Objective    To optimize culture system for mouse lymphatic endothelial cells (MLEC) in vitro.     Methods    BALB/c mice benign lymphangiomas were induced by intraperitoneally injection of 0.2 ml emulsified incomplete Freund’s adjuvant (1∶1 with PBS)

twice in a 15-day interval. Then tumors were mechanically disrupted and digested to obtain MLEC that plated in dishes previously coated with gelatin

rat-tail collagen

matrigel or fibrin gel in complete medium. Cell growth conditions and formed vessel-like structures were observed by inverted phase contrast microscope

and the population doubling times were obtained in cell logarithmic growth phase.     Results    A great quantity of MLEC were isolated from the induced mouse peritoneal lymphangiomas. MLEC grow well in dishes coated with gelatin or rat-tail collagen. Cell doubling time of gelatin or rat-tail collagen group was significantly shorter than that of matrigel or fibrin gel group. The numbers of spontaneous formative vessel branches in matrigel or fibrin gel coated dishes were much higher than those in the other two groups (P<0.01).     Conclusion    Gelatin or rat-tail collagen can be used as a better base material for MLEC in vitro

furthermore

matrigel or fibrin gel is a useful model for spontaneous formation of characteristic lymphatics.