兔脂肪干细胞在含有转化因子和转铁蛋白培养基中向软骨细胞的分化

Chondrogenic differentiation of rabbit adipose-derived stem cells in culture medium containing transforming factors and transferrin

背景:成软骨的种子细胞的选择是软骨组织工程研究中的关键因素。目的:观察脂肪干细胞在含有转化因子和转铁蛋白诱导条件下向软骨细胞分化的能力。方法:取新西兰大白兔颈背部脂肪组织,机械分离及酶消化法获得脂肪干细胞,显微镜下观察细胞黏附及生长情况;加入含有转化因子β1和转铁蛋白的诱导培养基培养2周后以免疫组化方法检测Ⅱ型胶原的表达。结果与结论:从兔脂肪组织中分离出的干细胞原代培养时24h贴壁,96h后达80%融合;于软骨诱导培养基内向软骨诱导7d后形成软骨结节,14d后Ⅱ型胶原免疫组化阳性。结果初步表明兔脂肪干细胞经诱导后可以向软骨细胞分化。

BACKGROUND：The choice of seed cells is a key factor to the cartilage tissue engineering.OBJECTIVE：To explore the chondrogenic potential of adipose-derived stem cells（ADSCs） in culture medium containing transforming factors and transferring.METHODS：ADSCs were obtained from the subcutaneous adipose tissue of 6-month-old New Zealand white rabbits＇ neck back by mechanical digestion and enzyme digestion,and then cultured and amplified in vitro.The adhesion and growth of cells were observed using inverted phase contrast microscope.Two weeks after the cells were cultured in chondrogenic medium,collagen type Ⅱ expression was detected by immunohistochemistry staining.RESULTS AND CONCLUSION：The primary stem cells isolated from adipose tissue adhered on the plate in 24 hours,and reached 90% confluence in single layers after 96 hours.The cartilage nodules were observed at 7 days after ADSCs were cultured under chondrogenic medium,while collagen type Ⅱ was positive 14 days later.ADSCs of rabbits can express chondrogenic phenotype when they were cultured in chondrogenic medium,which proves the ADSCs can be one of the options for cartilage tissue engineering.