永生化施万细胞和神经细胞建立体外共培养模型及神经细胞对髓鞘特异性基因表达的影响  

The establishment of co-culture model in vitro with Schwann cell and nerve cell and the effect of nerve cell on the expression of myelin-specific genes

作　　者：牛群丽[1] 钟延丰[1] 由江峰[1] 王盛兰[1] 郑丹枫[1] 张燕[1] 

机构地区：[1]北京大学医学部基础医学院病理学系,北京市100191

出　　处：《河北医药》2011年第7期965-968,共4页Hebei Medical Journal

基　　金：国家自然科学基金项目(编号:30670794)

摘　　要：目的通过体外共培养永生化施万细胞株RSC96细胞和神经细胞株VSC4.1细胞,建立施万-神经细胞共培养模型,观察髓鞘化过程中神经细胞对细胞增殖和髓鞘特异性基因MBP、PMP22表达的影响。方法建立施万细胞和神经细胞共培养模型:本研究选用RSC96和VSC4.1细胞进行共同培养。实验分为3组:单独培养的RSC96细胞组(RSC96组)、VSC4.1细胞组(VSC4.1组)、共培养组。免疫荧光染色对细胞进行鉴定。经光镜及电子显微镜观察,确定共培养的两种细胞可发生相互作用。利用MTT法检测细胞增殖情况。RT-PCR方法检测髓鞘特异性基因MBP、PMP22的表达。结果 RSC96细胞包绕VSC4.1细胞的轴突形成髓鞘。MTT示:RSC96细胞增殖速度明显快于VSC4.1细胞。但当两种细胞共培养时,细胞的增殖速度与单独的RSC96细胞相比,明显下降。在单独培养的RSC96细胞中,PMP22基因的表达有先增高后降低的现象。在共培养模型中,PMP22基因的表达是逐渐增高的。在前2d的表达量低于单独培养的RSC96细胞。单独培养的RSC96细胞及共培养组细胞在前3d均无MBP的表达。结论永生化施万细胞株RSC96细胞和神经细胞株VSC4.1细胞可以建立施万-神经细胞共培养模型。神经细胞可以调控施万细胞的增殖速度与髓鞘特异性基因的表达模式。Objective To establish the Schwann cell （SC） and nerve cell （NC） co-culture model by using immortalized RSC96 cell and ventral spinal cord4.1 cell （VSC4.1）,and to observe the effects of nerve cell on the proliferation and expression of myelin-specific genes-MBP,PMP22 during the process of co-culture.Methods The Schwann-nerve cells co-culture model was established by using hybrid motorneuronal cell line （VSC4.1） and immortalized rat Schwann cell （RSC96）.There were three groups：single-RSC96 groups,single-VSC4.1 groups and co-cultured groups.The RSC96 cells and VSC4.1 cell were identified by immunofluorescence method.The phase contrast microscopy and electron microscopy were used to observe the interaction of the co-cultured cells.The cell proliferation and the expression of MBP and PMP22 were detected by MTT and RT-PCR.Results The cell membrane of RSC96 could wrap the axon of VSC4.1 cell to form myelination.The MTT examination showed that the proliferation of RSC96 cells was significantly faster than that of VSC4.1 cells,however,when RSC96 cells were co-cultured with VSC4.1 cells,the cell proliferation was slower than that of single-RSC96.The gene expression of PMP22 in single cultured RSC96 cells was increased,then decreased on the third day.In the co-culture model,the expression of PMP22 gene was increased gradually,but it was less than that of the single cultured RSC96 cells on the first two days.The expression of MBP was not found during the three days.Conclusion RSC96 and VS4.1 cells can be used to establish the co-cultured model.NC can regulate SC proliferation and myelin-specific gene expression.

关 键 词：RSC96细胞 VSC4.1细胞 共培养 髓鞘特异性基因 

分 类 号：R322.85[医药卫生—人体解剖和组织胚胎学]