【摘要】： 目的探討在體外誘導兔骨髓間充質(zhì)干細胞(Bone Marrow Mesenehymal Stem Cells,BMSCs)向上皮細胞分化的的有效方法。 方法第一步:自免陰道中提取其上皮細胞,體外培養(yǎng)并擴增,且觀察其形態(tài)并鑒定。第二步:自兔髂骨抽取骨髓液,采用密度梯度離心法+貼壁培養(yǎng)法分離和提純骨髓間充質(zhì)干細胞,體外培養(yǎng)擴增,觀察其形態(tài)學變化并鑒定。第三步:采用表皮生長因子(Epidermal growth factor,EGF)和上皮細胞共培養(yǎng)誘導法,與上皮細胞共培養(yǎng)誘導等方法誘導其向上皮細胞分化,通過形態(tài)學觀察及免疫組化方法比較各種誘導方法的誘導效率。 結(jié)果本實驗獲得的兔骨髓間充質(zhì)干細胞增殖速度快,細胞純度高。在各組誘導實驗中,經(jīng)統(tǒng)計學分析,SNK-q檢驗比較,以表皮生長因子+共培養(yǎng)聯(lián)合誘導組效率最高,單純共培養(yǎng)誘導組、表皮生長因子誘導組次之,二組誘導率無顯著差異,單純以上清液誘導組效率最低。 結(jié)論密度梯度離心結(jié)合貼壁篩選能獲得較純的骨髓間充質(zhì)干細胞。表皮生長因子+上皮細胞共培養(yǎng)能有效地誘導其向上皮細胞細胞分化。
[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.
【學位授予單位】：南昌大學
【學位級別】：碩士
【學位授予年份】：2009
【分類號】：R329

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