大鼠脂肪干细胞向雪旺细胞诱导分化能力的研究  

Study on differentiation of rat adipose tissue-derived stromal cells into Schwann-like cells

作　　者：任志午[1,2] 赵喆[1] 王玉[1] 陈继凤[1] 詹胜锋[1] 刘炎[1] 许文静[1] 张莉[1] 彭江[1] 卢世璧[1] 

机构地区：[1]解放军总医院骨科研究所,北京100853 [2]南开大学医学院,天津300071

出　　处：《中国应用生理学杂志》2011年第4期385-388,I0012,共5页Chinese Journal of Applied Physiology

基　　金：国家863计划(2009AA03Z312);全军十一五专项项目(06Z057);国家支撑计划(2009BAI87B02)

摘　　要：目的:研究脂肪干细胞(ADSCs)向雪旺细胞的诱导分化,为神经组织工程提供新的种子细胞。方法:取SD大鼠项背处的皮下脂肪,分离出脂肪干细胞并培养传代,流式细胞仪检测细胞表面特异标记CD29,CD34,CD44,CD45,CD90,以评价干细胞的生物学特性;采用b-FGF和forskolin等诱导脂肪干细胞向雪旺细胞分化,光镜观察诱导后细胞形态的变化;免疫荧光染色鉴定雪旺细胞特异性标记物S100、P75和GFAP的表达;PCR检测诱导前后雪旺细胞特异性标记物S100、P75的表达。结果:分离培养的鼠脂肪干细胞CD29、CD90表达呈阳性,而CD34、CD44和CD45表达呈阴性,具有脂肪干细胞的生物学特性;脂肪干细胞经过胶质细胞生长因子的作用,光镜下发现诱导的细胞形态与雪旺细胞相似;免疫荧光染色S100、P75和GFAP阳性;RT-PCR结果显示诱导的雪旺细胞标记物S100和P75表达上调。结论:脂肪干细胞可诱导分化成雪旺细胞,其表型和分子特征与雪旺细胞相似,诱导分化的脂肪干细胞是一种理想的神经组织工程的种子细胞。Objective： To investigate the phenotypic, molecular and biological characteristics of adipose tissue-derived stromal cells （ADSCs） differentiated alonely a Schwann cells（SCs） lineage and to provide a new cells＇ seed source for nerve tissue engineering or cell therapy. Methods： Cultured ADSCs were isolated from SD rats and the undifferentiated ADSCs were confirmed by detection of MSC-specific cell-surface markers. The ADSCs were differentiated along a glial cell lineage using an established cocktail of growth factors. Following differention, we used immunofluorescene staining and RT-PCR to evaluate the characteristics of differentiated WJMSCs. Results： ADSCs were successfully isolated from the rats＇ fat tissue. The isolated ADSCs expressed CD29, CD90 but not CD34, CD44 nor CD45. Osteogenic differentiation was detected by Alizarin red staining and adipogenic differentiation was comfirmed by Oil-red O staining. ADSCs treated with a mixture of glial growth factors adopted a spindle-like morphology similar to Schwann cells. Immunocytochemical staining and RT-PCR analysis revealed that the treated cells expressed the glial markers S100, P75 and glial fibrillary acidic protein indicative of differentiation. Conclusion： ADSCs can be differentiated into cells that are Schwarm-like in terms of morphologic features and phenotype and could be suitable Schwann-cell substitutes for nerve repair in clinical applications.

关 键 词：脂肪干细胞 雪旺细胞 诱导分化 

分 类 号：Q813.1[生物学—生物工程]