羊膜脱细胞基质对脐血间质干细胞突触蛋白表达的影响  

Effect of human acellular amniotic membrane matrix on synapsin expression of umbilical cord blood mesenchymal stem cells

作　　者：胡炜[1,2] 杨枫[1] 唐尤佳 陈希兰[1] 杨波[2,3] 关方霞[4] 

机构地区：[1]九江市第一人民医院神经外科,江西省九江市332000 [2]郑州大学第一附属医院神经外科,河南省郑州市450052 [3]河南省高等学校临床医学重点学科开放实验室,河南省郑州市450052 [4]郑州大学生物工程系,河南省郑州市450001

出　　处：《中国组织工程研究与临床康复》2011年第14期2500-2503,共4页Journal of Clinical Rehabilitative Tissue Engineering Research

摘　　要：背景:天然细胞外基质对成体干细胞神经发育的影响,是目前的研究热点,国内外研究多侧重于神经标志物表达的研究,针对突触发育的研究较少。目的:观察羊膜脱细胞基质对脐血间质干细胞突触素表达的影响。方法:实验组将第 3 代人脐血间质干细胞接种在包被膜样基质盖玻片的 24 孔板中,以接种时开始计时,细胞以基础培养基培养 4 d。对照组无膜样基质,余同实验组。苏木精-伊红染色观察细胞形态变化,免疫组化 SP 法检测接种第 2、4 天细胞突触素相关蛋白表达,RT-PCR 法检测接种前和接种后第 2、4 天细胞突触素相关蛋白Ⅰ、Ⅱ、Ⅲ基因转录水平的表达。结果与结论:对照组呈成纤维细胞形态,实验组细胞突起延伸,相互连接;第 2、4 天实验组突触素相关蛋白表达均明显高于对照组,差异有显著性意义(P < 0.01);接种前及对照组第 2、4 天细胞突触素相关蛋白Ⅰ、Ⅱ、Ⅲ基因转录水平无表达,实验组第 2,4 天细胞突触素相关蛋白Ⅱ基因转录水平低表达,突触素相关蛋白Ⅲ弱表达,突触素相关蛋白Ⅰ不表达。结果提示羊膜脱细胞基质可以促使人脐血间质干细胞突起的产生、延伸并部分相互连接,表达突触素相关蛋白,为脐血间质干细胞神经分化提供了有利的条件。BACKGROUND：The effect of natural extracellular matrix on neural development of adult stem cell is a hot research spot currently.More studies on neural markers expression at home and abroad,few studies on synapse development.OBJECTIVE：To investigate the effect of human acellular amniotic membrane matrix on synaptophysin expression of umbilical cord blood mesenchymal stem cells.METHODS：The 3rd generation of human cord blood mesenchymal stem cells in experimental group was seeded in 24 orifice plate of coating membrane-like matrix cover glass.Time was counted when the inoculation,cells were cultured by basal medium for 4 days.There was no membrane-like matrix,the remaining procedures was the same as the experimental group.Morphologic changes of cells were observed by hematoxylin-eosin staining.Synaptophysin-related protein expression（SYN expression） was detected by immunohistochemical SP method at 2 and 4 days inoculation.SYN expression Ⅰ,Ⅱ,Ⅲ of genetic transcription level was detected by RT-PCR method at 2 and 4 days before and after inoculation.RESULTS AND CONCLUSION：Cells in control group was fibroblast-like,cells in experimental group extended the processes and connected each other.SYN expression in experimental group was higher compared with control group;there were significant differences（P0.01）.SYN Ⅰ,SYN Ⅱ,SYN Ⅲ did not express in transcriptional level both before inoculation and at 2,and 4 days in control group.At 2 and 4 days in experimental group,SYN Ⅱ had low expression,SYN Ⅲ had poor expression,and SYN Ⅰ did not express in transcriptional level.The results indicated that acellular amniotic membrane matrix can promote the production and extension of the umbilical cord blood mesenchymal stem cells process,and connected each other,express SYN,and provide favorable conditions for neural differentiation of umbilical cord blood mesenchymal stem cells.

关 键 词：细胞外基质 脐血间质干细胞 分化 干细胞 突触素相关蛋白 

分 类 号：R394.2[医药卫生—医学遗传学]