摘要
目的了解构建组织工程化骨的人间充质干细胞(MSCs),在体外成骨诱导环境中培养2代后,成骨细胞、软骨细胞、脂肪细胞的比例。方法体外扩增培养人MSCs,然后在含地塞米松、β-甘油磷酸钠、维生素C等的培养液中诱导培养2代,进行成骨细胞、软骨细胞、脂肪细胞的鉴定。结果人MSCs体外成骨诱导培养2代后,碱性磷酸酶阳性细胞数65%,Ⅰ型胶原免疫组化阳性细胞数55%、骨钙素阳性细胞数52%,Ⅱ型胶原免疫组化染色为阴性,油红O法染色阳性细胞数2.5%。结论人MSCs体外成骨诱导培养2代后,可有约50%的成骨细胞,少量脂肪细胞形成,无成熟的软骨细胞出现。该结论可为利用间充质干细胞构建组织工程化骨提供理论参考。
Objective To evaluate the proportion of osteob lasts,chondrocytes and adipocytes after human mesenchymal stem cells(MSCs )had been cultured under condition of osteogenic induction with chemicals.Methods Culture-expanded MSCs,stimulated with any chemicals(Dexamethasone,β-Glycerphosphoric sodium,Vitamin C)during the two passages and examined with histology for alkaline phos-phatase or lipid,stained immunohis tochemically for collagenⅠ,collagenⅡand osteocalcin.Results The percentage of cells expressed al-kaline phosphatase was 65%,expressed collagenⅠwas 55%,expressed osteocalcin was 52%,existed big lipid was 2.5%and non e of cells expressed collagenⅡ.Conclusion Culture-expanded MSCs and differen tiated into osteoblasts,there were half of cells expressed phenotype of osteoblast,a few cells expressed phenotype of adipocyte and no cells e xpressed phenotype of chondrocyte.
出处
《中国临床康复》
CSCD
2002年第22期3337-3338,I001,共3页
Chinese Journal of Clinical Rehabilitation
基金
全军十五医药卫生科研重点基金资助项目(01Z072)
关键词
间充质干细胞
细胞培养
成骨细胞
组织工程
mesenchymal stem cells
cell cultur e
osteoblasts
tissue engineering
