摘要
目的比较牛去蛋白脱钙皮质骨基质（bDpDBM）微颗粒和牛脱钙皮质骨基质（bDBM）微颗粒用于体外干细胞培养的优缺点，探讨其作为微载体制备候选材料的可行性。方法冷冻粉碎牛四肢皮质骨，筛选出直径为200～500μm的微颗粒，按照常规方法进行脱脂、脱钙、脱蛋白处理。设bDpDBM和bDBM微颗粒为研究对象，用于静态条件下骨髓基质干细胞（BMSCs）的培养，比较两组微颗粒对于细胞增殖及功能保持的影响，分析其用于BMSCs培养的可行性。结果两组微载体进行培养第14天后显示bDpDBM组细胞扩增较bDBM组快，差异有统计学意义（P〈0．05）。经bDBM组培养的BMSCs的ALP定量及胞内钙含量明显较bDpDBM组多，差异有统计学意义（P〈0．05）。而bDpDBM颗粒培养的BMSCs经诱导可向成骨、成脂及成软骨三系分化。结论bDpDBM由于去除了脱钙骨基质中的活性诱导成分，培养增殖效率增高，可较好地保持干细胞表型，适宜用作体外大量扩增BMSCs的微载体候选材料。
Objective To discuss the potential of granular bovine deproteinized decalcified cortical bone matrix （bDpDBM） as a candidate for microcarriers by comparing bDpDBM and bovine decalcified cortical bone matrix （bDBM） in culture of bone marrow stromal stem cells （BMSCs） in vitro. Methods The cortical bone of bovine extremities was frozen and ground up to screen out granules of 200 - 500 μm in diameter. The tiny granules were degreased, decalcified and deproteinized in a conventional manner, bDpDBM and bDBM granules were used as scaffolds for static culture of BMSCs. The two groups were compared in terms of proliferation and maintenance of the multi-potentials of BMSCs. Results After 14 days＇ culture the proliferation of BMSCs in the bDpDBM group was significantly faster then in the bDBM group （ P 〈 O. 05）. The ALP expression and calcium deposition of BMSCs in the bDBM were significantly higher than in the bDpDBM group （ P 〈 0. 05） . The BMSCs cultured in the bDpDBM group could be induced into osteogenie, adipogenic and chondrogenic differentiations, indicating high maintenance of multi-potentials of BMSCs. Conclusion Because bDpDBM granules may facilitate highly efficient culture of BMSCs and can preferably maintain the multi-potentials of BMSCs due to removal of components with inductive activity, they may be used as a good candidate microcarrier in cell culture in vitro.
出处
《中华创伤骨科杂志》
CAS
CSCD
北大核心
2013年第4期336-340,共5页
Chinese Journal of Orthopaedic Trauma
基金
国家重点基础研究发展计划（973计划）（2009CB930003）
关键词
骨基质
骨髓细胞
组织支架
去蛋白
Bone matrix
Bone marrow cells
Tissue scaffolds
Deproteinization
