人正常腹膜間皮細胞原代培養方法_《中國普外基礎與臨床雜志》

作者：

 王沛靚 ¹ , 吳曉平 ²

1.新疆醫科大學第五附屬醫院腫瘤中心（烏魯木齊 830011）;;
2.69008部隊衛生隊（烏魯木齊 830011）;

關鍵詞：

間皮細胞腹膜細胞培養技術

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目的探討人正常腹膜間皮細胞原代培養方法。
方法以胰酶和乙二胺四乙酸磁性攪拌分離人正常腹膜間皮細胞，倒置相差顯微鏡觀察細胞形態結構和大體生長過程， Calretinin免疫熒光染色鑒定間皮細胞，初步觀察傳代細胞的貼壁生長情況。
結果培養第4天細胞貼壁良好，第15天呈鋪路石狀、生長良好，1個月時細胞密集鋪滿皿壁; 腹膜間皮細胞Calretinin表達都呈陽性; 傳代腹膜間皮細胞數量顯著減少。
結論磁性攪拌酶分離培養可得到數量多、純度高的原代人正常腹膜間皮細胞，能夠滿足一般實驗要求。

引用本文： 王沛靚,吳曉平. 人正常腹膜間皮細胞原代培養方法. 中國普外基礎與臨床雜志, 2010, 17(1): 57-59. doi: 復制

1.	劉洪斌, 李東華, 郭世鐸. 腹腔粘連形成機制及治療研究進展 [J]. 中國中西醫結合外科雜志, 2005; 11(1): 8486.
2.	張曉國, 潘定宇, 史海安, 等. 切口下腹膜粘連的實驗觀察及其防范 [J]. 中國普外基礎與臨床雜志, 2002; 9(6): 401.
3.	Peng YM, Liu H, Liu FY, et al. Troglitazone inhibits synthesis of transforming growth factor b1 and reduces matrix production in human peritoneal mesothelial cells [J]. Nephrology, 2006; 11(6): 516523.
4.	Stylianou E, Jenner LA, Davies M, et al. Isolation, culture and characterization of human peritoneal mesothelial cells [J]. Kidney Int, 1990; 37(6): 15631570.
5.	程永波, 房殿春, 郭立萍, 等. 一種簡便的人正常胃黏膜上皮細胞原代培養方法 [J]. 胃腸病學, 2007; 12(1): 3135.
6.	Haslinger B, Kleemann R, Toet KH, et al. Simvastatin suppresses tissue factor expression and increases fibrinolytic activity in tumor necrosis factoractivated human peritoneal mesothelial cells [J]. Kidney Int, 2003; 63(6): 20652074.
7.	Yang AH, Huang SW, Chen JY, et al. Leptin augments myofibroblastic conversion and fibrogenic activity of human peritoneal sothelial cells: A functional implication for peritoneal fibrosis [J]. Nephrol Dial Transplant, 2007; 22(3): 756762.
8.	樊敏, 劉伏友, 段紹斌, 等. 改良法培養鼠腹膜間皮細胞 [J]. 湖南醫科大學學報, 2002; 27(6): 542544.
9.	戴宏, 劉先英, 張伯科, 等. 腹膜透析流出液中人腹膜間皮細胞的培養 [J]. 安徽醫科大學學報, 2008; 43(4): 464466.
10.	Noh H, Ha H, Yu MR, et al. Angiotensin Ⅱ mediates high glucose induced TGβ21 and fibroncetin up regulation in HPMC through reaction oxygen species [J]. Perit Dial Int, 2005; 25(1): 3847.
11.	Yang X, Ye RG, Yong QY, et al. CD40 ligand expression on macrophages during peritonitis in continuous ambulatory peritoneal dialysis patients [J]. Adv Perit Dial, 2000; 16 (2): 213 225.
12.	Tee MM, Tesch GH, NikolicPaterson DJ, et al. Human peritoneal mesothelial cells isolated from spent dialysate fluid maintain contaminating macrophages via production of macrophage colony stimulating factor [J]. Nephrology, 2007; 12(2): 160165.
13.	Stadlmann S, Pollheimer J, Renner K, et al. Response of human peritoneal mesothelial cells to inflammatory injury is regulated by interleukin1beta and tumor necrosis factoralpha [J]. Wound Repair Regen, 2006; 14(2): 187194.
14.	Xu ZG, Kim KS, Park HC, et al. High glucose activates the p38 MAPK pathway in cultured human peritoneal mesothelial cells [J]. Kidney Int, 2003; 63(3): 958968.

1. 劉洪斌, 李東華, 郭世鐸. 腹腔粘連形成機制及治療研究進展 [J]. 中國中西醫結合外科雜志, 2005; 11(1): 8486.
2. 張曉國, 潘定宇, 史海安, 等. 切口下腹膜粘連的實驗觀察及其防范 [J]. 中國普外基礎與臨床雜志, 2002; 9(6): 401.
3. Peng YM, Liu H, Liu FY, et al. Troglitazone inhibits synthesis of transforming growth factor b1 and reduces matrix production in human peritoneal mesothelial cells [J]. Nephrology, 2006; 11(6): 516523.
4. Stylianou E, Jenner LA, Davies M, et al. Isolation, culture and characterization of human peritoneal mesothelial cells [J]. Kidney Int, 1990; 37(6): 15631570.
5. 程永波, 房殿春, 郭立萍, 等. 一種簡便的人正常胃黏膜上皮細胞原代培養方法 [J]. 胃腸病學, 2007; 12(1): 3135.
6. Haslinger B, Kleemann R, Toet KH, et al. Simvastatin suppresses tissue factor expression and increases fibrinolytic activity in tumor necrosis factoractivated human peritoneal mesothelial cells [J]. Kidney Int, 2003; 63(6): 20652074.
7. Yang AH, Huang SW, Chen JY, et al. Leptin augments myofibroblastic conversion and fibrogenic activity of human peritoneal sothelial cells: A functional implication for peritoneal fibrosis [J]. Nephrol Dial Transplant, 2007; 22(3): 756762.
8. 樊敏, 劉伏友, 段紹斌, 等. 改良法培養鼠腹膜間皮細胞 [J]. 湖南醫科大學學報, 2002; 27(6): 542544.
9. 戴宏, 劉先英, 張伯科, 等. 腹膜透析流出液中人腹膜間皮細胞的培養 [J]. 安徽醫科大學學報, 2008; 43(4): 464466.
10. Noh H, Ha H, Yu MR, et al. Angiotensin Ⅱ mediates high glucose induced TGβ21 and fibroncetin up regulation in HPMC through reaction oxygen species [J]. Perit Dial Int, 2005; 25(1): 3847.
11. Yang X, Ye RG, Yong QY, et al. CD40 ligand expression on macrophages during peritonitis in continuous ambulatory peritoneal dialysis patients [J]. Adv Perit Dial, 2000; 16 (2): 213 225.
12. Tee MM, Tesch GH, NikolicPaterson DJ, et al. Human peritoneal mesothelial cells isolated from spent dialysate fluid maintain contaminating macrophages via production of macrophage colony stimulating factor [J]. Nephrology, 2007; 12(2): 160165.
13. Stadlmann S, Pollheimer J, Renner K, et al. Response of human peritoneal mesothelial cells to inflammatory injury is regulated by interleukin1beta and tumor necrosis factoralpha [J]. Wound Repair Regen, 2006; 14(2): 187194.
14. Xu ZG, Kim KS, Park HC, et al. High glucose activates the p38 MAPK pathway in cultured human peritoneal mesothelial cells [J]. Kidney Int, 2003; 63(3): 958968.

《中國普外基礎與臨床雜志》

人正常腹膜間皮細胞原代培養方法

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

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《中國普外基礎與臨床雜志》

人正常腹膜間皮細胞原代培養方法

摘要 全文 圖表 視頻 參考文獻 施引文獻 補充材料

Format

Content

摘要全文圖表視頻參考文獻施引文獻補充材料