Method for Primary Culture of Normal Human Peritoneal Mesothelial Cells_CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Authors：

 WANG Peiliang ¹ , WU Xiaoping ²

1.Center of Oncology, The Fifth Affiliated Hospital of Xingjiang Medical University, Urumqi 830011, China;;
2.Health Team of 69008 Unit, Urumqi 830011, China;

Corresponding author：

WANG Peiliang , Email: wpl123456@126.com

Keywords：

Mesothelial cell; Peritoneum; Cell culture technique

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Objective To develop a reliable method for primary culture of normal human peritoneal mesothelial cells. Methods Human peritoneal mesothelial cells were dissociated by a mixture of pancreatin and ethylene diamine tetraacetic acid with a magnetic puddler. Inverted phase contrast microscope was used to observe the morphological structures of cells, approximate process of growth. Calretinin was used to identify the mesothelial cells.
Results On the 4th d of culture, mesothelial cells adhered to the culture dish. After day 14, mesothelial cells confluenced gradually and grew well like the slabstone. Calretinin was positively expressed by mesothelial cells after 5 d of cultivation. The mesothelial cell population of subculture was less than that of the primary culture. Conclusion A reliable method for primary culture of normal human peritoneal mesothelial cells has been successfully developed, by which sufficient amount of highly purified normal human peritoneal mesothelial cells can be obtained.

Citation： WANG Peiliang ,WU Xiaoping. Method for Primary Culture of Normal Human Peritoneal Mesothelial Cells. CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY, 2010, 17(1): 57-59. doi: Copy

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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, 刘伏友, 段绍斌, 等. 改良法培养鼠腹膜间皮细胞 [J]. 湖南医科大学学报, 2002; 27(6): 542544.
9.	戴宏, 刘先英, 张伯科, 等. 腹膜透析流出液中人腹膜间皮细胞的培养 [J]. 安徽医科大学学报, 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. 刘洪斌, 李东华, 郭世铎. 腹腔粘连形成机制及治疗研究进展 [J]. 中国中西医结合外科杂志, 2005; 11(1): 8486.
2. 张晓国, 潘定宇, 史海安, 等. 切口下腹膜粘连的实验观察及其防范 [J]. 中国普外基础与临床杂志, 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. 一种简便的人正常胃黏膜上皮细胞原代培养方法 [J]. 胃肠病学, 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, 刘伏友, 段绍斌, 等. 改良法培养鼠腹膜间皮细胞 [J]. 湖南医科大学学报, 2002; 27(6): 542544.
9. 戴宏, 刘先英, 张伯科, 等. 腹膜透析流出液中人腹膜间皮细胞的培养 [J]. 安徽医科大学学报, 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.

CHINESE JOURNAL OF BASES AND CLINICS IN GENERAL SURGERY

Method for Primary Culture of Normal Human Peritoneal Mesothelial Cells

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