骨髓间充质干细胞与β-磷酸三钙多孔陶瓷体外复合培养实验研究  

In vitro co-culture of bone marrow mesenchymal stem cells and β-porous calcium phosphate ceramics

作　　者：曾宪海[1] 李娟娟[1] 赵海亮[1] 

机构地区：[1]深圳市龙岗中心医院,广东深圳518172

出　　处：《临床医药实践》2013年第8期607-610,共4页Proceeding of Clinical Medicine

摘　　要：目的:研究犬骨髓间质干细胞(BMSCs)与β-磷酸三钙多孔陶瓷(β-TCP)的体外复合培养。方法:体外培养BMSCs作为种子细胞,β-TCP作为组织工程支架,制备β-TCP-BMSCs复合物,倒置显微镜下观察骨髓间充质干细胞培养传代过程中细胞形态变化及细胞在材料孔隙区的贴附及生长情况,流式细胞仪鉴定骨髓间充质干细胞表面抗原CD44、CD45表达情况。结果:相差倒置显微镜下观察:原代、第2代、第3代及第4代培养的细胞形态,贴壁细胞逐渐由成纤维细胞样向多形化方向转化(三角形、菱形等),最终重叠生长形成结节状;将细胞接种于材料上进行培养,见孔隙间有大量交叠细胞生长增殖。扫描电镜下观察:在材料表面细胞呈梭形,并融合成片状,表面见丝状突起,在孔隙边缘的细胞有大量胶原和细胞外基质分泌。流式细胞仪测定骨髓间充质干细胞表面抗原CD44阳性表达率(89.12±1.57)%,CD45阳性表达率为(2.32±0.45)%。结论:β-TCP与细胞有较好的亲和力,能够为基质细胞增殖、分化和成骨提供自然的三维空间,可作为组织工程的支架材料应用于骨和软骨缺损的修复。Objective：To study the biocompatibility between the bone marrow mesenchymal stem cells（BMSCs） of beagle with p-porous calcium phosphate ceramics （β-TCP） . Methods：Through vitro inducement cultivation, the BMSCs of Beagles was used as seed cell. β-TCP as the tissue engineering bracket ,the β-TCP with BMSCs complexion was made. To observe the cell storage and growing condition on material pore with scanning electric microscope and phase contract inversion electronic micro- scope. The positive surface marker CD44 and negative marker CD45 of ceils were detected for the identification of BMSCs with flow cytometry. Results：According to observation from the phase contrast inversion electric microscope primary cultivate from passage I to passage IV, adherent depend-cell convert into fibroblast-like. Triangle diamond and polygon ultimately formation nodosity through overlap growing cell will be into material. Cultivated for 4 days ceils generated dramatically. In the material pore showing the coincide growing observing through scanning electronic microscope finger-shaped cell from the material sur- face confluent lamella. The surface had filamentous protrusion. The bargain of pore had a plenty of collagen and extra-cellular matrix secret. The results of flow cytometry showed that CD44 expression of BMSCs was（ 89.12 ± 1.57 ） % , CD45 positive ex- pression rate was （2.32 ± 0.45 ） %. Conclusion ： β-TCP has good affinity with adherent depend-cell and can be providing natu- ral the 3-dimesional space for matrix cell generation ossification differentation.

关 键 词：组织化工程技术 骨髓间充质干细胞 β-磷酸三钙多孔陶瓷 

分 类 号：R318[医药卫生—生物医学工程]