体外诱导兔骨髓间充质干细胞向上皮细胞分化的实验研究
发布时间：2018-10-26 11:17
【摘要】： 目的探讨在体外诱导兔骨髓间充质干细胞(Bone Marrow Mesenehymal Stem Cells,BMSCs)向上皮细胞分化的的有效方法。 方法第一步:自免阴道中提取其上皮细胞,体外培养并扩增,且观察其形态并鉴定。第二步:自兔髂骨抽取骨髓液,采用密度梯度离心法+贴壁培养法分离和提纯骨髓间充质干细胞,体外培养扩增,观察其形态学变化并鉴定。第三步:采用表皮生长因子(Epidermal growth factor,EGF)和上皮细胞共培养诱导法,与上皮细胞共培养诱导等方法诱导其向上皮细胞分化,通过形态学观察及免疫组化方法比较各种诱导方法的诱导效率。 结果本实验获得的兔骨髓间充质干细胞增殖速度快,细胞纯度高。在各组诱导实验中,经统计学分析,SNK-q检验比较,以表皮生长因子+共培养联合诱导组效率最高,单纯共培养诱导组、表皮生长因子诱导组次之,二组诱导率无显著差异,单纯以上清液诱导组效率最低。 结论密度梯度离心结合贴壁筛选能获得较纯的骨髓间充质干细胞。表皮生长因子+上皮细胞共培养能有效地诱导其向上皮细胞细胞分化。
[Abstract]:Objective to explore an effective method for inducing the differentiation of rabbit bone marrow mesenchymal stem cells (Bone Marrow Mesenehymal Stem Cells,BMSCs) into epithelial cells in vitro. Methods the first step: the epithelial cells were isolated from the vagina, cultured and amplified in vitro, and their morphology was observed and identified. The second step: bone marrow fluid was extracted from rabbit iliac bone and bone marrow mesenchymal stem cells were isolated and purified by density gradient centrifugation. The bone marrow mesenchymal stem cells were cultured and amplified in vitro. The morphological changes were observed and identified. The third step: epidermal growth factor (Epidermal growth factor,EGF) and co-culture of epithelial cells were used to induce their differentiation into epithelial cells. The induction efficiency of various induction methods was compared by morphological observation and immunohistochemical method. Results the rabbit bone marrow mesenchymal stem cells proliferated quickly and the purity of the cells was high. In the induction experiment of each group, by statistical analysis and SNK-q test, the efficiency of co-culture induced by epidermal growth factor was the highest, while that of co-culture alone and epidermal growth factor was the second, and there was no significant difference in the induction rate between the two groups. The efficiency of supernatant induction group was the lowest. Conclusion Pure bone marrow mesenchymal stem cells can be obtained by density gradient centrifugation combined with adherent screening. The co-culture of epidermal growth factor (EGF) epithelial cells can effectively induce them to differentiate into epithelial cells.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329
本文编号：2295545
[Abstract]:Objective to explore an effective method for inducing the differentiation of rabbit bone marrow mesenchymal stem cells (Bone Marrow Mesenehymal Stem Cells,BMSCs) into epithelial cells in vitro. Methods the first step: the epithelial cells were isolated from the vagina, cultured and amplified in vitro, and their morphology was observed and identified. The second step: bone marrow fluid was extracted from rabbit iliac bone and bone marrow mesenchymal stem cells were isolated and purified by density gradient centrifugation. The bone marrow mesenchymal stem cells were cultured and amplified in vitro. The morphological changes were observed and identified. The third step: epidermal growth factor (Epidermal growth factor,EGF) and co-culture of epithelial cells were used to induce their differentiation into epithelial cells. The induction efficiency of various induction methods was compared by morphological observation and immunohistochemical method. Results the rabbit bone marrow mesenchymal stem cells proliferated quickly and the purity of the cells was high. In the induction experiment of each group, by statistical analysis and SNK-q test, the efficiency of co-culture induced by epidermal growth factor was the highest, while that of co-culture alone and epidermal growth factor was the second, and there was no significant difference in the induction rate between the two groups. The efficiency of supernatant induction group was the lowest. Conclusion Pure bone marrow mesenchymal stem cells can be obtained by density gradient centrifugation combined with adherent screening. The co-culture of epidermal growth factor (EGF) epithelial cells can effectively induce them to differentiate into epithelial cells.
【学位授予单位】：南昌大学
【学位级别】：硕士
【学位授予年份】：2009
【分类号】：R329
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