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目的:脐血中分离CD29+细胞并诱导其转化为神经元样细胞,为神经损伤与修复提供种子细胞来源。方法:免疫磁珠法分离出脐血CD29+细胞,体外培养扩增。取2代细胞以BDNF、GDNF、PDGF为诱导因子对其进行神经诱导分化。观察细胞形态、生长情况,流式细胞仪检测细胞周期、表型及HLA相关抗原。诱导后免疫组织化学染色法检测Nestin、NSE、NF、GFAP表达。Western Blot检测β-tubulin、Tau的表达。结果:细胞贴壁生长,呈长梭形,细胞表型为CD29+、CD44+、CD166+。在诱导后,细胞逐渐增大并伸出明显突起。免疫组织化学染色结果显示,Nestin、NSE、NF、GFAP表达阳性。Western Blot显示了相应蛋白的表达。结论:脐血CD29+细胞在体外可以向神经元样细胞转化,有望成为异体修复神经损伤的种子细胞来源。
OBJECTIVE: To isolate CD29 + cells and transform them into neuron-like cells in umbilical cord blood to provide seed cell sources for nerve injury and repair. Methods: Cord blood CD29 + cells were isolated by immunomagnetic beads method and cultured in vitro. The second passage cells were induced by BDNF, GDNF and PDGF. Cell morphology and growth were observed. Cell cycle, phenotype and HLA-related antigen were detected by flow cytometry. Immunohistochemical staining was used to detect the expression of Nestin, NSE, NF and GFAP. Western Blot detection of β-tubulin, Tau expression. Results: The cells grew adherently and grew in long fusiform form. The phenotypes of the cells were CD29 +, CD44 + and CD166 +. After induction, the cells gradually increased and conspicuously protruded. Immunohistochemical staining results showed that Nestin, NSE, NF, GFAP expression was positive. Western Blot shows the expression of the corresponding protein. CONCLUSION: Cord blood CD29 + cells can be transformed into neuron-like cells in vitro and are expected to be the source of seed cells for allogeneic repair of nerve injury.