胰岛素样生长因子诱导外胚间充质细胞向成骨细胞分化的实验研究

发布时间：2018-07-05 18:09

本文选题：外胚间充质细胞 + 成骨细胞　；参考：《昆明医学院》2008年硕士论文

【摘要】： 目的:建立SD胎鼠颌突未分化外胚间充质细胞的体外培养细胞模型,在此基础上探讨胰岛素样生长因子-1(IGF-1)诱导胎鼠颌突外胚间充质细胞向成骨细胞(OB)分化的作用。方法:采用消化贴壁法培养SD胎鼠外胚间充质细胞,建立外胚间充质细胞的体外培养细胞模型。将生长状态良好的第三代颌突间充质细胞培养于含有20 ng/mlIGF-1、50 ml/L胎牛血清(FBS)的DMEM/F12培养基中,连续培养10d。用相差倒置显微镜观察细胞的生长和形态变化,HE染色,观察细胞形态变化。抗I型胶原单抗免疫组化染色鉴定细胞性质,碱性磷酸酶染色及活性检测。结果:原代培养的间充质细胞呈纤维样细胞生长,免疫组化染色,抗VIM为阳性,抗CK为阴性。经IGF-1培养的外胚间充质细胞,10d后在倒置显微镜下可见细胞形态发生明显变化,呈多边形,细胞紧密排列,伸出多个突起,似成骨细胞。细胞爬片HE染色,可见细胞呈不规则形态,边缘伸出多个突起。抗I型胶原单抗(Collage-I)免疫组化染色阳性,碱性磷酸酶(ALP)染色阳性,碱性磷酸酶(ALP)活性增高。结论:颌突外胚间充质细胞经胰岛素样生长因子-1(IGF-1)的诱导培养,细胞形态出现类成骨样改变,同时细胞分泌碱性磷酸酶和I型胶原,出现类似成骨细胞的功能性改变。
[Abstract]:Aim: to establish an in vitro culture model of undifferentiated extracellular mesenchymal cells from maxillary process of SD fetal rats, and to investigate the effect of insulin-like growth factor-1 (IGF-1) on differentiation of mesenchymal cells from maxillary process to osteoblasts (OB). Methods: the mesenchymal cells of SD rat embryo were cultured by digestion and adherent method, and the model of cultured cells was established in vitro. The third generation of maxillary process mesenchymal cells were cultured in DMEM / F12 medium containing 20 ng / ml IGF-1 / 50 ml / L fetal bovine serum (FBS) for 10 days. Cell growth and morphological changes were observed by phase contrast inverted microscope. Immunohistochemical staining of anti-type I collagen monoclonal antibody was used to identify cell properties, alkaline phosphatase staining and activity detection. Results: the primary cultured mesenchymal cells grew as fibroid cells, and immunohistochemical staining showed that anti-vim was positive and anti-CK was negative. After 10 days of IGF-1 culture, obvious changes of cell morphology were observed under inverted microscope, with polygon, tight arrangement of cells, multiple protrusions and osteoblast-like cells. Cell climbing slice HE staining, the cells showed irregular shape, the edge of multiple protrusions. The immunohistochemical staining of anti type I collagen monoclonal antibody (Collage-I) and alkaline phosphatase (ALP) were positive, and the activity of alkaline phosphatase (ALP) was increased. Conclusion: induced by insulin-like growth factor-1 (IGF-1), osteoblast-like changes were observed in the mesenchymal cells of the maxillary process, and alkaline phosphatase and collagen I were secreted by the cells, and the functional changes were similar to those of the osteoblasts.
【学位授予单位】：昆明医学院
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R329

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