摘要
目的 观察人骨髓间充质干细胞的生长特点及诱导条件下的成骨能力。方法 抽取人骨髓组织 ,梯度离心后 ,保留贴壁细胞传代 ,稳定传代后改用含地塞米松、β 甘油磷酸钠和维生素C的诱导培养液 ,通过倒置显微镜观察、碱性磷酸酶染色、骨钙素免疫组化染色等方法进行观测。结果 原代人骨髓间充质干细胞 10～ 12d左右即可长满 ,并可稳定传代 ,传代细胞 5～ 6d即可传代 ,在诱导培养液中 2周形成多层结构 ,并聚集成细胞钙化结节。诱导培养细胞碱性磷酸酶染色 ,骨钙素免疫组化染色阳性。结论 人骨髓间充质干细胞经合理的体外诱导培养后 ,符合成骨细胞的形态特征和生物学特性 ,可为体外构建组织工程化骨提供自体种子细胞。
Objective To observe the growth and osteogenic property of cultured human mesenchymal stem cells(hMSCs ).Methods The bone marrow was obtained by aspiration from a healthy male volunteer aged 26,mononuclear cells were then isolated by centrifugation in Percoll density gradient.The adhesive cells were preserved. hMSCs were cultured in a condition medium in subculture including dexamethasone,vitamin C,and β-glycerophosphate,and cytochemical staining for alkaline phosphatase(ALP),cytochemical staining for osteocalcin expression and invert microscope observing were applied to measure the hMSCs proliferation capability and osteogenic property.Results MSCs reached confluence and started to form multilayers on the 10th or 12th day,passaged stably as well.The hMSCs passaged in 5 or 6 days,and transformed into multilayer Structure in 2 weeks and gradually congregated into a mineralized matrix.The hMSCs in induced culture showed cytochemical staining for ALP and osteocalcin expression.Conclusion One of important biological effects of the induced hMSCs is accelerate hMSCs differentiate into osteoblast, which provides autogenous seeding cells for in vitro construction of tissue engineered bone.
出处
《江西医学院学报》
2004年第2期1-3,F002,共4页
Acta Academiae Medicinae Jiangxi
基金
国家 8 6 3计划重大专项 (编号 2 0 0 3AA2 0 5 0 0 2 )
重庆市重点科技攻关基金资助项目 (编号 2 0 0 3 13 6 )
第三军医大学校科研基金资助项目 (编号XG2 0 0 32 7)
2 0 0 2年西南医院科技创新基金资助项目
关键词
骨髓间充质干细胞
地塞米松
成骨诱导
成骨分化
mesenchymal stem cells
dexamethasone
osteogenic induction
osteogenic differentiation
