Polarized intestinal hybrid cell lines derived from primary culture: establishment and characterization.

A cell culture system has been developed that produces stable gastrointestinal (GI) polarized cell lines capable of maintaining hormone secretion. A spontaneously transformed rat mucosal epithelial cell was selected for hypoxanthine/guanine phosphoribosyltransferase deficiency and transfected with a plasmid conferring hygromycin resistance (BRIE 291 cells). Fusion of these cells with dispersed small intestinal epithelia cells resulted in hybrid cell lines that retained characteristic properties of the native GI cell more effectively than the transformed tumorigenic parental cell line. Hybrid hBRIE 380 cells are uniformly cuboidal with microvilli, contain villin, are contact inhibited, are anchorage dependent, require serum supplementation for growth, and are more sensitive to virus infection than the parental BRIE 291 cells. Fusion of BRIE 291 with dispersed pancreatic islet cells has resulted in a variety of pancreatic-hormone-producing cell lines. One of these, hybrid hBRIE 291-i2, forms confluent monolayers capable of synthesizing insulin-like immunoreactivity. These studies demonstrate that functionally polarized GI cells can be generated from primary cultures of nondividing committed epithelial cells by somatic cell hybridization and make feasible the selection and maintenance of specific GI epithelial cell types in confluent monolayer cultures. IMAGES:

本研究已开发一套细胞培养系统，可获得稳定的胃肠道（GI）极化细胞系，该细胞系能够维持激素分泌功能。研究筛选出一株携带次黄嘌呤/鸟嘌呤磷酸核糖转移酶（hypoxanthine/guanine phosphoribosyltransferase）缺陷的自发转化大鼠黏膜上皮细胞，通过携带潮霉素抗性的质粒对其进行转染，获得BRIE 291细胞。将该细胞与分散的小肠上皮细胞融合，所得到的杂交细胞系相较于亲本转化致瘤细胞系，能够更有效地保留天然胃肠道细胞的特征属性。杂交细胞hBRIE 380均一呈立方形，带有微绒毛（microvilli），表达绒毛蛋白（villin），具有接触抑制特性，锚定依赖性生长，生长需添加血清，且相较于亲本BRIE 291细胞，对病毒感染更为敏感。将BRIE 291细胞与分散的胰岛细胞融合，获得了多种产胰腺激素的细胞系。其中一株杂交细胞hBRIE 291-i2可形成汇合单层，能够合成胰岛素样免疫反应性物质。上述研究证实，通过体细胞杂交（somatic cell hybridization）技术，可从非分裂的定型上皮细胞原代培养物中获得具有功能的极化胃肠道细胞，这使得在汇合单层培养中筛选并维持特定胃肠道上皮细胞类型成为可行。IMAGES: