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目的建立小鼠胚胎间充质干细胞系C3H/10T1/2细胞成肌、成脂、成内皮、成神经元分化的细胞实验模型。方法用化学诱导剂5-氮杂胞苷诱导C3H/10T1/2细胞成肌、成脂,血管内皮生长因子(vascular endothelial growth factor,VEGF)和碱性成纤维细胞生长因子(basic fibroblast growth factor,bFGF)联合诱导C3H/10T1/2细胞成内皮,bFGF、β-巯基乙醇(β-mercaptoethanol,β-ME)、DMSO诱导C3H/10T1/2细胞成神经元细胞分化。诱导期间应用细胞形态学观察、油红“O”染色、免疫细胞化学染色检测内皮细胞表面标志CD31和神经元特异性烯醇化酶(neuron-specific enolase,NSE)对分化细胞进行鉴定。结果5-氮杂胞苷诱导C3H/10T1/210d后细胞变长,17d后可见明显肌管形成,15d少量细胞出现脂滴,25d油红“O”染色见细胞质大量红色脂滴;VEGF和bFGF诱导5d后细胞呈现“鹅卵石”样形态,8d后阳性表达CD31;bFGF、β-巯基乙醇和DMSO联合诱导3d后细胞胞体收缩,突起变长,15dNSE染色阳性。结论C3H/10T1/2细胞具有多向分化的潜能,可用作研究间充质干细胞生物学特性的模型。
OBJECTIVE: To establish a cell model of myoblasts, adiponectin, endothelium and neuroblast differentiation of mouse embryonic mesenchymal stem cell line C3H / 10T1 / 2. Methods 5-azacytidine was used to induce myogenic, adipogenic, vascular endothelial growth factor (VEGF) and basic fibroblast growth factor (C3F) in C3H / 10T1 / bFGF) induced the differentiation of C3H / 10T1 / 2 cells into neurons by endothelium, bFGF, β-mercaptoethanol and DMSO. During the induction, differentiated cells were identified by morphological observation, oil red staining and immunocytochemical staining for CD31 and neuron-specific enolase (NSE). Results After 5-Azacytosine treatment, C3H / 10T1 / 210d cells became longer and showed obvious myotube formation after 17 days. A small amount of lipid droplets appeared on the 15th day and a large number of red lipid droplets appeared on the cytoplasm after 25 days of oil red staining. After 5 days, bFGF, β-mercaptoethanol and DMSO combined with bFGF, β-mercaptoethanol and DMSO induced the somatic cell contraction, the protuberances became longer and the positive staining of 15dNSE was observed. Conclusion C3H / 10T1 / 2 cells have the potential of multidirectional differentiation and can be used as a model for studying the biological characteristics of mesenchymal stem cells.