论文部分内容阅读
传统的单层平面培养仍然是目前肝癌研究中最常用的体外培养方法，而这种培养方法不能形成类似体内的结构，在研究体内细胞与细胞、细胞与微环境之间的相互作用等方面存在明显的限制性．以微载体Ｃｙｔｏｄｅｘ－３为支持物，在静置状态下培养人肝癌细胞ＢＥＬ－７４０２，构建一种简单易行的人肝癌细胞三维培养模型．电子显微镜、酶活性及流式细胞仪分析表明，细胞呈多面体形，多层重叠排列呈巢状．胞间有０．５－２．０μｍ的间隙，胞间细胞膜局部突起密切接触，有桥粒形成． ９０％以上细胞存活，并维持葡萄糖的分解代谢和表达ＥＧＦ受体．胞内ＡＳＴ，ＡＬＴ及ＬＤＨ－Ｌ的活性高于平面培养．该三维结构具有体内索状型和实体型肝癌的结构特征，较之单层平面培养，更适合于研究肝癌细胞间及细胞与微环境之间的相互作用，以及用于肝癌细胞浸润、转移和抗药性机理的研究．
Traditional monolayer culture is still the most commonly used in vitro culture method for liver cancer research. However, this culture method cannot form a structure similar to that in vivo. It exists in the study of the interaction between cells and cells, cells and the microenvironment in vivo. Obvious restrictions. The human hepatocellular carcinoma cell line BEL-7402 was cultured under static conditions with the microcarrier Cytodex-3 as a support to construct a simple three-dimensional culture model of human hepatoma cells. Electron microscopy, enzymatic activity and flow cytometry analysis showed that the cells were polyhedral and multi-layered and nested. There was a gap of 0.5-2.0 μm between the cells, and the local cell membranes were in close contact with each other, resulting in the formation of desmosomes. More than 90% of cells survive and maintain glucose catabolism and expression of EGF receptors. The activity of intracellular AST, ALT and LDH-L is higher than that of planar culture. The three-dimensional structure has the structural features of cord-like and solid-type liver cancer in vivo, and is more suitable for the study of the interaction between hepatoma cells and cells and the microenvironment, and for the infiltration and metastasis of hepatocellular carcinoma cells, as compared with single-layer planar culture. Drug resistance mechanism research.