摘要
目的探讨人脐血间充质干细胞用于神经细胞再生的可行性与人脐血间充质干/祖细胞最佳的诱导培养条件。寻找一种合适的细胞来源进行移植以代替受损的神经元、星形胶质细胞和少突胶质细胞来修复神经损伤。方法将脐血单个核细胞置于低血清(2%)达氏修正依氏培养基中培养,生成贴壁细胞层,依传代方法,用相同培养条件进行扩增,扩增后的贴壁细胞置入细胞培养液(加入新生大鼠神经细胞分离培养上清液)中,并添加表皮生长因子(EGF)和碱性成纤维细胞生长因子(bFGF)进行诱导分化。采用流式细胞术、免疫组织化学法分别检测被诱导的神经样细胞的表面标志及细胞特异性标志、结构,并进行分析。结果脐血间充质干/祖细胞克隆在脐血单个核细胞中出现频率为0.5×10-6,在传20代时,可有效扩增1.5×106倍,诱导后,70%的脐血间充质干/祖细胞分化为神经样细胞。结论采用上述扩增与诱导条件,脐血间充质干/祖细胞可得到有效扩增,并可高效向神经样细胞分化,从而证实脐血是一种细胞替代治疗前景光明的细胞来源。
Objective To investigate the feasibility of inducing human cord blood mesenchymal stem cell into nerve cell, to explore the optimal culture condition of inducing cord blood mesenchymal stem and progenitor cell,so as to provide transplant with a kind of cell source to replace impaired neuron,astrocyte and oligodendrocyte for repairing nerve injury. Methods Put mononuclear cells of cord blood into low serum(2%)DMEM substratum and cultivate them into adherent cells. According to passage method, amplifying them under the same cultivating condition. Put the amplified adherent cells into cell culture fluid(adding the supernatant of nerve cell fractional culture of neonate rats) and add epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF) to induce cell differentiation. Flow cytometry, immunohistochemistry and histochemistry were used to detect and analyze the surface marker, specific marker and structure of induced neuroid cells. Results The occurrence rate of cord blood mesenchymal stem and progenitor cell clone in mononuclear cells of cord blood was 0.5×10-6. After passaging 20 generations, it could be amplified by 1.5×106 times;through induction, 70% of cord blood mesenchymal stem and progenitor cells were differentiated into neuroid cells. Conclusion Following the above-mentioned amplification and inducing conditions, cord blood mesenchymal stem and progenitor cells can be amplified and differentiated into nerve -like cells efficiently. It is proved that cord blood is a kind of promising cell source in cell replacing treatment.
出处
《中国药物与临床》
CAS
2005年第7期516-519,共4页
Chinese Remedies & Clinics
基金
山西医科大学学生创新基金资助项目(2004070)
