周围神经浸出液对犬脂肪干细胞向神经膜细胞分化的影响  

Effect of peripheral nerve leachate on Schwann cell differentiation of dog adipose derived stem cells

作　　者：张自强[1] 刘玉梅[1] 位兰[1] 朱雪敏[1] 黄婷婷[1] 张春燕[1] 

机构地区：[1]河南科技大学动物科技学院,河南洛阳471003

出　　处：《中国兽医科学》2015年第7期740-745,共6页Chinese Veterinary Science

基　　金：国家自然科学基金项目(31101779)

摘　　要：本研究旨在研究周围神经浸出液对犬脂肪源性干细胞(adipose derived stem cells,ADSCs)向神经膜细胞(Schwann cell,SCs)分化的影响。采用Ⅰ型胶原酶消化法体外原代培养犬ADSCs,并应用流式细胞术进行表型分析,同时观察所培养细胞的成脂成骨分化能力。分离犬坐骨神经,无菌剪碎、浸出和过滤制备犬周围神经浸出液。将周围神经浸出液作为诱导剂,作用于犬ADSCs,7d后分别通过倒置显微镜观察细胞形态的变化;免疫荧光试验和Western-blot观察SCs标志蛋白GFAP和S-100的表达情况。结果显示,周围神经浸出液作用于犬ADSCs 7d后,大多数细胞形态由原来的三角形或梭形变为两端伸出2个细长突起的长梭形,非常像成熟SCs的形态;免疫荧光和Western-blot试验显示,周围神经浸出液诱导ADSCs后,细胞出现GFAP和S-100阳性表达。以上结果提示,周围神经浸出液能有效诱导犬ADSCs向SCs分化。In order to observe the effect of peripheral nerve leachate on Schwann cells differentiation into dog adipose derived stem cells（ADSCs） ,we cultured dog ADSCs in vitro by digestion with I type col- lagenase, and confirmed phenotype by flow cytometry. At the same time, the ability of adipocyte differentia- tion into osteoblast was assessed. Dog sciatic nerves were isolated, cut, soaked in culture medium, and then filtered as peripheral nerve leachate. ADSCs were incubated with peripheral nerve leachate for 7 days. Then the morphology of cells was observed by inverted microscope,and the protein expressions of S-100, and GFAP proteins were confirmed by immunofluorescence and Western-blot. The results showed that almost all of the treated ADSCs with peripheral nerve leachate displayed a spindle shaped-like morphology with two slender processes similar to that of Schwann cells. Immunocytochemistry and Western-blot confirmed that incubated ADSCs by peripheral nerve leachate were positive for S100 and GFAP. These results suggest that peripheral nerve leachate could induce dog ADSCs differentiation into Schwann cells.

关 键 词：脂肪干细胞 神经膜细胞 周围神经 分化 

分 类 号：S852.21[农业科学—基础兽医学]