摘要目的 探讨模拟脊髓细胞环境下骨髓基质干细胞(BMSCs)的分化及蛋白差异表达.方法 分离培养Wistar大鼠骨髓的BMSCs和脊髓的神经细胞,设立BMSCs自然分化组、与神经细胞共培养组和双层培养组,8 d后对各组BMSCs进行神经特异性烯醇化酶(NSE)和胶质纤维酸性蛋白(GFAP)免疫荧光检测.应用表面增强激光解析离子化-飞行时间质谱技术(SELDI-TOF-MS)筛选双层培养组BMSCs变化明显的蛋白进行分析.结果 BMSCs与神经细胞共培养和双层培养8 d后,BMSCs呈神经细胞形态.NSE和GFAP检测结果 ,BMSCs与神经细胞共培养组明显高于BMSCs与神经细胞双层培养组(P＜0.05),而BMSCs和神经细胞双层培养组又明显高于BMSCs自然分化组(P＜0.05).SELDI-TOF-MS检测到TIP39_RAT和CALC-RAT增加到原来的5.360和2.807倍,INSL6-RAT、PNOC_RAT和PCSK1_RAT减少到原来的38.0%、49.9%和43.8%.结论 在脊髓神经细胞微环境下体外培养BMSCs能诱导其分化成神经细胞,而且接触培养分化率高;BMSCs在向神经细胞分化机制中与TIP39_RAT、CALC_RAT、INSL6_RAT、PNOC_RAT和PCSK1_RAT密切相关.

abstractsObjective To explore the differentiation and the variant expression of protein of the bone marrow stromal stem cells(BMSCs)when the BMSCs differentiated into the neuronal cells in the analogous micro-environment of spinal cord injury.Methods BMSCs were isolated from bone marrow of Wister rats and labeled with PKH26(control group),and then were cocultured with neural cells,which were isolated from the spinal cord of the fetal rats,in the same plate well(co-culture group)or in the two1ayer Petri well(two-layer group).Eight days later,the BMSCs were identified by immunotluoreseence staining of NSE and GFAP respectively.The apparently changing proteins were analyzed by SELDI-TOF-MS while the BMSCs differentiated into neurons.Results Eight days after co-culturing with neural cells in the same plate well or in the two-layer Petri well.BMSCs appeared more similar with neural cells.The immunoffuorescence identification showed that.NSE and GFAP of which the BMSCs of the two-layer group expressed were obviously higher than control group(P＜0.05);and these two proteins of co-culture group were also obviously higher than the other two groups(P＜0.05).Five proteins in the co-culture group changed obviously as followed:TIP39_RAT and CALC_RAT were 5.360 and 2.807 times of that in the control group;INSL6_RAT,PNOC_RAT and PCSK1_RAT were 38.0.49.9 and 43.8 percents of these in the control group.Conclusions BMSCs could differentiate into neural cells in vitro,and the differentiation ratio of BMSCs in the co-culture group is higher than that of the two-layer group.Five proteins,including TTP39_RAT,CALC-RAT,INSL6_RAT,PNOC_RAT and PCSK1_RAT,are correlated closely to the mechanisms of which the BMSC8 differentiated into neurons.