Culture and evaluation of rat' s mesenteric lymphatic endothelium in vitro / 中国药理学通报

ABSTRACT

Aim: To establish the technology of isolation rat' s mesenteric lymphatics and the culture method of their endothelium and confirm the ability of synthesis nitric oxide and Factor VIII related antigens in the endothelium in vitro. Methods: With an operating microscope and microsurgical instruments, rat's mesenteric lymphatics were isolated and the endothelium were primarily cultured and subcultured. Exerting immunohistochemical method, Factor VIII related antigens and nitric oxide synthase were evaluationed. The contents of NO metabolite were measured. Results: The rat' s mesenteric lymphatics were isolated and the endothelium were Fusiform shape or polygon, the monolayer cultures displayed typical cobblestone or pavingstone morphology. Factor VIII related antigens were expressed and their positive rate was 98%. The positive rate of nitric oxide synthase was 83%. Positive reaction products were buffy around nuclear. NO metabolite was (10.26 ± 2.17) mol·L -1 (n = 7). Conclusion: The culture method we introduced was very practical, simple and reliable. The endothelium is in normal structure and function characters. It also certificated that the endothelium in vitro could express nitric oxide synthase and Factor VIII related antigens.