Establishment of culture system for mouse lymphatic endothelial cells adhered to ratª²tail collagen

    ZHANG Biª²Cheng1, WANG Jun1, ZHAO Yong2, GAO Jianª²Fei2, GUO Yan1, CHEN Zhengª²Tang1

    1PLA Institute of Cancer, Xinqiao Hospital, Third Military Medical University, Chongqing 400037, China, 2Department of Oncology, Wuhan General Hospital, Guangzhou Military Area Command, Wuhan 430070, China

    ¡¾Abstract¡¿ AIM: To establish a convenient, cheap and stable culture system for mouse lymphatic endothelial cells (LEC). METHODS: Mouse lymphangiomas in abdominal cavity were induced by incomplete Freund¬ðs adjuvant, then disrupted and digested to obtain LEC, which were cultured in the flask or plate previously coated with ratª²tail collagen. Expressions of LEC specific markers, VEGFRª²3 and LYVEª²1, were identified by immunofluorescence. The proliferation activity of LEC was evaluated by 3Hª²TdR incorporation efficiency and cell doubling time. The capability of LEC to form lymphatic vesselª²like structures was assessed by the in vitro lymphatic vessel formation assay. RESULTS: Mouse lymphangiomas in abdominal cavity were induced successfully by incomplete Freund¬ðs adjuvant, and more than 98% living LEC were harvested. The expressions of VEGFRª²3 and LYVEª²1 were positive in LEC. In the ratª²tail collagen coated flask or plate, LEC grew well, with 3Hª²TdR incorporation efficiency increasing obviously and cell doubling time being (42.7¡À3.2) h. In the gel formed by ratª²tail collagen, LEC could form lymphatic vesselª²like structures. CONCLUSION: Mouse LEC culture system using ratª²tail collagen as adhesive is a convenient, cheap and stable culture system. ......