中国组织工程研究Issue(41)：6566-6571,6.DOI:10.3969/j.issn.2095-4344.2015.41.002
人髌下脂肪垫来源脂肪间充质干细胞的分离、培养及鉴定
Human adipose-derived mesenchymal stem cells from the infrapatellar fat pad：isolation, culture and identification
刘玉平 1刘涛 1王明明 1李明 1俞光荣2
作者信息
- 1. 滕州市中心人民医院创伤骨科，山东省滕州市 277500
- 2. 同济大学附属同济医院足踝外科，上海市 200065
- 折叠
摘要
Abstract
BACKGROUND:Infrapatelar fat pad is often partialy resected in the knee surgery, which can be used as an important source of adipose-derived mesenchymal stem cels. OBJECTIVE: To explore the strategies of isolation, culture, and identification of adipose-derived mesenchymal stem cels from the infrapatelar fat pad and to detect the expression of cel surface markers of human adipose-derived stem cels. METHODS: Infrapatelar fat pad was obtained from patients undergoing knee arthroscopy surgery, and attached cels were obtained from adipose tissue by using colagenase I. Cels were cultured in 10% low-sugar DMEM. Stem cels proliferation was detected by means of MTT and then, cel growth curve was made. The obtained cels were induced and differentiated into adipocytes and osteocytes. Expressions of cel surface markers CD29 and CD44 were detected. RESULTS AND CONCLUSION:A few of attached cels were observed after cultured 24 hours. Cels proliferated faster and exhibited spindle shape after 1 week. Cel adherence and proliferation were speeded up after subculture. Growth curve of cels exhibited that the passages 5 and 2 cels had higher reproductive activity than passage 8 cels. The obtained cels can be induced and differentiated into adipocytes and osteocytes. Results from flow cytometry showed that 96.8% passage 5 cels expressed CD29 and 97.6% expressed CD44. These findings indicate that high-purity adipose-derived mesenchymal stem cels with high reproductive ability are easy to be isolated from the infrapatelar fat pad, which may be a kind of ideal seed cels for cartilage tissue engineering.关键词
干细胞/脂肪干细胞/脂肪间充质干细胞/细胞培养/种子细胞/表面抗原Key words
Stem Cels/Mesenchymal Stem Cels/Cel Culture Techniques/Antigens, Surface/Tissue Engineering分类
医药卫生引用本文复制引用
刘玉平,刘涛,王明明,李明,俞光荣..人髌下脂肪垫来源脂肪间充质干细胞的分离、培养及鉴定[J].中国组织工程研究,2015,(41):6566-6571,6.
