CAJ | 학술논문

背景:近年研究者发现胰岛素样生长因子1还可以诱导骨髓来源间充质干细胞向软骨细胞方向分化,但目前尚未见胰岛素样生长因子1诱导脂肪来源间充质干细胞向软骨细胞方向分化及在此过程中与转化生长因子β1相瓦作用的报道.目的:观察胰岛素样生长因子1诱导脂肪来源间充质干细胞向软骨细胞定向分化的可能性及在诱导分化中和转化生长因子β1的相互作用.方法:获取脂肪来源间充质干细胞,以2×10~5 cells/cm~2的密度接种于培养瓶,使用含胰岛素样生长因子1或(和)转化生长因子β1的无胰岛素软骨诱导剂诱导脂肪来源间充质干细胞.2周后获取细胞,制备细胞爬片,进行甲苯胺蓝染色和Ⅱ型胶原免疫组织化学染色,观察细胞内高硫酸化的蛋白聚糖和Ⅱ型胶原着色情况.RT-PCR检测Ⅱ型胶原蛋白、aggrecan及Sox9mRNA的表达.结果与结论:加入诱导剂后,甲苯胺蓝染色显示3个诱导组细胞呈多角形,胞浆及胞膜呈蓝色异染.Ⅱ型胶原免疫组织化学染色显示3个诱导组细胞胞浆及胞膜呈棕黄色着色.RT-PCR检测显示胰岛索样生长因子+转化生长因子组Ⅱ型胶原蛋白、aggrecan、Sox9 mRNA的表达均显著强于胰岛素样生长因子组和转化生长因子组,胰岛素样生长因子组和转化生长因子组显著强于对照组,而胰岛素样生长因子组与转化生长因子组差异无显著性意义.提示胰岛素样生长因子1可以单独诱导脂肪来源间充质干细胞向软骨细胞分化,表达软骨细胞特异性细胞表型,胰岛素样生长因子1与转化生长因子β1诱导脂肪来源间充质干细胞向软骨细胞分化有协同作用.
배경:근년연구자발현이도소양생장인자1환가이유도골수래원간충질간세포향연골세포방향분화,단목전상미견이도소양생장인자1유도지방래원간충질간세포향연골세포방향분화급재차과정중여전화생장인자β1상와작용적보도.목적:관찰이도소양생장인자1유도지방래원간충질간세포향연골세포정향분화적가능성급재유도분화중화전화생장인자β1적상호작용.방법:획취지방래원간충질간세포,이2×10~5 cells/cm~2적밀도접충우배양병,사용함이도소양생장인자1혹(화)전화생장인자β1적무이도소연골유도제유도지방래원간충질간세포.2주후획취세포,제비세포파편,진행갑분알람염색화Ⅱ형효원면역조직화학염색,관찰세포내고류산화적단백취당화Ⅱ형효원착색정황.RT-PCR검측Ⅱ형효원단백、aggrecan급Sox9mRNA적표체.결과여결론:가입유도제후,갑분알람염색현시3개유도조세포정다각형,포장급포막정람색이염.Ⅱ형효원면역조직화학염색현시3개유도조세포포장급포막정종황색착색.RT-PCR검측현시이도색양생장인자+전화생장인자조Ⅱ형효원단백、aggrecan、Sox9 mRNA적표체균현저강우이도소양생장인자조화전화생장인자조,이도소양생장인자조화전화생장인자조현저강우대조조,이이도소양생장인자조여전화생장인자조차이무현저성의의.제시이도소양생장인자1가이단독유도지방래원간충질간세포향연골세포분화,표체연골세포특이성세포표형,이도소양생장인자1여전화생장인자β1유도지방래원간충질간세포향연골세포분화유협동작용.
BACKGROUND:Recently,researches have found that insulin-like growth factor-1(IGF-1)can induce the differentiation of bone marrow-derived mesenchymal stem cells(BMSCs)into chondrocytes,but there are no reports concerning the differentiation of adipose-derived mesenchymal stem cells(ADMSCs)into chondrocytes induced by IGF-1,as well as interaction with transforming growth factor-β1(TGF-β1)during this process.OBJECTIVE:To explore the possibility of inducing ADMSCs chondrogenic differentiation by using IGF-1 and the interaction with TGF-β1 in induction.METHODS:ADMSCs were obtained,and seeded at 2×10~5 cells/cm~2 in culture flask.Insulin-free chondrogenic media containing IGF-1 or(and)TGF-β1 were used to induce ADMSCs.2 weeks later,cells were harvested and stained by using toluidine blue and collagen Ⅱ antibody immunohistochemistry.Intracellular sulfated proteoglycan and collagen Ⅱ coloring were observed.Reverse transcription-polymerase chain reaction(RT-PCR)was used to detect the expression of collagen Ⅱ,aggrecan and Sox9 mRNA.RESULTS AND CONCLUSION:After induced,toluidine blue stain exhibited that the cells in the three induction groups were polygonal,with cytoplasm and cell membrane of blue different dyeing.Immunohistochemistry for type Ⅱ collagen demonstrated that cytoplasm and cell membrane were stained brown in three induction groups.RT-PCR revealed that the expression of collagen Ⅱ,aggrecan,Sox9 mRNA of IGF + TGF group were significantly greater than the IGF and TGF groups,and IGF and TGF groups were significantly stronger than the control group.No significant difference was determined between the IGF and TGF groups.These results indicated that IGF-1 can induce chondrogenic differentiation from ADMSCs,expressing chondrocyte specific cell phenotype.There is synergism of IGF-1 and TGF-01 to induce the differentiation of ADMSCs into chondrocytes.