解放军医学杂志2012，Vol.37Issue(6)：602-605,4.
悬滴培养对人脂肪间充质干细胞成骨分化的影响
Hanging-drop culture promote osteogenic differentiation of human adiposetissue-derived mesenchymal stem cells
杨静 1张琪2
作者信息
- 1. 121001辽宁锦州 辽宁医学院研究生院
- 2. 100039北京 北京武警总医院口腔颌而外科
- 折叠
摘要
Abstract
Objective To investigate the effect of hanging-drop culture on osteogenic differentiation of human adipose-derived mesenchymal stem cells (hADMSCs). Methods hADMSCs were isolated by enzyme digestion and cultured. The primary culture and second passage of hADMSCs were observed with phase-contrast microscope. To confirm the immunophenotype of hADMSCs, surface markers of CD44, CD90, CD 105, CD34 and CD4S were detected by flow cytometry. hADMSCs were then cultured in the hanging drops. The phenotypes of hADMSCs spheroid aggregate in hanging drops were then observed with phase-contrast microscope. After the adherence of cells, hADMSCs were cultured in osteogenic differentiation medium to achieve osteogenic differentiation. The expression of Runx2 and Osteopontin was detected by reverse transcription-polymerase chain reaction (RT-PCR) at 7th and 21st days. Results Most of hADMSCs grew as spindle-shaped cells with long cytoplasmic processes as observed under the phase-contrast microscope. hADMSCs shared the immunophenotypes of both multipotential stem cell and embryonic stem cell markers, as they were positive for CD44, CD90 and CD 105, and negative for CD34 and CD45 expressions by flow cytometry analysis, hADMSCs cultured in the hanging drops could form cell aggregates after 24 hours of culture, and could form three-dimensional spheroid aggregates with uniform size after 72 hours of culture as observed under the phase-contrast microscope. RT-PCR showed that the expression levels of Runx2 and Osteopontin in hanging-drop cultures were much higher than that of control. Hanging-drop culture could promote osteogenic differentiation of hADMSCs. Conclusions Hanging-drop culture is beneficial in promoting the efficiency of osteogenic differentiation of human adipose-derived mesenchymal stem cells.关键词
间质干细胞/细胞培养技术/细胞分化/骨生成Key words
mesenchymal stem cells/ cell culture techniques/ cell differentiation/ osteogenesis分类
医药卫生引用本文复制引用
杨静,张琪..悬滴培养对人脂肪间充质干细胞成骨分化的影响[J].解放军医学杂志,2012,37(6):602-605,4.
