在基质材料中生长的许旺细胞:存活、黏附、三维生长和迁移特性  

Survival, adhesion, three-dimensional growth and migration of Schwann cells on biomaterials

作　　者：王光林[1] 裴福兴[1] 杨志明[1] 林卫[2] 毕建红[1] 高伟强[1] 

机构地区：[1]四川大学华西医院骨科,四川省成都市610041 [2]四川大学华西二医院妇产科,四川省成都市610041

出　　处：《中国组织工程研究与临床康复》2007年第40期8038-8042,共5页Journal of Clinical Rehabilitative Tissue Engineering Research

基　　金：国家自然科学基金(30100190);四川省卫生厅基金(000028)~~

摘　　要：目的:将许旺细胞与细胞外基质凝胶、聚乳酸-羟基乙酸共聚物纤维丝复合培养,观察许旺细胞的存活、黏附、三维生长和迁移等生物学行为。方法:实验于2005-02/2006-03在四川大学完成。①实验材料:3～6月龄SD大鼠40只。16～24周流产胎儿由四川大学华西二医院计划生育科提供,产妇及其家属均签署知情同意书,实验经医学伦理委员会批准。聚乳酸-羟基乙酸共聚物(中国科学院成都分院化学所提供);细胞外基质凝胶(Sigma)。②实验方法:将聚乳酸-羟基乙酸共聚物溶解,旋转抽丝法制成直径为20～40μm的聚乳酸-羟基乙酸共聚物纤维丝。无菌条件下取胎儿坐骨神经,去除神经外膜,胰酶消化+差速贴壁法去除成纤维细胞,达80%融合时行BrdU标记。③实验评估:将许旺细胞离心制成0.1～0.2mm3的微粒,接种在细胞外基质凝胶中,观察许旺细胞在凝胶中的存活、生长和移行。将许旺细胞制成1×1010L-1的30%细胞外基质凝胶后,分别滴加在经过胶原、细胞外基质凝胶预处理的聚乳酸-羟基乙酸共聚物纤维丝的一端,观察许旺细胞在纤维丝上的黏附、生长和迁移。将BrdU标记的许旺细胞复合细胞外基质凝胶后,置入聚乳酸纤维管用以修复大鼠坐骨神经缺损,术后3,6周取材,BrdU免疫组化染色观察许旺细胞的存活情况。结果:①许旺细胞在无血清状态下能在细胞外基质凝胶中存活,并分裂、增殖,相互移行形成类似Bungner带状的许旺细胞柱状结构。②许旺细胞能贴附在聚乳酸-羟基乙酸共聚物纤维丝上生长并沿纤维丝移行,形成多层排列的类似Bungner带状的许旺细胞柱,细胞外基质凝胶能显著增加贴附和移行细胞数量,而胶原作用不显著。③许旺细胞和细胞外基质凝胶复合移植,6周后BrdU阳性细胞数明显低于第3周(t=3.71,P<0.01),成活细胞能在细胞外基质凝胶中移行。结论:许旺细胞与细胞外基质凝胶、聚乳AIM： To study the biological behaviors （survival, adhesion, three-dimensional growth and migration） of Schwann cells in extracellular matrix （ECM） gel and on poly（lactic-co-glycolic acid） （PLGA） fibers. METHODS： The experiment was carried out in Sichuan University from January 2005 to March 2006.①Materials：. Forty SD rats aged 3-4 months. And 16-24 weeks fetal samples were supplied by Department of Planned Parenthood in West China Second Hospital to Sichuan University, with the informed consents of puerperants and their families. PLGA was offered by Chengdu Branch of Chinese Academy of Sciences; ECM gel （Sigma）. ②Degraded PLGA polypore biomaterials was used to form complex PLGA fibers （20-40 μm diameter） using rotation silking method. Schwann cells were obtained from fetal sciatic nerves sterilely, followed by the removals of epineurium and fibroblasts using the techniques of trypsinization and differential adhesion, then the rate of BrdU-labeled Schwann cell was above 80%.③ Schwann cells particulates （0.1 mm^3-0.2 mm^3） were seeded into ECM gel to observe the survival, growth and migration of Schwann cells. BrdU-labeled Schwann cells were seeded into 30% ECM/DMEM gel at 1 ×10^10 L^-1, then implanted on one end of PLGA fibers which were previously treated with collagen or ECM gel, and were co-cultured with PLGA fibers. The adhesion, growth and migration of Schwann cells on the PLGA fibers were observed. BrdU-labeled Schwann cells were seeded in ECM gel and implanted into PLA conduits to repair the defects of rat sciatic nerve. The survival of Schwann cells was detected by BrdU immunohistochemical method at the 3^nd and 6^th weeks after operation. RESULTS：① Schwann cells survived in ECM gel, and could proliferate and migrate to form Bungner band-like structures of Schwann cell columns without plasma.②Schwann cells could adhere and migrate on PLGA fibers, and these abilities could remarkably promote by ECM gel other than collagen. ③When seeded into ECM gel, most of

关 键 词：许旺细胞 组织工程 细胞外基质凝胶 神经损伤 生物材料 

分 类 号：R318.08[医药卫生—生物医学工程]