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刘 然,范东艳,金 鹏,范洪学,王 苹.细胞共培养体系在观察骨髓间充质干细胞对损伤脊髓神经元生长存活影响中的应用[J].中国康复医学杂志,2011,26(2):116~119
细胞共培养体系在观察骨髓间充质干细胞对损伤脊髓神经元生长存活影响中的应用    点此下载全文
刘 然  范东艳  金 鹏  范洪学  王 苹
吉林大学公共卫生学院,长春,130021
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摘要:
      摘要 目的:建立细胞共培养体系,观察骨髓间充质干细胞(BMSCs)对正常及损伤的脊髓神经元的影响。 方法:选取新生7d的大鼠,无菌条件下分离骨髓间充质干细胞。选取新生24h的大鼠,无菌条件下分离脊髓背根神经节神经元,并制备机械损伤的模型细胞。用Transwell 可渗透培养皿构建共培养体系,通过细胞形态学,免疫细胞化学方法进行细胞纯度鉴定,观察不同条件作用下神经元特异性烯醇化酶(NSE)阳性细胞的存活。 结果:共培养组神经元细胞NSE阳性率较相应对照组高。在BMSCs与神经元共培养到第6天,计数神经元的存活率,单纯神经元细胞培养组仅有40%的神经元存活,而共培养组神经元的存活率达到65%。单纯损伤神经元细胞培养组仅有26%的神经元存活,共培养组神经元的存活达到51%,细胞凋亡的数量较对照组明显低。 结论:BMSCs可促进正常的脊髓神经元存活,对损伤的脊髓神经元有保护作用,并能诱导神经前体细胞分化为脊髓神经元。
关键词:骨髓间充质干细胞  脊髓损伤  神经元
Effect of bone mesenchymal stem cells on survival of spinal cord neurons after injury using co-culture system    Download Fulltext
Teaching and Research Section of Toxicology,College of Public Health,Jilin University, Changchun,130021
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Abstract:
      Abstract Objective: To explore the effect of bone mesenchymal stem cells(BMSCs) on spinal cord neurons survival and growth using co-culture system. Method: The BMSCs were obtained from P7 rats and spinal cord neurons were obtained from post-born 24h of rats and afterward the neurons were injuried by two edge sword stroke. Experiment set up a co-culture system using Transwell permeable supports. Identification and purification of cells were performed by cell morphology and immunocytochemistry assay, and the survival of neuron-specific enolase(NSE) positive cells were observed. Result: In the group of co-cultured with BMSCs and normal spinal cord neurons, the positive rate of NSE was higher than neurons cultured alone. Only 40% of neurons were seen in cultured alone group, but 65% of neurons were observed in co-cultured group on the 6th d after cultured. In the group of co-cultured with BMSCs and injuried spinal cord nurons, Only 26% of neurons were counted in cultured alone group, and 51% of neurons appeared in co-cultured group. Apoptosis of neurons decreased significantly in co-cultured compared with injuried neurons cultured alone. Conclusion: BMSCs could promote the survival of normal neurons, and protect against neural injury and induce neural differentiation.
Keywords:bone mesenchymal stem cells  spinal cord injury  neuron
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