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目的:探讨神经细胞形成的体外微环境,诱导骨髓基质干细胞(bonemarrowstromalcell,BMSC)向神经元分化的机制。方法:从SD大鼠骨髓中提取BMSC进行体外培养、扩增,并用免疫组化染色进行鉴定。以绿色荧光染料PKH67标记BMSC后,将BMSC与神经细胞共培养以及用双层培养皿联合培养8d后,用免疫荧光检测BMSC是否分化为神经元。结果:将BMSC与神经细胞共培养后,BMSC出现神经元的形态特点,且有(32.72±2.56)%的神经元表达特异性烯醇化酶(neuron specificenolase,NSE),与BMSC自然分化组(对照组)相比较差异非常显著(P<0.05);但用双层培养皿联合培养时,只有(4.87±0.79)%的BMSC表达NSE,与对照组相比较差异不显著(P>0.05),但与BMSC和神经细胞共培养组相比较差异显著(P<0.05)。结论:体外培养时,神经细胞形成的局部微环境可促进BMSC向神经元方向分化,并且细胞间的紧密接触是诱导BMSC向神经元分化的重要条件。
Objective: To investigate the in vitro microenvironment formed by nerve cells and to induce the differentiation of bone marrow stromal cells (BMSCs) into neurons. Methods: BMSCs were extracted from the bone marrow of SD rats for in vitro culture, expanded, and identified by immunohistochemical staining. After BMSCs were labeled with green fluorescent dye PKH67, BMSCs were co-cultured with nerve cells and cultured with double-layer Petri dishes for 8 days. Then, BMSCs were differentiated into neurons by immunofluorescence. RESULTS: BMSC showed morphological characteristics of neurons after co-culture with BMSCs, and there were (32.72 ± 2.56)% neurons specific enolase (NSE) expressing neurons in BMSCs. Compared with BMSCs naturally differentiated group (4.87 ± 0.79)% of BMSCs expressed NSE, but the difference was not significant (P> 0.05) compared with that of control group (P> 0.05). However, Compared with BMSC and neural cell co-culture group, the difference was significant (P <0.05). CONCLUSION: Local microenvironment formed by nerve cells can promote the differentiation of BMSC into neurons in vitro, and the close contact between cells is an important condition for inducing BMSCs to differentiate into neurons.