本文關(guān)鍵詞：人臍帶間充質(zhì)干細(xì)胞分化胰島樣細(xì)胞過(guò)程中胰島素和巢蛋白的表達(dá)　出處：《中國(guó)組織工程研究》2016年50期 　論文類(lèi)型：期刊論文

  更多相關(guān)文章： 臍帶 間質(zhì)干細(xì)胞 胰島素分泌細(xì)胞 胰島素 組織工程 干細(xì)胞 臍帶臍血干細(xì)胞 臍帶間充質(zhì)干細(xì)胞 胰島樣細(xì)胞 誘導(dǎo)分化 巢蛋白

【摘要】：背景:國(guó)內(nèi)外研究證實(shí)臍帶間充質(zhì)干細(xì)胞可定向誘導(dǎo)分化為胰島樣細(xì)胞,但分化過(guò)程中胰島素與巢蛋白表達(dá)的變化等相關(guān)研究甚少。目的:觀(guān)察人臍帶間充質(zhì)干細(xì)胞向胰島素樣細(xì)胞分化過(guò)程中胰島素與巢蛋白表達(dá)的變化。方法:采用UltraCULTURE培養(yǎng)基在體外培養(yǎng)人臍帶間充質(zhì)干細(xì)胞,取培養(yǎng)的第3代細(xì)胞,按照兩階段誘導(dǎo)方案將其定向誘導(dǎo)分化為胰島樣細(xì)胞,第1階段:在UltraCULTURE培養(yǎng)基中加入4 nmol/L活化素A,25μg/L表皮生長(zhǎng)因子,100μg/Lβ-神經(jīng)生長(zhǎng)因子,10 mmol/L尼克酰胺,誘導(dǎo)培養(yǎng)至14 d。第2階段:在UltraCULTURE培養(yǎng)基中加入1%胰島素-轉(zhuǎn)鐵蛋白-硒,10 mmol/L尼克酰胺,10μg/L堿性成纖維細(xì)胞生長(zhǎng)因子,繼續(xù)誘導(dǎo)培養(yǎng)14 d。在誘導(dǎo)分化過(guò)程中,采用流式細(xì)胞技術(shù)檢測(cè)分化細(xì)胞巢蛋白及胰島素的表達(dá),雙硫腙染色鑒定胰島樣細(xì)胞團(tuán)中鋅離子的表達(dá)。結(jié)果與結(jié)論:(1)誘導(dǎo)分化為胰島樣細(xì)胞過(guò)程中胰島素水平逐漸增高,至28 d達(dá)到較高水平,未誘導(dǎo)組胰島素表達(dá)呈陰性;(2)誘導(dǎo)分化第14天,巢蛋白表達(dá)水平達(dá)到最大,隨著誘導(dǎo)時(shí)間延長(zhǎng)表達(dá)水平逐漸下降;(3)臍帶間充質(zhì)干細(xì)胞誘導(dǎo)28 d形成胰島樣細(xì)胞團(tuán),雙硫腙染色陽(yáng)性;(4)實(shí)驗(yàn)將臍帶間充質(zhì)干細(xì)胞成功誘導(dǎo)分化為胰島樣細(xì)胞,并且隨著定向分化細(xì)胞的變化,細(xì)胞胰島素與巢蛋白表達(dá)水平也出現(xiàn)相應(yīng)的變化。
[Abstract]:Background: domestic and foreign studies confirm that umbilical cord mesenchymal stem cells can be induced to differentiate into islet like cells, but there is little research about the change of insulin and nestin expression during differentiation. Objective: To observe the changes in the expression of insulin and nestin in human umbilical cord mesenchymal stem cells (MSCs) to insulin-like cells. Methods: UltraCULTURE cultured human umbilical cord mesenchymal stem cells in vitro culture medium, cultured the cells of the third generation, according to the two phase of induced directionally induced BMSCs into islet like cells, the first stage: in 4 nmol/L activin A added UltraCULTURE medium, 25 g/L of epidermal growth factor, 100 g/L nerve growth factor beta, 10 mmol/L nicotinamide, cultured to 14 d. The second stage: after adding 1% insulin transferrin selenium UltraCULTURE medium, 10 mmol/L nicotinamide, 10 g/L basic fibroblast growth factor, continued to be cultured 14 d. During induction and differentiation, the expression of nestin and insulin in differentiated cells was detected by flow cytometry, and the expression of zinc ions in islet like cell clusters was identified by Dithizone staining. Results and conclusion: (1) differentiation into islet like cells in insulin levels increased to 28 d reached a high level, non induced expression of insulin group were negative; (2) fourteenth days of differentiation, the expression of nestin reached the maximum with time of induction was decreased; (3) umbilical cord mesenchymal stem cells induced by 28 d into islet like cell clusters. Dithizone staining; (4) the umbilical cord mesenchymal stem cells were successfully induced to differentiate into islet like cells, and with the change of cell differentiation, expression of nestin and insulin levels also appeared the corresponding changes.
【作者單位】： 北京大學(xué)航天臨床醫(yī)學(xué)院血液內(nèi)分泌科;中國(guó)中醫(yī)科學(xué)院中藥所藥理室;
【基金】：航天中心醫(yī)院科研基金(KY-201003),課題名稱(chēng):人骨髓間充質(zhì)干細(xì)胞向胰島細(xì)胞定向分化的研究~~
【分類(lèi)號(hào)】：R329.2
【正文快照】： 文章快速閱讀:利用體外微環(huán)境誘導(dǎo)人臍帶間充質(zhì)干細(xì)胞定向分化為胰島樣細(xì)胞的過(guò)程人臍帶間充質(zhì)干細(xì)胞的體外分離、培養(yǎng)與鑒定細(xì)胞形態(tài)學(xué)觀(guān)察、流式細(xì)胞儀檢測(cè)表面表型、成骨成脂誘導(dǎo)分化步驟1誘導(dǎo)第1階段臍帶間充質(zhì)干細(xì)胞向神經(jīng)樣細(xì)胞分化步驟2誘導(dǎo)第2階段細(xì)胞形態(tài)學(xué)觀(guān)察、流

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