title: Usefulness of a novel Caco-2 cell perfusion system II. Characterization of monolayer properties and peptidase activity.
abstract: In this study, the enzymatic activity and the influence of support filters and extracellular matrix proteins on the differentiation of Caco-2 cells grown in a perfusion system (Minucells and MinutissueTM) were examined and compared to traditional culturing approaches. Differences were observed regarding the differentiation of Caco-2 cells using the traditional approach and perfusion system such that the cell monolayers grown in a perfusion system showed a significant increase in dipeptidase activities (18.20 +/- 0.43nmol x min(-1) x cm(-2)) compared to the cells cultivated using the 21-day protocol (9.45 +/- 0.50 nmol x min(-1) x cm(-2)). The peptidase activity of Caco-2 cells was strikingly inhibited when Matrigel extracellular protein was used for coating polycarbonate support filters. While the enzymatic activities of the cell monolayers differentiated in the perfusion system were up-regulated, the transepithelial electrical resistance values of the cell monolayers (171 +/- 52 and 251 +/- 62 omega x cm2 for polycarbonate and polyester, respectively) decreased compared to the traditional Snapwell inserts (644 +/- 119 omega x cm2). The results suggested that the perfusion systems were useful permeability models which reduce workload, resources and manpower needed to obtain useful Caco-2 monolayers. In addition, the approach offers an efficient tool for long-term culturing of highly differentiated Caco-2 cell monolayers.
authors.affiliation: Tibotec BVBA, Mechelen, Belgium.
authors.forename: C
authors.initials: C
authors.lastname: Masungi
authors.forename: J
authors.initials: J
authors.lastname: Mensch
authors.forename: B
authors.initials: B
authors.lastname: Willems
authors.forename: A
authors.initials: A
authors.lastname: Van Dijck
authors.forename: C
authors.initials: C
authors.lastname: Borremans
authors.forename: M
authors.initials: M
authors.lastname: Noppe
authors.forename: M E
authors.initials: ME
authors.lastname: Brewster
authors.forename: P
authors.initials: P
authors.lastname: Augustijns
mesh_terms: D000465:Algorithms; D000626:Aminopeptidases / Q000378:metabolism; D018938:Caco-2 Cells / Q000166:cytology* / Q000201:enzymology*; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D019070:Cell Lineage; D003470:Culture Media; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases / Q000378:metabolism; D017097:Electric Impedance; D006801:Humans; D018613:Microscopy, Confocal; D010447:Peptide Hydrolases / Q000378:metabolism*; D010477:Perfusion
chemical_list: D003470:Culture Media; D010447:Peptide Hydrolases; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases