Immunologic, functional, and morphological characterization of three new human small intestinal epithelial cell lines.
OBJECTIVE
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.


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Pang
G
G
Faculty of Medicine and Australian Institute of Mucosal Immunology, University of Newcastle, New South Wales, Australia.
Buret
A
A
O'Loughlin
E
E
Smith
A
A
Batey
R
R
Clancy
R
R
D000368:Aged; D001483:Base Sequence; D002460:Cell Line; D018118:Chloride Channels / Q000187:drug effects / Q000378:metabolism; D016207:Cytokines / Q000235:genetics; D004186:Disaccharidases / Q000378:metabolism; D004386:Duodenum / Q000166:cytology* / Q000276:immunology / Q000502:physiology; D004847:Epithelial Cells; D004848:Epithelium / Q000276:immunology / Q000502:physiology; D005260:Female; D015870:Gene Expression; D006801:Humans; D015151:Immunoblotting; D007150:Immunohistochemistry; D017136:Ion Transport; D007633:Keratins / Q000378:metabolism; D008297:Male; D008875:Middle Aged; D008969:Molecular Sequence Data; D009077:Mucins / Q000378:metabolism; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000378:metabolism
D018118:Chloride Channels; D016207:Cytokines; D009077:Mucins; D012333:RNA, Messenger; D007633:Keratins; D004186:Disaccharidases