GDNF转染雪旺氏细胞构建神经导管复合体

Construction of nerve conduits with Schwann cells transfected by GDNF genes on PLGA

目的利用GDNF(胶质神经源性神经营养因子)基因转染的雪旺氏细胞和PLGA(聚乳酸/聚乙醇酸共聚物)管共同培养构建神经导管复合体。方法利用乳鼠臂丛神经和坐骨神经培养雪旺氏细胞,再把pcDNA3.1(+)/GDNF质粒通过脂质体转入细胞内。利用PCR检测转染雪旺氏细胞的GDNF表达。将转染雪旺氏细胞扩增达到一定数量后,用微注射和共培养法,与PLGA管构建神经导管。通过光镜和扫描电镜检测雪旺氏细胞的生长情况。结果成功培养出雪旺氏细胞,并转入GDNF基因。转染的雪旺氏细胞可表达GDNF,PCR显示转染的雪旺氏细胞GDNF表达较正常细胞明显提高。转染的雪旺氏细胞可在PLGA管表面贴壁、生长。结论GDNF基因转染的雪旺氏细胞可与PLGA管共同构建神经导管复合体。

Objective To obtain nerve conduits with Schwann cells transfected by GDNF genes and PLGA. Methods Schwann cells were harvested from the rat brachial plexus and sciatic nerves. After the Schwann cells were transfected by pcDNA3.1（＋）/GDNF, the express of GDNF was assessed through PCR. When Schwann cells proliferated to 2-5× 107 Schwann cells /ml, then they were injected into the PLGA conduits. We investigated growth conditions of Schwann cells with light microscopy and scanning electron light microscopy. Results Schwann cells were harvested and transfected by pcDNA3.1（＋）/GDNF successfully. GDNF expressed in Schwann cells. Schwann cells attached to and grew well on the PLGA conduits. Conclusion The nerve conduits could be constructed by seeding Schwann cells transfected by GDNF genes within the PLGA conduits.