西安交通大学学报（医学版）2012，Vol.33Issue(5)：587-592,6.
人骨髓间充质干细胞生物学特性及腺病毒介导的GFP标记
Biological features of human mesenchymal stem cells and labelling with Ad-GFP transfection
谭云鹤 1种铁 2杨平 1郭子宽 2甘为民 3陈长虹 1翟宇强 2石硕文 2彭秀君2
作者信息
- 1. 西安交通大学医学院第二附属医院泌尿外科,陕西西安710004
- 2. 西安市中心医院泌尿外科,陕西西安710003
- 3. 军事医学科学院放射与辐射医学研究所实验血液学研究室,北京 100850
- 折叠
摘要
Abstract
Objective To establish a method for isolating, culturing and identifying human mesenchymal stem cells (hMSCs) so as to investigate the transient expression by using Ad-GFP transfection. Methods hMSCs were isolated and purified from human bone marrow with Ficoll density gradient centrifugation and by adhering to the culture plastic. The proliferation ability was evaluated by MTT assay. ALP Kit and red oil O staining were used to prove the differentiation ability of osteoblasts and adipoblasts, respectively. The potential to differentiate into smooth muscle cells (SMCs) was assayed with α-SMA monoclonal antibody. Surface markers in hMSCs were detected by flow cytometry (FCM). Then Ad-GFP was transfected into hMSCs and the transient expression was examined with fluorescence microscope and FCM. Results hMSCs were fibroblast-like in morphology with a high proliferation, and homogenously expressed CD44, CD73, CD106 and PDGFRP were negative for CD14, CD31, and CD45. After induction, the cells were ALP and a-SMA positive and red oil O staining was positive, too. At 48-h culture, the transient expression was up to 99.81% when transfected at an MO! of 200. Conclusion Highly purified hMSCs can be obtained with Ficoll density gradient centrifugation. Ad-GFP is a preferable method to label cells and can be used as a source for tissue engineering in vivo.关键词
间充质干细胞/分化/细胞表面标志/Ad-GFP/细胞转染/人淋巴细胞分离液(Ficoll)Key words
mesenchymal stem cell/differentiation/cell surface marker/Ad-GFP/cell transfection/Ficoll density gradient centrifugation分类
医药卫生引用本文复制引用
谭云鹤,种铁,杨平,郭子宽,甘为民,陈长虹,翟宇强,石硕文,彭秀君..人骨髓间充质干细胞生物学特性及腺病毒介导的GFP标记[J].西安交通大学学报（医学版）,2012,33(5):587-592,6.
