摘要
联合应用胶原酶消化、贴壁筛选及单克隆培养法从人脂肪组织中分离、培养人脂肪干细胞(hADSCs),观察hADSCs的形态学特征。通过流式细胞术检测hADSCs表面抗原;通过细胞化学染色和免疫荧光观察hADSCs的一般形态学特征;利用透射电镜分析hADSCs的超微结构。结果显示体外分离、培养出高度同源性的hADSCs,hADSCs具有相对特异的细胞表面标志,即CD29、CD44、CD90、CD105及CD166呈阳性表达,而CD31、CD45及HLA-DR呈阴性表达;hADSCs苏木精-伊红染色见胞浆淡染,油红O及碱性磷酸酶(AKP)染色阴性;免疫荧光CD29、CD166呈阳性;透射电镜下可见hADSCs细胞质内粗面内质网、线粒体丰富,细胞表面可见丰富微绒毛。本实验结果为hADSCs在细胞、组织再生移植修复中的应用提供了形态学基础。
This paper is aimed to isolate and to cultivate human adipose-derived stem cells（hADSCs） from the adipose tissue by a combination of collagenase digestion,adherence to flasks and monoclonal cultural method so as to observe the morphological characteristics of the hADSCs.The immunophenotypes of hADSCs were detected by flow cytometry techniques. The general morphological characteristics of hADSCs were observed by cytochemical and immunofluorescent techniques.The ultrastructure of hADSCs was observed by transmission electron microscopy（TEM）.The experimental results showed that hADSCs had unique immunophenotypes and they were positive for CD29,CD44,CD90,CD105 and CD166,but negative for CD31,CD45 and HLA-DR.Cytochemistry showed that cytoplasm of hADSCs was stained with light blue by hematoxylin-eosin,negative for Oil red O and AKP,and positive for immunofluorescence CD29 and CD166.There were abundant organella and microvilli in the ultrastructure of hADSCs.The results validate that they will offer a morphological foundation for application of the hADSCs.
出处
《生物医学工程学杂志》
EI
CAS
CSCD
北大核心
2011年第2期337-341,共5页
Journal of Biomedical Engineering
基金
863重大专项基金资助项目(2004AA205020)
国家自然科学基金资助项目(30700872)
吉林大学青年教师基金资助项目(419070100050)
吉林省卫生厅资助项目
关键词
人脂肪干细胞
流式细胞术
激光扫描共聚焦显微镜
超微结构
Human adipose-derived stem cells（hADSCs）
Flow cytometry
Laser scanning confocal microscope
Ultrastructure
