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  本文選題：組織工程 + 干細胞��； 參考：《第二軍醫(yī)大學》2006年碩士論文

【摘要】：目的 探討體外誘導犬骨髓間充質(zhì)干細胞(Bone marrow mesenchymal stemcells,BMSCs)向上皮樣細胞分化的可能性和基本條件。方法采用密度梯度離心和貼壁培養(yǎng)相結(jié)合的方法分離提純?nèi)撬栝g充質(zhì)干細胞,觀察其生物學特性,體外培養(yǎng)擴增后,用上皮細胞誘導培養(yǎng)基誘導培養(yǎng),并與犬食管上皮細胞(Esophageal epithelial cells,EECs)直接接觸共培養(yǎng),每日在相差顯微鏡下觀察細胞形態(tài)的變化,培養(yǎng)第7、14天,通過免疫熒光檢測、免疫細胞化學染色和流式細胞儀鑒定上皮細胞特異性標志物廣譜細胞角蛋白(Pan-cytokeratin,CK)和上皮膜抗原(Epithelial Membrane Antigen,EMA)的表達。結(jié)果獲得的犬BMSCs體外增殖速度快,具有多向分化潛能,細胞純度高、數(shù)量多。含20ng/ml表皮生長因子(Epidermal growth factor,EGF)的無血清角化細胞培養(yǎng)基(Serum-free Keratinocyte Medium,K-SFM)能夠在體外誘導犬BMSCs向上皮樣細胞分化,分化程度與EGF的濃度及誘導時間有關。與犬EECs直接接觸共培養(yǎng),犬BMSCs可以分化為上皮樣細胞。結(jié)論本實驗提示犬BMSCs在體外能夠向上皮樣細胞分化,有望成為組織工程食管的種子細胞。
[Abstract]:Objective to investigate the possibility and basic conditions of inducing the differentiation of canine bone marrow mesenchymal stem cells (BMSCs) into epithelioid cells in vitro. Methods Canine bone marrow mesenchymal stem cells were isolated and purified by density gradient centrifugation and adherent culture. The cells were cultured directly with esophageal epithelial cells (Esophageal epithelial cells). The morphological changes of the cells were observed under phase contrast microscope every day. The cells were cultured for 714 days and were detected by immunofluorescence. Immunocytochemical staining and flow cytometry were used to identify the expression of Pan-cytokeratin CKand Epithelial Membrane antigenase (EMA), a specific marker of epithelial cells. Results the canine BMSCs had the advantages of high proliferation rate, multidirectional differentiation potential, high cell purity and large number of cells. Serum-free Keratinocyte Medium-K-SFM, a serum-free Keratinocyte medium containing 20ng/ml epidermal growth factor (EGF), could induce canine BMSCs to differentiate into epithelioid cells in vitro, and the differentiation degree was related to the concentration and time of EGF. Canine BMSCs can differentiate into epithelioid cells by direct contact with canine EECs. Conclusion this study suggests that canine BMSCs can differentiate into epithelioid cells in vitro and may become seed cells of tissue engineering esophagus.
【學位授予單位】：第二軍醫(yī)大學
【學位級別】：碩士
【學位授予年份】：2006
【分類號】：R329.2

【參考文獻】

相關期刊論文 前3條

1 劉曉丹,郭子寬,李秀森,張雙喜,毛寧;人骨髓間充質(zhì)干細胞分離與培養(yǎng)方法的建立[J];軍事醫(yī)學科學院院刊;2000年04期

2 方利君,付小兵,王玉新,孫同柱,程飚;體外誘導骨髓間充質(zhì)干細胞分化為上皮細胞的研究[J];中華實驗外科雜志;2004年02期

3 李海紅,付小兵,周崗,費沛,陳偉,白曉東,蔡存良,孫同柱;人骨髓間充質(zhì)干細胞與汗腺細胞共同培養(yǎng)誘導細胞表型轉(zhuǎn)化的初步研究[J];中華醫(yī)學雜志;2005年27期

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本文編號：1971724