Differentiation of a clone isolated from the HT29 cell line: polarized distribution of histocompatibility antigens (HLA) and of transferrin receptors.
The HT29 cell line, derived from a human colon adenocarcinoma, is able to differentiate if galactose replaces glucose in the culture medium. We have isolated a clone (HT29-18) from this cell line which displays differentiated properties of the parent cell line. HT29-18 cells grown in glucose-containing medium form multiple layers of round cells without specific cell-cell adhesion. In contrast, when grown in galactose-containing medium, they form a monolayer with tight junctions and exhibit a well differentiated brush border at their apical membrane, which faces the culture medium. The polarized properties of HT29-18 cells grown in galactose-containing medium were demonstrated by immunofluorescent techniques with antibodies against 2 plasma membrane proteins. Class I histocompatibility antigens (HLA) and transferrin receptors, 2 well characterized integral membrane proteins, are uniformly distributed on the cell surface of undifferentiated HT29-18 cells, but acquire a polarized distribution during differentiation, localized on the basolateral membranes and absent from the apical surface. Binding of 125I-labeled transferrin was used to determine transferrin receptor distribution on apical and basolateral membranes. Functional tight junctions in the differentiated cultures were demonstrated, as the monolayer was impermeable to a permeation dye (ruthenium red) as well as to antibodies. The sealing of these tight junctions is, as in vivo, Ca++-dependent as they could be opened by a short incubation in Ca++-free medium.
Godefroy
O
O
Département de Biologie Moléculaire, Institut Pasteur, Paris, France.
Huet
C
C
Blair
L A
LA
Sahuquillo-Merino
C
C
Louvard
D
D
D000230:Adenocarcinoma / Q000276:immunology / Q000648:ultrastructure; D002450:Cell Communication; D002455:Cell Division; D002460:Cell Line; D002462:Cell Membrane / Q000032:analysis / Q000648:ultrastructure; D002471:Cell Transformation, Neoplastic / Q000276:immunology / Q000648:ultrastructure; D003110:Colonic Neoplasms / Q000276:immunology / Q000648:ultrastructure; D003470:Culture Media; D005690:Galactose; D005947:Glucose; D006680:HLA Antigens / Q000032:analysis*; D006801:Humans; D008565:Membrane Proteins / Q000032:analysis; D011990:Receptors, Transferrin / Q000032:analysis*; D014168:Transferrin / Q000378:metabolism; D014407:Tumor Cells, Cultured / Q000276:immunology / Q000378:metabolism / Q000648:ultrastructure*
D003470:Culture Media; D006680:HLA Antigens; D008565:Membrane Proteins; D011990:Receptors, Transferrin; D014168:Transferrin; D005947:Glucose; D005690:Galactose