猕猴骨髓间充质干细胞与去细胞神经优化人工神经的体外构建  

The consturction of artificial nerves by planting rhesus maceque mesenchymal stem cells into chemically extracted allogenous accellular nerves in vitro

作　　者：杨光诗[1] 许扬滨[1] 朱家恺[1] 刘小林[1] 朱庆棠[1] 

机构地区：[1]中山大学附属第一医院显微外科

出　　处：《中华显微外科杂志》2005年第1期35-37,i002,共4页Chinese Journal of Microsurgery

基　　金：广州市科技攻关重大项目(2002Z1 E0031)

摘　　要：目的　探讨猕猴骨髓间充质干细胞作为种子细胞与去细胞同种异体神经作为支架,体外构建人工神经的研究。　方法　采用密度梯度离心的方法分离并培养猕猴骨髓间充质干细胞,利用相差显微镜观察细胞生长情况并描绘其生长曲线;取猕猴3段4 cm的周围神经,其中2段用4%triton X 100及4%脱氧胆酸钠液分别萃取1次和2次,在新鲜神经和萃取神经中段取材,组织学检测及电镜学观察萃取前后的形态学变化;采用显微注射的方法把猕猴骨髓间充质干细胞种植到萃取两次的异体神经内,HE染色观察细胞在支架内的生长情况。　结果　猕猴骨髓间充质干细胞自我增殖能力强;萃取1次的异体神经细胞及髓鞘结构仍部分存留,而萃取2次的其细胞及髓鞘结构消失;HE染色可见细胞在萃取2次的异体神经内可爬行生长。　结论　猕猴骨髓间充质干细胞与化学萃取的去细胞同种异体神经适合体外构建人工神经。Objective To take rhesus maceque mesenchymal stem cells(MSCs) as seed cells and take chemically -extracted allogenous accellular nerve grafts(ANGs) as frames, and artificial neryes were constructed in vitro. Methods Through density gradient centrifugation rhesus maceque mesenchymal stem cells were got and observed under the phase contrast microscope. The growth curve of MSCs was described. ANGs were got by extraction of 4% triton -X -100 and 4% sodium deoxycholate and were detected in histology and electron microscopy to learn the morphologcal changes. Microinject the mesenchymal stem cells into the ANGs and study the cells migration in the artificial nerves in vitro by HE staining. Results Rhesus maceuqe mesenchymal stem cells can be got in mass by density gradient centrifugation. In the one -time extracted ANGs parts of myelin and Schwann cells still remained but there was no myelin and Schwann cells in two -time extracted ANGs. HE stain show MSCs could grow well and migrate in ANGs. Conclusion Rhesus macaque mesenchymal stem cells and allogenous accellular nerve grafts extracted chemically are suitable to construct artificial nerves in vitro

关 键 词：骨髓间充质干细胞 去细胞 体外 猕猴 人工神经 同种异体 HE染色 密度梯度离心 显微注射 生长曲线 

分 类 号：R318.1[医药卫生—生物医学工程]