本文選題：毛細胞前體細胞 + 細胞培養(yǎng)　； 參考：《中華耳科學雜志》2016年02期

【摘要】：目的比較聯(lián)合應用嗜熱菌蛋白酶和I型膠原酶消化對耳蝸基底膜上皮細胞的分離效果。方法分離P0-3天SD大鼠基底膜,并將其分為四組,分別為:A組胰酶消化組;B組嗜熱菌蛋白酶消化組;C組I型膠原酶消化組;D組嗜熱菌蛋白酶和I型膠原酶消化組。收集各組消化的細胞進行懸浮培養(yǎng)和誘導分化,分別計數(shù)各組形成的細胞球數(shù)目,應用免疫熒光對獲得的細胞來源進行鑒定,并比較各組Cytokeratin-18陽性細胞的百分率。結(jié)果從4種方法分離得到的細胞經(jīng)培養(yǎng)后均可形成細胞球,表達干細胞標記物Nestin和細胞分裂標記物Brd U。獲得細胞大部分均表達上皮來標記物E-cadherin和cytokeratin-18,而不表達間質(zhì)細胞標志物Vimentin,經(jīng)過誘導分化后可以分化成毛細胞樣細胞。應用嗜熱菌蛋白酶和I型膠原酶消化獲得的細胞球數(shù)目顯著高于其它組(P0.05);而采用胰酶消化獲得的細胞cytokeratin-18陽性率顯著低于其它組(P0.05)。結(jié)論聯(lián)合應用嗜熱菌蛋白酶和I型膠原酶消化耳蝸基底膜上皮細胞可以顯著提高基底膜上皮細胞的分離效果,從而耳蝸前體細胞的研究提供有力研究基礎(chǔ)。
[Abstract]:Objective to compare the effects of combined digestion of thermophilic protease and collagenase I on the isolation of cochlear basement membrane epithelial cells. Methods the basement membrane of SD rats was isolated from P0-3 day and divided into four groups: group A, trypsin digestion group, group B, thermophilic protease group, group C, group I, collagenase digestion group, group D, thermophilic protease and type I collagenase digestion group, respectively. The cells digested in each group were collected for suspension culture and induced differentiation. The number of cells formed in each group was counted. The source of the cells was identified by immunofluorescence and the percentage of Cytokeratin-18 positive cells in each group was compared. Results all the cells isolated from the four methods could form cell spheres and express nestin and Brd UU, the marker of stem cell and the marker of cell division. Most of the cells expressed E-cadherin and cytokeratin-18, but not the interstitial cell marker Vimentin, which could differentiate into hair-cell-like cells after induced differentiation. The number of cell balls digested with thermophilic protease and type I collagenase was significantly higher than that of other groups (P0.05), while the positive rate of cytokeratin-18 of cells digested with trypsin was significantly lower than that of other groups (P0.05). Conclusion the combination of thermophilic protease and collagenase I can significantly improve the isolation effect of basal membrane epithelial cells of cochlea and provide an effective basis for the study of cochlear precursor cells.
【作者單位】： 第四軍醫(yī)大學第一附屬醫(yī)院耳鼻咽喉頭頸外科;
【基金】：國家自然科學基金海外重大合作項目(81120108008);國家自然科學基金(81271069)~~
【分類號】：R764

【相似文獻】

相關(guān)期刊論文 前2條

1 劉俊;孔維佳;張丹;張亞民;劉貞榮;;大鼠耳蝸感覺上皮細胞的原代培養(yǎng)及其意義[J];臨床耳鼻咽喉頭頸外科雜志;2007年01期

2 ;[J];;年期

相關(guān)會議論文 前3條

1 劉俊;趙榮祥;叢品;洪錢江;;哺乳動物內(nèi)耳再生毛細胞前體細胞來源的研究(摘要)[A];中醫(yī)耳鼻喉學術(shù)傳承與研究論文匯編[C];2009年

2 劉俊;趙榮祥;叢品;洪錢江;;哺乳動物內(nèi)耳再生毛細胞前體細胞來源的研究(摘要)[A];第15屆全國中醫(yī)耳鼻喉科學術(shù)研討和繼續(xù)教育年會論文匯編[C];2009年

3 劉俊;趙榮祥;叢品;洪錢江;;哺乳動物內(nèi)耳再生毛細胞前體細胞來源的研究[A];中華中醫(yī)藥學會耳鼻喉科分會第15屆學術(shù)交流會論文集[C];2009年

相關(guān)碩士學位論文 前1條

1 管明;骨髓間充質(zhì)干細胞體外培養(yǎng)誘導分化為內(nèi)耳毛細胞前體細胞的研究[D];浙江大學;2007年

，

本文編號：2104385