乳鼠肌源性干细胞向许旺样细胞的诱导分化  

In vitro induced differentiation of mouse muscle-derived stem cells into Schwann-like cells

作　　者：宋艳玲[1,2] 唐乙[1,2] 丁维进[1,2] 王春燕[2] 苏志达[2] 江华[1] 张盈帆[1] 

机构地区：[1]解放军第二军医大学长征医院整形外科,上海市200003 [2]解放军第二军医大学长征医院神经科学研究中心,上海市200003

出　　处：《中国组织工程研究》2013年第10期1841-1848,共8页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金(青年科学基金项目)项目(81100950);上海市科委基础研究重点课题(08JC1407100)~~

摘　　要：背景:有证据显示,肌源性干细胞能够促进肌纤维的再生,增强再生肌组织的功能,并具有分化为神经外膜组织细胞和具有许旺细胞表型细胞的潜能。目的:分析许旺细胞条件培液诱导小鼠乳鼠源肌源性干细胞向许旺细胞的分化,探讨其作为神经组织工程的种子细胞的可行性。方法:取C57BL/6乳鼠四肢骨骼肌,采用改良的Preplate技术纯化培养肌源性干细胞,利用抗体Desmin和Sca-1对纯化后的细胞进行免疫细胞化学鉴定。采用许旺细胞条件培液诱导其向许旺细胞分化,并行许旺细胞特异性抗体S100β、P75NTR免疫细胞化学鉴定。结果与结论:从小鼠骨骼肌中分离得到肌源性干细胞,能在体外稳定增殖传代,Desmin和Sca-1免疫细胞化学染色阳性。肌源性干细胞经体外诱导分化后部分细胞S100β、P75NTR染色阳性。说明应用改良的Preplate方法,可从新生小鼠骨骼肌中分离得到肌源性干细胞;采用许旺细胞条件培液能将其诱导分化为许旺样细胞,有可能成为神经组织工程神经较理想的许旺细胞替代物。BACKGROUND： There is evidence that muscle-derived stem cells can promote the regeneration of muscle fiber, strength the function of regenerative muscle tissue, exhibit the ability to differentiate into epineurial cells and possess the phenotype of Schwann cells. OBJECTIVE： To analyze the induced differentiation of neonate mouse muscle-derived stem cells into Schwann-like cells by Schwann cell conditioned culture medium and investigate the feasibility of use of neonate mouse muscle-derived stem cells as the seed cells for nerve tissue engineering. METHODS： Muscle-derived stem cells were harvested from C57BL/6 neonate mouse appendicular skeletal muscle and purified and cultured by modified Preplate technique. These cells were identified by immunocytochemistry with Desmin and Sca-1 antibodies and then induced to differentiate into Schwann-like cells using Schwann cell conditioned culture medium. Schwann cell specific antibodies S100β and P75 NTR were used for immunocytochemical identification. RESULTS AND CONCLUSION： Muscle-derived stem cells isolated from mouse skeletal muscle could be stably proliferated and passaged, and were positive for Desmin and Sca-1 staining. After induced differentiation, some muscle-derived stem cells were positive for S100β and P75 NTR staining. These findings suggest that muscle-derived stem cells can be harvested from neonate mouse skeletal muscle through the use of modified Preplate method. In the Schwann cell conditioned culture medium, neonate mouse skeletal muscle-derived stem cells can differentiate into Schwann cell-like cells and therefore can become an ideal Schwann cell substitute in nerve tissue engineering.

关 键 词：干细胞 干细胞培养与分化 肌源性干细胞 许旺细胞 许旺样细胞 培养基 诱导分化 骨骼肌 三重酶消化 神经轴突 神经组织工程 种子细胞 国家自然科学基金 干细胞图片文章 

分 类 号：R394.2[医药卫生—医学遗传学]