• 1.四川大學(xué)華西醫(yī)院衛(wèi)生部移植工程和移植免疫重點(diǎn)實(shí)驗(yàn)室（成都610041）;;
  • 2.四川大學(xué)華西醫(yī)院肝膽胰外科;;
  • 3.四川大學(xué)華西醫(yī)院病理科;

目的探索人膽囊上皮細(xì)胞的最佳分離方法和適宜的體外生長(zhǎng)條件，為深入研究膽囊的生理功能和相關(guān)疾病的病理機(jī)理奠定基礎(chǔ)。方法用Ⅳ型膠原酶消化及鈍性刮離法分離膽囊上皮細(xì)胞，兩步貼壁法純化。比較層粘連蛋白、多聚賴(lài)氨酸、纖維連接蛋白等不同培養(yǎng)基質(zhì)對(duì)傳代細(xì)胞生長(zhǎng)的影響。光鏡及電鏡下觀察細(xì)胞的形態(tài)和超微結(jié)構(gòu)。結(jié)果每個(gè)膽囊可分離得到（1～5）×107個(gè)膽囊上皮細(xì)胞，細(xì)胞活性可達(dá)90%。細(xì)胞呈典型的扁平多角狀、柱狀形態(tài)，片狀貼壁生長(zhǎng)，上皮細(xì)胞特異性抗原CK19表達(dá)陽(yáng)性。結(jié)論Ⅳ型膠原酶消化法加鈍性刮剝分離法和兩步貼壁法可成功分離高活性、高純度的人膽囊上皮細(xì)胞； 纖維連接蛋白包被的培養(yǎng)器皿和含20%小牛血清及10 ng/ml hEGF的DMEM培養(yǎng)基有利于膽囊上皮細(xì)胞的體外培養(yǎng)。

引用本文： 李勝富,潘光棟,劉瑾,熊先澤,劉江文,張杰,步宏,程南生. 體外培養(yǎng)人膽囊上皮細(xì)胞的形態(tài)學(xué)及培養(yǎng)條件的優(yōu)化. 中國(guó)普外基礎(chǔ)與臨床雜志, 2003, 10(5): 459-462. doi: 復(fù)制

1. Oda D, Lee SP, Hayashi A. Longterm culture and partial characterization of dog gallbladder epithelial cells [Ｊ]. Lab Invest， 1991; 64(5)∶682.
2. Luciano L, Reale E. Brush cells of the mouse gallbladder. A correlative light and electronmicroscopical study [Ｊ]. Cell Tissue Res，1990; 262(2)∶339.
3. O’Brien JA, James PS, Torre M. Primary cultures of guineapig gallbladder epithelial cells suitable for patchclamp studies [Ｊ]. Exp Physiol， 1991; 76(5)∶807.
4. Plevris JN, Walker SW, Harrison DJ, et al. Primary culture of bovine gall bladder epithelial cells [Ｊ]. Gut， 1993; 34(11)∶1612.
5. Kozuka S, Tsubone N, Yasui A, et al. Relation of adenoma to carcinoma in the gallbladder [Ｊ]. Cancer， 1982; 50(10)∶2226.
6. 張兆祥, 尹為華, 朱忠堯， 等. 膽囊腺瘤和腺瘤癌變 [Ｊ]. 中華外科雜志， 1994; 32(4)∶400.
  1. 1. Oda D, Lee SP, Hayashi A. Longterm culture and partial characterization of dog gallbladder epithelial cells [Ｊ]. Lab Invest， 1991; 64(5)∶682.
  2. 2. Luciano L, Reale E. Brush cells of the mouse gallbladder. A correlative light and electronmicroscopical study [Ｊ]. Cell Tissue Res，1990; 262(2)∶339.
  3. 3. O’Brien JA, James PS, Torre M. Primary cultures of guineapig gallbladder epithelial cells suitable for patchclamp studies [Ｊ]. Exp Physiol， 1991; 76(5)∶807.
  4. 4. Plevris JN, Walker SW, Harrison DJ, et al. Primary culture of bovine gall bladder epithelial cells [Ｊ]. Gut， 1993; 34(11)∶1612.
  5. 5. Kozuka S, Tsubone N, Yasui A, et al. Relation of adenoma to carcinoma in the gallbladder [Ｊ]. Cancer， 1982; 50(10)∶2226.
  6. 6. 張兆祥, 尹為華, 朱忠堯， 等. 膽囊腺瘤和腺瘤癌變 [Ｊ]. 中華外科雜志， 1994; 32(4)∶400.