骨髓基质细胞在传代与诱导分化为神经细胞过程中hes1和mash1基因的表达变化

Expression changes of hes1 and mash1 gene during the bone marrow stromal cells serially passaging and differentiating into neural cells

目的:探讨骨髓基质细胞(BMSCs)在传代培养与诱导分化为神经细胞过程中hes1和mash1基因的表达变化及作用。方法:采用全骨髓培养法体外分离培养获得BMSCs,倒置显微镜下观察其形态变化,应用半定量RT-PCR技术分析hes1和mash1基因在传代和诱导分化为神经细胞过程中的动态时序表达。结果:诱导后细胞形态出现神经元样改变。hes1在第3代BMSCs诱导后(P)第1日表达下降,之后第3日表达增加。mash1在BMSCs传至第5代时高表达,随后急剧下降;P3的BMSCs诱导后表达逐日增加。结论:hes1和mash1基因在BMSCs传代与诱导分化为神经细胞过程中可能起重要作用。

Objective： To detect the expression changes and effects of hes1 and mash1 gene during the bone marrow stromal cells （BMSCs） serially passaging and differentiating into neural cells. Methods： BMSCs isolated from bone marrow were cultured in vitro. Then BMSCs of passage 3 were induced into neural cells by treated with DMEM/F12, B27, bFGF and RA. The morphologic features and growth status of BMSCs and differentiated BMSCs were observed by invert microscope. Semi-quantitative reverse transcription polymerase chain reaction （RT-PCR） was used to detect the expressions of hesl and mash1 in mRNA level in the BMSCs of P3, 5, 7, 9 and P3 BMSCs after induction for 1 and 3 d. Results：The neuron-like cells derived from P3 BMSCs were observed after induction. The expression of hesl mR- NA was decreased in a significant and progressive manner as serially passaging; it had a temporal decrease at 1 d and then a significant increase at 3 d for induced P3 BMSCs. The mash1 mRNA levels significantly increased in P5 BMSCs and then deceased in P7 and P9 BMSCs; it had a progressive increase for P3 BMSCs after induction. Conclusion： The hesl and mash1 gene may play an important role during the bone marrow stromal cells serially passaging and differentiating into neural cells.