海藻酸钠-壳聚糖微囊包封小鼠骨髓间充质干细胞的体外研究

An in vitro study of mouse bone marrow mesenchymal stem cells encapsulated in alginate-chitosan-alginate microcapsules

目的探究海藻酸钠-壳聚糖微胶囊(alginate-chiotsan-alginate,ACA)包封小鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)后细胞生长及增殖能力。方法C57小鼠采用全骨髓法进行骨髓间充质干细胞的原代提取。配制1.5%和0.05%海藻酸钠溶液、1.1%氯化钙溶液、0.6%壳聚糖溶液(pH 6.3)和55 mmol/L柠檬酸钠溶液(pH 5.6)用高压静电微囊法制取包封小鼠骨髓间充质干细胞的海藻酸钠-壳聚糖微胶囊,并对微囊进行强度及通透性检测。通过Calcein-AM/PI进行ACA包封BMSCs的活死细胞染色,CCK8检测ACA包封BMSCs的细胞增殖能力。结果获得均匀一致,直径约300μm的包封BMSCs的海藻酸钠-壳聚糖微胶囊,Calcein-AM/PI染色及CCK8检测发现BMSCs可在ACA中存活及稳定增殖。结论海藻酸钠-壳聚糖微胶囊具有良好的生物相容性,可使BMSCs在微胶囊中生长及增殖,为细胞微胶囊应用于各生物医学领域提供便利。

Objective To investigate the proliferation of bone marrow mesenchymal stem cells(BMSCs)encapsulated in alginate-chitosan-alginate(ACA)microcapsules.Methods C57 mice were used to culture bone marrow mesenchymal stem cells(BMSCs)using whole bone marrow method.1.5%sodium alginate solution,0.05%sodium alginate solution,1.1%calcium chloride solution,0.6%chitosan solution(pH 6.3)and 55 mmol/L sodium citrate solution(pH 5.6)were prepared and the bone marrow mesenchymal stem cells(BMSCs)encapsulated in alginate-chitosan-alginate(ACA)microcapsules.The strength and permeability of microcapsules were tested.Calcein-AM/PI was stained with live/dead cells,and CCK8 was used to detect cell proliferation of ACA encapsulated BMSCs.Results Alginate-chitosan-alginate microcapsules with uniform diameter of about 300μm were obtained.Calcein-AM/PI staining and CCK8 assay showed that BMSCs could survive and proliferate steadily in ACA microcapsules.Conclusion Alginate-chitosan-alginate microcapsules have good biocompatibility,which can make BMSCs proliferate in microcapsules and provide convenience for cell microcapsules in various biomedical fields.