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

【摘要】：目的 探討體外誘導(dǎo)犬骨髓間充質(zhì)干細(xì)胞(Bone marrow mesenchymal stemcells,BMSCs)向上皮樣細(xì)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(xì)胞,觀察其生物學(xué)特性,體外培養(yǎng)擴(kuò)增后,用上皮細(xì)胞誘導(dǎo)培養(yǎng)基誘導(dǎo)培養(yǎng),并與犬食管上皮細(xì)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(xì)胞形態(tài)的變化,培養(yǎng)第7、14天,通過(guò)免疫熒光檢測(cè)、免疫細(xì)胞化學(xué)染色和流式細(xì)胞儀鑒定上皮細(xì)胞特異性標(biāo)志物廣譜細(xì)胞角蛋白(Pan-cytokeratin,CK)和上皮膜抗原(Epithelial Membrane Antigen,EMA)的表達(dá)。結(jié)果獲得的犬BMSCs體外增殖速度快,具有多向分化潛能,細(xì)胞純度高、數(shù)量多。含20ng/ml表皮生長(zhǎng)因子(Epidermal growth factor,EGF)的無(wú)血清角化細(xì)胞培養(yǎng)基(Serum-free Keratinocyte Medium,K-SFM)能夠在體外誘導(dǎo)犬BMSCs向上皮樣細(xì)胞分化,分化程度與EGF的濃度及誘導(dǎo)時(shí)間有關(guān)。與犬EECs直接接觸共培養(yǎng),犬BMSCs可以分化為上皮樣細(xì)胞。結(jié)論本實(shí)驗(yàn)提示犬BMSCs在體外能夠向上皮樣細(xì)胞分化,有望成為組織工程食管的種子細(xì)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(xué)位授予單位】：第二軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類號(hào)】：R329.2

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本文編號(hào)：1971724