Formation of multilayers in the Caco-2 cell culture model : A confocal laser scanning microscopy study

To introduce confocal laser scanning microscopy (CLSM) combined with digital image restoration to characterise Caco-2 cells under different culture conditions, and thus to define additional valid criteria for the optimisation of culture models. Growth curves were established and transepithelial elec...

Full description

Saved in:
Bibliographic Details
Published in:Pharmaceutical research 2000-04, Vol.17 (4), p.460-465
Main Authors: ROTHEN-RUTISHAUSER, B, BRAUN, A, GÜNTHERT, M, WUNDERLI-ALLENSPACH, H
Format: Article
Language:English
Subjects:
Adenocarcinoma - pathology
Amino acids
Analysis
Biological and medical sciences
Caco-2 Cells
Cell culture
Cell Division
Colonic Neoplasms - pathology
Cytoskeleton - ultrastructure
General pharmacology
Humans
Laboratories
Lasers
Medical sciences
Microscopy
Microscopy, Confocal - methods
Penicillin
Permeability
Pharmacology. Drug treatments
Sodium
Citations: Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:To introduce confocal laser scanning microscopy (CLSM) combined with digital image restoration to characterise Caco-2 cells under different culture conditions, and thus to define additional valid criteria for the optimisation of culture models. Growth curves were established and transepithelial electrical resistance (TEER) measured for cells grown in EMEM or DMEM medium on Cyclopore membranes. Cytoskeleton, cell nuclei and tight junctions (TJ) were investigated by CLSM. Cultures reached a plateau of approximately 4.5 x 10(5) cells/cm2 after approximately 10 days. At the same time TEER reached 750 omega cm2. An irregular, fairly complete network of TJ was present at confluence (approximately 2 d). Between 15 and 30 days a regular TJ network was established. Cells formed mixed mono- and multilayers under most conditions with two exceptions: flat monolayers were observed on polycarbonate filters with EMEM and with the Biocoat intestinal epithelium differentiation environment system. In multilayers TJ were found in the upper as well as in the lower cell layers although the regular vertical polarity was disturbed. CLSM represents an important tool to investigate the cytoarchitecture of Caco-2 cells. 3D-analysis of confocal data gives important clues on the characteristics of cell layers and thus helps to validate optimisation strategies.
ISSN:0724-8741
1573-904X
DOI:10.1023/A:1007585105753