脐带间充质干细胞诱导分化为类许旺细胞的实验研究  

Differentiation of Human Umbilical Cord Mesenchymal Along a Schwann Cells Lineage

作　　者：祝加学[1] 陆雄伟[1] 沈尊理[2] 秦金保[3] 王洋[2] 金晨[1] 

机构地区：[1]上海交通大学医学院附属第三人民医院骨科,上海201999 [2]上海交通大学附属第一人民医院创伤中心,上海200080 [3]上海交通大学医学院附属第九人民医院血管外科,上海200111

出　　处：《现代生物医学进展》2015年第5期834-838,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目(30872630)

摘　　要：目的:探讨通过化学诱导的方法,诱导脐带间充质干细胞分化为类许旺细胞,为组织工程神经寻找一种新的种子细胞来源。方法:通过酶消化法,分离获得原代脐带间充质干细胞。体外培养至第3代(P3),以1×103/cm2接种于培养瓶中,待细胞长至亚融合状态,吸弃培养液,加入含β-巯基乙醇的培养液预诱导24 h。然后,加入含有全反式维甲酸的培养液进一步诱导72 h。最后,加入含有胶质细胞生长因子的培养液,作用两周。对诱导后的脐带间充质干细胞用免疫荧光及RT-PCR法,进行形态观察和表型鉴定。结果:经过上述诱导过程,脐带间充质干细胞的形态,由开始时的扁平、片状、多极,类似成纤维细胞状,逐渐变为长梭形,双极或三极,类似许旺细胞。同时,表达许旺细胞特异性标志S-100、P75;而且S100,P75的m RNA水平也明显上调,并且可以观测到GFAP的m RNA条带。结论:化学诱导法,可以将脐带间充质干细胞诱导分化为类许旺细胞,有望为组织工程神经提供一种新的许旺细胞来源。Objective： To explore the potential differentiation of Schwann cells from human umbilical cord mesenchymal stem cells under the induction of chemical agents, which may provide a new source of seed cells for neural tissue engineering. Methods： The primary human umbilical cord mesenchymal stem cells were harvested by digestion and isolation. After the 3rd passage, the cells were plated at 1 ×10^3/cm2 in the flask and treated with β-mercaptoethanol for 24h when the cells were subconfluent. Afterwards, the cells were cultivated with all-trans-retinoic acid for 72h and ttien incubated with glial growth factors for two weeks. Then the morphology and phe- notype of induced umbilical cord mesenchymal stem cells were identified through immunocytochemical staining and RT-PCR. Results： The primary umbilical cord tnesenchymal stem Cells were flat, platy, multipolar like Schwann cells, but shifted to spindle shape, bipolar or tripolar like fibroblast cells, Moreover, the specific markers for Schwann cells, S100 and P75, were positively expressed in the induced cells. The mRNA levels of S100 and P75 also significantly increased along with the expression GFAP. Conclusion： Schwann-like cells can be induced from umbilical cord mesenchymal stem cells by induction with chemical agents, which has the potential to be a novel cell source for neural tissue engineering.

关 键 词：脐带间充质干细胞 类许旺细胞 诱导 胶质细胞生长因子 

分 类 号：Q813[生物学—生物工程] R318[医药卫生—生物医学工程]