【摘要】 目的:探讨胰岛素样生长因子-I(IGF-I)和软骨细胞培养液体外诱导骨髓间充质干细胞(MSCs)向软骨细胞分化的可能性。方法:采用Percoll分离液分离培养人胚MSCs,体外扩增,用流式细胞仪测定MSCs的CD44、CD71、CD34、CD45表达;在第4代MSCs中加入的条件培养基进行诱导,根据条件培养基的不同分为4组:①IGF-I组:培养基中加入100ng/mLIGF-I;②软骨细胞培养液组:加入软骨细胞培养液;③联合诱导组:培养液中加入100ng/mLIGF-I和软骨细胞培养液;④对照组:培养基中不加入IGF-I和软骨细胞培养液。采用倒置显微镜观察、II型胶原免疫组化、细胞内蛋白多糖(PG)含量测定等方法判断诱导细胞的形态变化和表达软骨基质的能力。结果:体外培养的MSCs呈现成纤维细胞样形态,流式细胞仪检测结果显示,MSCs阳性表达CD44,阴性表达CD34,CD45;IGF-I和软骨细胞培养液联合诱导15d后,MSCs开始呈现软骨细胞的形态特点,II型胶原免疫组化呈阳性,而未诱导的MSCs组呈阴性;联合诱导组细胞内PG含量为8.92μg/ml,显著高于IGF-I组、软骨细胞培养液组和对照组,但低于正常软骨细胞(P<0.01)。结论:IGF-I和软骨细胞培养液能诱导MSCs向软骨细胞分化。更多还原

【Abstract】 Objective: To investigate the differentiation of human fetal mesenchymal stem cells (MSCs) exposed to insulin-like growth factors-I (IGF-I) and chondrocyte-derived medium into chondrocytes in vitro. Methods: Human fetal MSCs were isolated and cultured in DMEM. Cell surface antigen was analyzed by flow cytometry. And then MSCs were cultured with 100 ng/mL IGF-I medium(IGF-I group), chondrocyte-derived medium(chondrocyte-derived medium group), 100 ng/mL IGF-I plus chondrocyte-derived medium(combined treated group), or without IGF-I and chondrocyte-derived medium(control group)respectively. After 15 days, cultures were assessed by collagen types II immunohistochemistry and proteoglycan production. Results: The isolated and cultured human fetal MSCs displayed a fibroblast-like morphology. MSCs were uniformly positive for CD44, and did not express CD34 and CD45. MSCs in monolayer cultures treated with IGF-I and chondrocyte-derived medium exhibited a chondrocytic phenotype and expressed collagen types II. Proteoglycan contents in combined treated MSCs group were significantly higher than IGF-I group,chondrocyte-derived medium group and control group. However, proteoglycan production of MSCs treated with IGF-I and chondrocyte-derived medium remained lower than cultured chondrocytes. Conclusions: Human fetal MSCs can be induced and differentiated into chondrocyte-like cells when cultured in IGF-I and chondrocyte-derived medium.更多还原