人小肠癌腹水转移细胞的无血清建系培养

ESTABLISHMENT AND CHARACTERIZATION OF A SERUM-FREE HUMAN CELL LINE HICSF

人小肠癌腹水转移细胞系ＨＩＣ的无血清培养亚系经过３ａ的传代培养建立成功，命名为ＨＩＣＳＦ。该细胞仍保留其母系ＨＩＣ一些属性：亚三倍体组型，众数５３条；增殖活跃，倍增时间１６ｈ；异体动物移植致瘤，具有活跃的血管生成行为和较高的转移行为。由于培养条件的变化，该细胞也获得了一些新的特征，ＨＩＣＳＦ细胞属弱贴附细胞，形态多变。贴附细胞主要靠树枝状突起贴附于培养介质，突起内微管、微丝相对较多，细胞骨架总体上呈无序排列。ＳＤＳ－ＰＡＧＥ电泳表明ＨＩＣＳＦ细胞分泌多种蛋白质进入培养上清中，培养上清的层析分离表明，至少存在分子质量分别为９０，７０，４５和１０ｋｕ４种促生长活性因子，同时也存在分子质量分别为６０和３０ｋｕ２种生长抑制活性因子。这些生长因子共同调控着ＨＩＣＳＦ细胞的自主生长。

A serum-free subline, HICSF, has been established from the human intestinal carcinoma metastatic ascites cell line (HIC). The spindle-like polygonal cells weakly adhere to flasks via dentritic protrusions which are sensitive to environmental changes such as temperature, mechanical disturbance, etc. The cells grow rapidly with a population doubling time of 16h. The chromosome make-up is hyperdiploid with a common number of 53. Transplantation of the cells into Balb/C athymic nude mice resulted in the formation of tumors at the inoculation sites and lung metasases, with an efficiency of 5/6 and 3/5, respectively. Compared with its parental line HIC, HICSF cells displayed different morphology and cytoskeleton patterns, prolonged population doubling time and S-phase, more concentrated chromosome distribution and different glycosylation at cell surface. The differences maybe resulted from the difference of culture conditions. The conditioned medium of HICSF cells stimulated the proliferation of themselves, suggesting the existence of autocrine growth factors. Chromatography of the conditioned media revealed four growth-stimulating activities with MW of 90, 70, 45 and 10ku, respectively; and two growth-inhibitory activities with MW of 60 and 30ku,respectively. The results suggested that the co -operation of these autocrine growth factors regulate the autonomous growth of the HICSF cells.