几丁质支架对体外间质干细胞成软骨分化的影响  

Influence of chitosan scaffolds on chondrogennic differentiation of mesenchymal stem cells in vitro

作　　者：付勤[1] 王广斌[1] 邴佩旭[2] 李斌[3] 傅永慧[1] 

机构地区：[1]中国医科大学附属二院第一骨外科,沈阳110004 [2]沈阳市第五人民医院 [3]北京市海淀区医院

出　　处：《中国骨质疏松杂志》2007年第9期638-641,共4页Chinese Journal of Osteoporosis

基　　金：辽宁省自然科学基金资助项目(20042069)

摘　　要：目的鉴定间充质干细胞诱导分化为软骨细胞后,其在单层培养和几丁质支架上培养的差别。方法密度梯度离心兔股骨骨髓,分离BMSCs,用TGF-β1诱导第3代的BMSCs向软骨方向分化,2周后检测Ⅱ型胶原表达情况。将分化后的软骨样细胞传代,用无TGF-β1的培养基分别进行单层培养和接种在几丁质支架上培养2周,再用免疫组化检测Ⅱ型胶原表达情况。结果BMSCs经TGF-β1诱导后向软骨细胞方向分化,诱导后的细胞在无TGF-β1的培养中,采用单层培养方式的软骨细胞很快出现去分化表型,而接种在几丁质支架上的细胞仍能较好表达Ⅱ型胶原。结论几丁质支架在延缓软骨细胞去分化和老化方面有积极作用。Objective This paper is intended to study the difference of the cartilage cells （which were induced by the mesenchymal stem cells first） cultured by means of monolayer culture and chitosan scaffolds respectively. Methods Bone marrow was obtained from the rabbit femora and BMSCs were separated from rabbit bone marrow by the method of density gradient centrifugation. TGF-β1 was used to induce chondrogenic differentiation of the third generation of BMSCs. Two weeks later, Collagen Ⅱ of the BMSCs was detected by immunohistochemistry, and then the cells induced by TGF-β1 were respectively cultured by means of monolayer culture and chitosan scaffolds for two weeks. The expression of Collagen Ⅱ was detected again by immunohistochemistry. Results After BMSCs were cultured with TGF-β1 , Collagen Ⅱ detected by immunohistochemistry proved that these cells showed the traits of cartilage cells. But when these cells continued to be cultured without TGF-β , they soon showed dedifferentiation of cartilage cells by the means of monolayer culture, whereas the cells in the chitosan scaffolds expressed Collagen Ⅱ normally. Conclusion After BMSCs were induced into cartilage cells, chitosan scaffolds play an important role in delaying aging and dedifferentiation of cartilage cells.

关 键 词：间充质干细胞 Ⅱ型胶原 软骨细胞 几丁质 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]