自体骨髓间质干细胞外基质支架在体外软骨组织工程中的应用  

Application of autologous bone marrow mesenchymal stem cell-derived extracellular matrix scaffold on tissue engineering cartilage in vitro

作　　者：唐成[1] 徐燕[1] 王黎明[1] 苗登顺[2] 裴璇[3] 魏波[1] 杜小涛[1] 金成哲[1] 

机构地区：[1]南京医科大学附属南京医院骨科,南京210006 [2]南京医科大学基础医学院,南京210006 [3]东南大学生物科学与医学工程学院

出　　处：《中华骨科杂志》2013年第6期649-656,共8页Chinese Journal of Orthopaedics

基　　金：国家自然科学基金（31070861,81171745）

摘　　要：目的探讨利用自体骨髓间质干细胞外基质（autologous bone marrow mesenchymal stemcell-derived extracellular matrix，aBMSC-dECM）支架体外制备组织工程软骨的可行性。方法取2周龄新西兰大白兔5只，分离、培养骨髓间质干细胞，原代培养4周，收集其分泌的细胞外基质，制备aBMSC-dECM支架。对支架行扫描电镜和HE染色观察。分离培养自体软骨细胞，植人支架内，48h后对细胞一支架复合物行Live-Dead染色。分别于种植后1、2、4和6周对细胞一支架复合物（组织工程软骨）进行大体观察、体积测量、HE染色、Safranin-0染色、Ⅱ型胶原免疫组织化学染色、Real-TimePCR检测和抗压强度测试。Objective To evaluate the feasibility of the formation of tissue engineering cartilage in vitro by using autologous bone marrow mesenehyma] stem cell-derived extracellular matrix （aBMSC-dECM） scaffold. Methods The autologous bone marrow mesenchymal stem cells were harvested from 5 New Zealand white rabbits which are two weeks old. The extracellular matrix was collected after 4 weeks culture and fabricated into aBMSC-dECM scaffold. The scaffold was investigated by a scanning electron microscope and HE staining. The autologous articular chondrocytes were isolated, cultured and seeded into the aBMSC- dECM scaffold. Live-Dead staining was analyzed 48 h after seeding. Gross morphological and volume mea- surement, HE staining, Safranin-O staining and type Ⅱ collagen immunohistochemistry staining, RT-PCR assay, and compression strength test were operated for the cell-scaffold composite at 1, 2, 4, and 6 weeks af- ter the cultivation respectively. Atelocollagen scaffold was used as the control group.

关 键 词：间质干细胞 软骨 组织工程 体外研究 

分 类 号：R318.08[医药卫生—生物医学工程]