腱组织提取物对大鼠脂肪来源间充质干细胞增殖和向肌腱细胞分化的影响
发布时间：2018-11-20 10:23
【摘要】：研究目的 探讨腱组织提取物对ADSC增殖和向肌腱细胞方向分化的影响,从而为肌腱组织工程及其最终应用于临床提供理论依据。 研究方法 从3周龄SD大鼠腹股沟脂肪垫分离出脂肪组织,通过Ⅰ型胶原酶消化法分离干细胞,接种于含有胎牛血清的DMEM培养基中分离传代。每日以倒置显微镜观察细胞形态及生长情况,MTT法绘制ADSC生长曲线。经鉴定后,取第三代ADSC分成实验组和对照组2组,利用腱组织提取物处理实验组ADSC, Hank's液处理对照组细胞后继续培养观察。采用MTT比色法进行增殖活性比较,Ⅰ型胶原免疫细胞化学染色检测成肌腱分化情况以及RT-PCR检测肌腱发育相关基因(CollagenⅠ、CollagenⅢ基因)的表达。 研究结果 原代细胞接种4-6小时后即可见沉降贴壁,7-8天左右可见大量增殖,按一定方向性排列,呈漩涡状或束状,传代后细胞增殖迅速,生长稳定。腱组织提取物诱导后,MTT实验显示ADSC增殖活性并未发生明显改变,与对照组间没有统计学差异(p0.05)。行Ⅰ型胶原免疫细胞化学染色显示：实验组为阳性,对照组阴性；RT-PCR检测显示肌腱发育相关基因(CollagenⅠ、CollagenⅢ基因)表达明显增加(p0.01)。 研究结论 1.腱组织提取物不能改变大鼠ADSC的增殖活性 2.腱组织提取物可以促进大鼠ADSC向肌腱细胞方向分化
[Abstract]:Objective to investigate the effect of tendon tissue extract on the proliferation and differentiation of ADSC in the direction of tendon cells, so as to provide theoretical basis for tendon tissue engineering and its application in clinical practice. Methods Adipose tissue was isolated from the inguinal fat pad of 3-week-old SD rats. Stem cells were isolated by type I collagenase digestion method. The stem cells were isolated and subcultured in DMEM medium containing fetal bovine serum. The morphology and growth of cells were observed by inverted microscope daily, and ADSC growth curve was drawn by MTT method. After identification, the third generation of ADSC was divided into two groups: the experimental group and the control group. The cells of the control group were treated with the extract of tendon tissue and treated with the solution of ADSC, Hank's. The proliferative activity was compared by MTT colorimetry, the differentiation of tendon was detected by type I collagen immunocytochemical staining, and the expression of Collagen 鈪，
本文编号：2344652
[Abstract]:Objective to investigate the effect of tendon tissue extract on the proliferation and differentiation of ADSC in the direction of tendon cells, so as to provide theoretical basis for tendon tissue engineering and its application in clinical practice. Methods Adipose tissue was isolated from the inguinal fat pad of 3-week-old SD rats. Stem cells were isolated by type I collagenase digestion method. The stem cells were isolated and subcultured in DMEM medium containing fetal bovine serum. The morphology and growth of cells were observed by inverted microscope daily, and ADSC growth curve was drawn by MTT method. After identification, the third generation of ADSC was divided into two groups: the experimental group and the control group. The cells of the control group were treated with the extract of tendon tissue and treated with the solution of ADSC, Hank's. The proliferative activity was compared by MTT colorimetry, the differentiation of tendon was detected by type I collagen immunocytochemical staining, and the expression of Collagen 鈪，
本文编号：2344652
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