去细胞神经种植脂肪干细胞的体外培养

An experimental study on implanting adipose tlssue-derived stromal cells into acellular nerve in vitro

目的探讨大鼠脂肪干细胞（ADSCs）与去细胞神经体外结合培养的方法。方法从成年F344大鼠的脂肪组织中分离培养ADSCs，用携带绿色荧光蛋白基因的复制缺陷性重组腺病毒载体（Ad-GFP）转染ADSCs。采用Triton X-100和脱氧胆酸钠去除大鼠坐骨神经中的细胞成分，将转染GFP的ADSCs通过微注射方法注入去细胞神经。用HE染色、电镜、荧光显微镜观察神经萃取及细胞种植后ADSCs的生长情况。结果Ad—GFP转染后，约85％的ADSCs可激发出绿色荧光。化学萃取可有效去除神经纤维中的细胞，ADSCs可在去细胞神经内生长并迁移。结论ADSCs通过微注射方法注入去细胞神经而构建的复合体可能成为一种新的神经缺损修复材料。

Objective To explore a feasible way rat adipose tissue-derived stromal cells （ADSCs） can be implanted into acellular nerve in vitro. Methods ADSCs were isolated from the adipose tissue of adult F344 rats, and expanded in culture medium. The cultured ADSCs were transfected with replication-deficient recombinant adenovirus vectors containing green fluorescent protein gene （Ad-GFP） . The rat sciatic nerves were extracted with Triton X-100 and sodium deoxycholate by removing cells. The ADSCs transfected with GFP were micro-injected into the acellular nerves. HE staining, electron microscopo and fluorescent microscopy were used to observe the growth of the cells. Results After ADSCs were transfected with Ad-GFP, green fluorescence was triggered in approximately 85% of ADSCs. The cells in the nerves could be effectively removed by chemical extraction. ADSCs could survive and migrate in the acellular nerves in vitro. Conclusion Acellular nerves populated with ADSCs may be a new kind of material to repair nerve defects after injury.