摘要
通过共培养人脂肪干细胞(hADSCs)与脂肪细胞来诱导hADSCs成脂分化。将hADSCs和脂肪细胞分别采用1:5、1:1、2:1和5:1共4个比例,在Transwell中进行共培养。共培养8d后,用油红O和台盼蓝对共培养后的hADSCs进行染色,显微镜下观察细胞形态及其成脂分化情况。此外,将共培养时间延长至20d,进行油红O和台盼蓝染色、流式细胞仪分析及hoechst33342/PI死活染色以检测其成脂分化情况。实验结果表明:油红O和台盼蓝染色结果显示共培养8dhADSCs具有较高活性,但并不能成脂分化;而油红O和台盼蓝染色及hoechst33342/PI死活染色结果显示共培养20d后,hADSCs亦具有高活性但仍不能成脂分化;然而流式细胞仪分析结果表明,共培养20d后hADSCs的CD105表达有所减弱。研究结果表明:hADSCs与脂肪细胞共培养尚不能促使hADSCs成脂分化。
The differentiation of human adipose-derived stem cells（hADSCs） into adipocytes was studied by co-culturing them with human mature adipocytes.The transwell was utilized to indirect co-culture of hADSCs and human mature adipocytes,with 4 different kinds of hADSCs to mature adipocytes ratios i.e.1：5,1：1,2：1 and 5：1.After 8 days of co-culture,the Oil Red O and Trypan Blue staining were performed for the evaluation of adipogenic differentiation of hADSCs.In addition,the Oil Red O and Trypan Blue staining,flow cytometry analysis and hoechst33342/PI double staining were used after 20 days of co-culture.The Oil Red O and Trypan Blue staining showed that hADSCs with high viability could not differentiate into mature adipocytes after 8 or 20 days of co-culture.However,flow cytometric analysis indicated that CD105 expression of hADSCs decreased after 20 days of co-culture.These results indicated that hADSCs after co-culture could not successfully differentiate into adipocytes.
基金
国家自然科学基金资助项目(30670525/30700181)
高等学校博士学科点专项科研基金新教师基金资助项目(20070141055)
大连理工大学青年教师基金资助项目(893228)
关键词
脂肪细胞
脂肪干细胞
共培养
成脂分化
油红O染色
adipocytes
adipose-derived stem cells
co-culture
adipogenic differentiation
Oil Red O staining
