小鼠淋巴管内皮细胞体外不同培养体系的比较

目的 探讨最适合小鼠淋巴管内皮细胞(mouse lymphatic endothelial cell,MLEC)生长的体系.方法 小鼠腹腔注射乳化的不完全弗式佐剂(15 d后再注射1次),诱导形成良性淋巴管瘤.消化法分离MLEC,分别置于明胶、鼠尾胶、基质胶或纤维蛋白凝胶包被的培养板中生长,流式细胞术对MLEC进行鉴定,倒置相差显微镜观察细胞生长状态和形成管样结构的差异,并计算细胞群体倍增时间.结果 从淋巴管瘤中分离得到MLEC.MLEC在明胶、鼠尾胶支持物上生长良好,其群体倍增时间均短于在基质胶和纤维蛋白凝胶上生长的MLEC.MLEC在4种支持物上均可以形成管样结构,在基质胶和纤维蛋白凝胶上形成的管分支数明显高于其他组(P＜0.01).结论 明胶和鼠尾胶是MLEC生长的较好支持物,而基质胶和纤维蛋白凝胶是体外淋巴管形成的较好模型.

Comparison of different culture systems for mouse lymphatic endothelial cells in vitro

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.