淋巴管内皮细胞体外培养模型的建立

Establishment of Culture Model of Lymphatic Endothelial Cells in Vitro

目的建立淋巴管内皮细胞培养模型,为淋巴管生成及肿瘤淋巴管转移等方面的研究提供细胞生物学基础。方法用胶原酶灌注消化的方法,从猪胸导管中分离出淋巴管内皮细胞进行原代及传代培养,应用免疫细胞化学法对培养的细胞进行鉴定。结果原代细胞培养3～5h后,大部分活力较高的内皮细胞已开始贴壁,12～24h后,内皮细胞己铺展形成由数个细胞组成的细胞群,1～2w后,细胞呈生长密集状并汇合形成单层,在倒置相差显微镜下,呈特征性"鹅卵石状"或"铺路石状"镶嵌排列生长。免疫荧光化学检测表明,淋巴管内皮细胞表达Ⅷ因子和血管内皮生长因子受体-3。结论该模型具有体内淋巴管内皮细胞的生物学特性,可用于淋巴管生成及肿瘤淋巴管转移等方面的体外研究。

Objective To establish cultured models of lymphatic endothelial cells in vitro for study of lymphangiogenesis and lymphatic metastasis of tumor. Methods Lymphatic endothelial cells were isolated from pig thoracic duct, primary cultures and subculture were finished by collagenase digestion method, immunocytochemstry was used in identification of their biological characterization. Results Many endothelial cells were observed with higher vigor and adherance to the wall after incubation for 3-5h, and spreaded to form cell masses for 12-24h, and grew densely and converged into single layer which characteristically arranged to cobble-stone shape after incubation for 1-2 weeks. The expressions of factor Ⅷ and vascular endothelial growth factor receptor-3 were positive in lymphatic endothelial cells. Conclusions The cultured cells have the biological property and are useful for study of lymphangiogenesis and lymphatic metastasis of tumor in vitro.