犬骨髓间充质干细胞体外培养及诱导成软骨特性初步研究  

Effect of age and origin on proliferation and chondrogenic differentiation capabilities of canine bone marrow mesenchymal stem cells in vitro

作　　者：李尚滨[1] 霍然[1] 于庆平[1] 张文浩[1] 高峰[1] 赵阳[1] 

机构地区：[1]山东大学山东省立医院美容整形烧伤科,山东济南250021

出　　处：《山东大学学报（医学版）》2007年第5期478-481,共4页Journal of Shandong University:Health Sciences

基　　金：山东省自然科学基金资助项目(Y2004C14)

摘　　要：目的:观察月龄、来源部位对犬骨髓MSCs体外增殖能力及成软骨细胞分化能力的影响,对软骨组织工程种子细胞的选择提供参考。方法:体外分离、培养4月龄、12月龄Beagle犬髂骨和胫骨骨髓中的MSCs,向软骨细胞诱导分化。依靠形态学观察、原代集落生成率、MTT法测生长曲线等方法观察各组MSCs的增殖能力,通过GAG含量、Ⅱ型胶原免疫组化比较诱导成软骨细胞效果。结果:4月龄犬髂骨与胫骨来源MSCs体外增殖特性及诱导成软骨细胞特性比较差异无统计学意义;12月龄犬髂骨组MSCs较胫骨组原代融合时间短、集落生成率高、生长曲线左移,但成软骨细胞特性无差异。相同部位比较,4月龄来源MSCs均较12月龄表现出较高的增殖能力和成软骨细胞分化能力,差异具有统计学意义。结论:成年犬不同部位来源MSCs增殖能力不同,分化能力相同;幼龄犬MSCs增殖、分化能力均高于成年犬。Objective： To explore the effect of age and origin on the proliferation and the chondrogenic differentiation capability of canine mesenchymal stem cells（MSCs） in vitro. Methods： MSCs isolated from bone marrows of the iliac bone and the shin bone in 4-month and 12-month canines, were cultured and induced into chondrocytes in vitro. The growth, purification, proliferation and chondrogenic differentiation abilities of MSCs in 4 groups were determined by using the clone forming rate and the MTF method. The GAG content and the positive cell rate of collagen Ⅱ were determined. Results： The proliferation and the chondrogenic differentiation abilities between MSCs isolated from iliac bones and shin bones were not statistically different in 4-month canines. In the 12-month canine groups, MSCs showed a shorter primary culture time and a higher clone-forming unit rate from iliac bones than from shin bones, whereas the expression of GAG and collagen Ⅱ was not positively different. Both the proliferation and the chondrogenic differentiation abilities were higher in the 4-month groups than in the 12-month groups. Conclusion： The proliferation ability of MSCs from both sites is different in the 12-month canines, while the chondrogenic differentiation capability is not significantly different. The proliferation and the chondrogenic differentiation capabilities of the 4-month canines are higher than those of the 12-month canines.

关 键 词：间充质干细胞 犬 年龄 增殖 成软骨分化 

分 类 号：R318[医药卫生—生物医学工程]