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

目的  探討人正常腹膜間皮細胞原代培養(yǎng)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(tài)結(jié)構(gòu)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(jié)果  培養(yǎng)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(shù)量顯著減少。
結(jié)論  磁性攪拌酶分離培養(yǎng)可得到數(shù)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(yǎng)方法. 中國普外基礎(chǔ)與臨床雜志, 2010, 17(1): 57-59. doi: 復(fù)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(yī)結(jié)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(chǔ)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(yǎng)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(yǎng)鼠腹膜間皮細胞 [J]. 湖南醫(yī)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(yǎng) [J]. 安徽醫(yī)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. 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(yī)結(jié)合外科雜志, 2005; 11(1): 8486.
  2. 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(chǔ)與臨床雜志, 2002; 9(6): 401.
  3. 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. 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. 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(yǎng)方法 [J]. 胃腸病學, 2007; 12(1): 3135.
  6. 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. 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. 8. 樊敏, 劉伏友, 段紹斌, 等. 改良法培養(yǎng)鼠腹膜間皮細胞 [J]. 湖南醫(yī)科大學學報, 2002; 27(6): 542544.
  9. 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(yǎng) [J]. 安徽醫(yī)科大學學報, 2008; 43(4): 464466.
  10. 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. 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. 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. 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. 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.