem cells cultured in suspension were digested into single cells using Accutase.The expression of specific markers Nestin,Sox2,Vimentin,CD133,and Musashi was detected by flow cytometry.The single cells of human neural stem cells were resuspended in oligodendrocyte precursor cell medium and seeded in six-well plates coated with different concentrations of fibronectin(0,1,2.5,5,and 10μg/mL).Accutase digestion was performed after 7 days of culture.Cells were counted by trypan staining.Fibronectin-coated group with the strongest amplification ability and the oligodendrocyte precursor cells without fibronectin-coated group were selected for further tests.The migration ability of the two groups of cells was detected by Transwell.Flow cytometry was used to detect the expression of Olig2,Sox10,and PDGFR-α.Oligodendrocyte precursor cells were induced to differentiate into oligodendrocytes for 3 weeks,and the expression of Galc in differentiated cells was detected by immunofluorescence staining.RESULTS AND CONCLUSION:(1)Human neural stem cells grew in suspension spheres.Flow cytometry showed that human neural stem cells highly expressed Nestin,Sox2,Vimentin,CD133,and Musashi.(2)The cell bodies of oligodendrocyte precursor cells induced by human neural stem cells were round or oval,with strong refractive nature and bipolar or tertiary protrusions.Compared with the 0μg/mL fibronectin coating group,there was a significant difference in the amplification ability of oligodendrocyte precursor cells in the 2.5,5,and 10μg/mL fibronectin coating groups(P<0.05).The amp

关 键 词：纤维连接蛋白 神经干细胞 少突胶质前体细胞 少突胶质细胞 细胞增殖 细胞分化 工程化干细胞 工程化细胞 

分 类 号：R459.9[医药卫生—治疗学] R318[医药卫生—临床医学] R322.8