摘要
目的探讨体外培养条件下人脂肪干细胞增殖和分化过程中端粒酶的表达水平，为其作为种子细胞的应用研究提供理论基础。方法体外分离、培养人脂肪干细胞并传代，行流式细胞表面抗原分析，并用油红0染色及茜素红染色行成骨和成脂诱导分化的鉴定；采用TRAP法分别检测新鲜的不同时间培养的人脂肪干细胞和诱导分化为脂肪细胞的人脂肪干细胞的端粒酶活性。结果脂肪干细胞具有向脂肪细胞、成骨细胞多向分化的能力，且表达干细胞相关表面标志物。新鲜分离和传代的脂肪干细胞，在体外培养12代内端粒酶活性呈阴性或低水平表达；一旦经过成脂诱导分化，细胞端粒酶活性表达上调，培养3—6d后端粒酶活性开始出现逐渐降低。结论用胶原酶消化法从脂肪抽吸术中得到的细胞主要是人脂肪干细胞；在体外培养增殖的过程中，人脂肪干细胞的端粒酶活性未见异常表达；成脂诱导分化早期人脂肪干细胞端粒酶活性增高，其后开始出现下降趋势。
Objective To investigate the telomerase activity of human adipose derived stem cells （ADSCs） during proliferation and differentiation in vitro. Methods ADSCs were highly purified and cultured in vitro. The morphology, phenotype and biological properties of the cultured ADSCs were observed by flow cytometer. Then ADSCs were induced to differentiate into adipocytes and osteoblast. The telomerase activity was detected by TRAP. Results ADSCs had the ability of multi-directed differentiation, like adipocytes and osteoblast. It could also express the stem cell-related surface markers. The telomerase activity was negative or lowly expressed in ADSCs in Vitro within 12 generations. The telomerase activity was up-regulated when ADSCs was adipogenic differentiated, but deceased 3-6 days later. Conclusions The telomerase activity of ADSCs is not changed during culture in vitro. It is upregulated when ADSCs are induced to adipogenic differentiation, but decreased later.
出处
《中华整形外科杂志》
CAS
CSCD
北大核心
2010年第1期48-52,共5页
Chinese Journal of Plastic Surgery
关键词
脂肪细胞
间充质干细胞
端粒酶
细胞分化
Adipocytes
Mesenchymal stem cells
Telomerase
Cell differentiation
