骨髓间充质细胞与速固化磷酸钙支架共培养的实验研究

[目的]观察人骨髓间充质干细胞在速固化磷酸钙骨水泥支架中的增殖和生长情况。[方法]将速固化壳聚糖-磷酸钙骨水泥支架材料预先成形。采用全骨髓法分离人骨髓间充质细胞并在体外对细胞进行培养和扩增。72h首次换液,细胞融合80%传代,取第3代细胞用于实验。将细胞与磷酸钙骨水泥支架复合培养,分别利用四甲基偶氮唑蓝MTT法及扫描电镜等检测细胞在材料中的增殖和生长情况。[结果]用酶标仪检测OD值提示共培养后,支架内的细胞数量逐渐增加。扫描电镜结果显示支架材料内部为多孔结构,孔内见有细胞生长,细胞表面可见分泌颗粒。[结论]速固化壳聚糖-磷酸钙骨水泥对人骨髓间充质干细胞有较好的细胞相容性,细胞可在材料内黏附和增殖,是理想的骨组织工程支架材料。

Experimental study on union culture of mesenchymal stem cell with fast setting calcium phosphate scaffold

[Objective]To examine the effects of fast setting calcium phosphate scaffold on human mesenchymal stem cells(MSCs) adhesion and proliferation.[Method]Scaffolds were self-hardened in molds.Three ml human bone marrow aspirates,taken form the iliac crest of normal donors were suspended in low glucose DMEM supplemented with 10% fetal bovine serum.Atfer 72 hours the nonadherent cells were removed by changing medium,and the adherent layer cultured until it reached 80% conference.Cultured cells at passage 3 were used for the experiments.MSCs were seeded on calcium phosphate scaffold.The cell-scaffold constructs were cultured with complete media for 1 week.Scanning electron microscope(SEM)and the terazolium-based colorimetric assay(MTT test)examinations were performed.[Result]When cells were cultured in scaffolds in vitro,the number of cells on scaffold increased with time.SEM showed that the scaffold had a three-dimensional interconnected pore structure.The cells appeared to infiltrate into the macropores of the scaffold and the extracellular matrix components were observed.[Conclusion]This study demonstrates that the fast setting calcium phosphate scaffold has good biocompatibility,cells are able to adhere and proliferate on scaffold specimens,and it is a good substrate candidate for tissue engineered bone substitute.