Usefulness of a novel Caco-2 cell perfusion system II. Characterization of monolayer properties and peptidase activity.
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.
Masungi
C
C
Tibotec BVBA, Mechelen, Belgium.
Mensch
J
J
Willems
B
B
Van Dijck
A
A
Borremans
C
C
Noppe
M
M
Brewster
M E
ME
Augustijns
P
P
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
D003470:Culture Media; D010447:Peptide Hydrolases; D000626:Aminopeptidases; D004152:Dipeptidyl-Peptidases and Tripeptidyl-Peptidases