Pang G, Buret A, O'Loughlin E, Smith A, Batey R, Clancy R
Faculty of Medicine and Australian Institute of Mucosal Immunology, University of Newcastle, New South Wales, Australia.
Gastroenterology. 1996 Jul;111(1):8-18. doi: 10.1053/gast.1996.v111.pm8698229.
BACKGROUND & AIMS: Epithelial cell cultures can be used for the study of epithelial cell biology, although human small intestinal cultures have not been available to date. The aim of this study was to characterize three cell lines derived from normal human duodenum.
Cells were cultured from tissue fragments obtained from endoscopic biopsy specimens and characterized with respect to morphology and cytokine gene expression and for the presence of vectorial transport.
All cell lines grew as polarized continuous monolayers and were mostly cuboidal in shape but were not immortalized. Cells showed junctional complexes and sparse microvilli. All cell lines showed cytokeratins and mucin antigen but not chromagranin and messenger RNA for epidermal growth factor, interleukin 6, and vascular cell adhesion molecule 1. Disaccharidase activities were low and correlated with the low proportion of cells (1%-10%), showing positive immunocytochemistry for sucrase. Monolayer resistance varied from 30 to 200 ohms. One monolayer (BN) consistently showed secretion in response to forskolin (10 micromol/L), which could b inhibited by chloride-free buffer and apical addition of the chloride channel blocker diphenylamine decarboxylate. No monolayer had evidence of glucose transport.
These three nonimmortalized lines show morphological, phenotypic, and transport characteristics of crypt-like intestinal epithelial cells. The pattern of messenger RNA expression suggests a growth-promoting and immunomodulatory role.
上皮细胞培养可用于上皮细胞生物学研究，尽管目前尚无人类小肠培养物。本研究的目的是对源自正常人十二指肠的三种细胞系进行特性分析。
从内镜活检标本获取的组织碎片中培养细胞，并对其形态、细胞因子基因表达以及是否存在向量转运进行特性分析。
所有细胞系均以极化连续单层生长，大多呈立方形，但未永生化。细胞显示出连接复合体和稀疏的微绒毛。所有细胞系均显示细胞角蛋白和黏蛋白抗原，但不显示嗜铬粒蛋白以及表皮生长因子、白细胞介素6和血管细胞黏附分子1的信使核糖核酸。双糖酶活性较低，且与蔗糖酶免疫细胞化学呈阳性的细胞比例较低（1%-10%）相关。单层电阻在30至200欧姆之间变化。一个单层（BN）对福司可林（10微摩尔/升）持续显示出分泌反应，该反应可被无氯缓冲液和顶端添加的氯通道阻滞剂二苯胺羧酸盐抑制。没有单层显示出葡萄糖转运的证据。
这三种未永生化的细胞系显示出隐窝样肠上皮细胞的形态、表型和转运特征。信使核糖核酸表达模式提示其具有促进生长和免疫调节作用。
