摘要
骨髓间质干细胞 (mesenchymalstemcell,MSCs)是一类存在于骨髓中的具有多向分化潜能的干细胞 ,在体外不仅可分化为间质类细胞 ,而且可以分化为非间质细胞。本研究探讨人骨髓间质干细胞的体外分离纯化、培养扩增和向成骨细胞诱导分化的条件。利用密度为 1.0 73g/ml的Percoll分离骨髓的单个核细胞 ,以含 10 %胎牛血清的低糖DMEM培养基培养与扩增MSCs ,细胞纯度可达 95 %左右。取第二代和第三代的MSCs ,以含有不同浓度的抗坏血酸、β 磷酸甘油、地塞米松的诱导培养基向成骨细胞诱导 ,诱导后的细胞呈现典型的成骨细胞样改变 ,免疫组化技术显示其I型胶原染色阳性 ,ALP染色阳性 ,表明MSCs在体外具有向成骨细胞分化的潜力。
Human bone marrow derived mesenchymal stem cells(MSCs) are a population of cells capable of differentiating along multiple cell lineages. This study investigated the culture and in vitro expansion of human MSCs as well as requirements for osteogenic differentiation of MSCs. The mononuclear cells in each bone mar row samples were obtained by gradient centrifugation with Percoll at a density of 1.073g/ml and then cultured and expanded with low glucose Dulbecco′s modified Eagle′s medium(L DMEM) containing 10% fetal bovine serum(FBS). Flow cytometry analysis demonstrated a purity of MSCs about 95%. MSCs derived from the second and the third passages were cultured for 16 days in the media containing dexamethasone(Dex), ascorbic acid(ASA) and β glycerophosphate(β GP) of various combinations. The induced MSCs changed their morphology to that of osteoblast type. Immunohistochemical studies proved strong college I expression and ALP stain was positive, suggesting that the MSCs were induced to differentiate along osteoblasts lineage.
出处
《中国生物医学工程学报》
CAS
CSCD
北大核心
2002年第3期251-255,共5页
Chinese Journal of Biomedical Engineering
基金
国家航天 863领域项目 (863 2 7 2 15 )
国家自然科学基金青年基金资助项目 (3 990 0 0 3 6)
关键词
间质干细胞
骨髓
成骨细胞
分化
实验研究
Mesenchymal stem cells
Bone marrow
Osteoblasts
Differentiation
