摘要
目的:观察Sox9基因对人脂肪干细胞(ADSCs)的诱导作用。方法:分离、纯化、培养人源ADSCs,并绘制生长曲线,传代三次后的ADSCs利用脂质体转染Sox9基因,选用抗生素G418进行筛选。以空载体转染细胞作对照,分别取48h和14d转染后的细胞做Flag蛋白免疫组织化学鉴定。通过检测转染细胞中II型胶原来确定ADSCs是否向软骨细胞分化。结果:ADSCs呈长梭形,形态与骨髓间充质干细胞相似,600μg/mlG418为最适筛选浓度。转染后第48h和14d的细胞均能表达Sox9基因融合表达的Flag蛋白。第48h和14d,转染效率分别为93%和75%。转染后14d的ADSCs表达II型胶原,转染后48h实验组和对照组都为阴性。结论:Sox9基因能诱导脂肪干细胞向软骨细胞分化。
Objective： To investigate the inductive effect of Sox9 gene on chondrocyte differentiation of human adipose-derived stem cells. Methods： Human ADSCs were separated, purified and cultured in the incubator, and then the growth curve of ADSCs were described. Passage 3 of ADSCs was transfected with pcDNA3.1-Sox9 using Lipo-fectamineTM2000 and the transfected cells were selected by G418. After incubation for 48 h and 14 d of transfected ADSCs, the expressions of Sox9 were indirectly determined with Flag protein by immunochemical method. The chondrogenesis of ADSCs were determined by morphology and immunochemical detection of expression of type II collagen. Results： ADSCs were long spindle-shaped, and similar to bone marrow-derived mesenchymal stem cells. The exogenous expression of Sox9 was detected 48 h and 14 d after transfection and the rate of Sox9 positive ADSCs was about 93%and 75 %, respectively. The expression of type II collagen was found in the ADSCs after transfection for 14 d, but not in the control ADSCs and the ones after transfection for 48 h. Conclusion： Sox9 gene could induce chondrocyte differentiation of human adipose-derived stem cells.
出处
《现代生物医学进展》
CAS
2010年第20期3851-3853,共3页
Progress in Modern Biomedicine
关键词
SOX9基因
脂肪干细胞
软骨细胞分化
转染
原代培养
Sox9 gene； adipose-derived stem cells； chondrocyte differentiation； transfection； primary culture；
