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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
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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.
出处 《山东大学学报(医学版)》 CAS 北大核心 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)
关键词 间充质干细胞 年龄 增殖 成软骨分化 Mesenchymal stem cells(MSCs) Canine Age Proliferation Chondrogenic differentiation
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