Mucin production by colon cancer cells cultured in serum-free medium.
Although many colon cancer cell lines are available for study, few of them exhibit differentiated properties. When cultured in medium containing fetal bovine serum, WiDr cells (WiDr-FBS) show an undifferentiated phenotype: growth as a multilayer of cells adherent to plastic and lack of polarization, brush border, and mucin vacuoles. In contrast, WiDr cells cultured in a chemically-defined serum-free medium containing insulin, transferrin and selenium (WiDr-ITS) grow as clusters of nonadherent cells with abundant desmosomes and tight junctions, microvilli and electron-lucid vacuoles. As WiDr-FBS cells, WiDr-ITS are not polarized. WiDr-ITS cells show a marked enhancement in mucin synthesis as demonstrated by: periodic acid-Schiff and Alcian blue stains, electron microscopy, immunohistochemistry using monoclonal antibodies (MAbs) reactive with mucin-associated epitopes, immune electron microscopy and immunochemical analysis using Western blots. In comparison with WiDr-FBS cells, WiDr-ITS cells showed strong expression of Tn, sialyl-Tn, blood group A and CEA. When mouse MAbs were used, higher levels of the MUCI gene product were detected in WiDr-ITS than in WiDr-FBS cells. The full spectrum of phenotypic changes was observed after I month of culture in ITS medium, and transfer of WiDr-ITS cells to FBS medium was accompanied by a partial phenotypic reversal, suggesting that these phenotypic changes result from an adaptative--rather than selective--process.
Real
F X
FX
Departament d'Immunologia, Institut Municipal d'Investigació Mèdica, Barcelona, Spain.
Egea
G
G
Francí
C
C
Schüssler
M H
MH
Xu
M
M
Welt
S
S
D000423:Alcian Blue; D000911:Antibodies, Monoclonal; D000941:Antigens / Q000032:analysis / Q000276:immunology; D001789:Blood Group Antigens / Q000276:immunology; D015153:Blotting, Western; D002455:Cell Division; D003110:Colonic Neoplasms / Q000378:metabolism* / Q000648:ultrastructure; D003470:Culture Media; D006651:Histocytochemistry; D007150:Immunohistochemistry; D008854:Microscopy, Electron; D016253:Microscopy, Immunoelectron; D009077:Mucins / Q000096:biosynthesis* / Q000276:immunology; D010503:Periodic Acid-Schiff Reaction; D010641:Phenotype; D013194:Staining and Labeling; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D000941:Antigens; D001789:Blood Group Antigens; D003470:Culture Media; D009077:Mucins; D000423:Alcian Blue