【摘要】： 目的探討在體外誘導(dǎo)兔骨髓間充質(zhì)干細(xì)胞(Bone Marrow Mesenehymal Stem Cells,BMSCs)向上皮細(xì)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(xì)胞,體外培養(yǎng)并擴(kuò)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(xì)胞,體外培養(yǎng)擴(kuò)增,觀察其形態(tài)學(xué)變化并鑒定。第三步:采用表皮生長(zhǎng)因子(Epidermal growth factor,EGF)和上皮細(xì)胞共培養(yǎng)誘導(dǎo)法,與上皮細(xì)胞共培養(yǎng)誘導(dǎo)等方法誘導(dǎo)其向上皮細(xì)胞分化,通過形態(tài)學(xué)觀察及免疫組化方法比較各種誘導(dǎo)方法的誘導(dǎo)效率。 結(jié)果本實(shí)驗(yàn)獲得的兔骨髓間充質(zhì)干細(xì)胞增殖速度快,細(xì)胞純度高。在各組誘導(dǎo)實(shí)驗(yàn)中,經(jīng)統(tǒng)計(jì)學(xué)分析,SNK-q檢驗(yàn)比較,以表皮生長(zhǎng)因子+共培養(yǎng)聯(lián)合誘導(dǎo)組效率最高,單純共培養(yǎng)誘導(dǎo)組、表皮生長(zhǎng)因子誘導(dǎo)組次之,二組誘導(dǎo)率無(wú)顯著差異,單純以上清液誘導(dǎo)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(xì)胞。表皮生長(zhǎng)因子+上皮細(xì)胞共培養(yǎng)能有效地誘導(dǎo)其向上皮細(xì)胞細(xì)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(xué)位授予單位】：南昌大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2009
【分類號(hào)】：R329

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