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Open Access Usefulness of a novel Caco-2 cell perfusion system II. Characterization of monolayer properties and peptidase activity

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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 ± 0.43 nmol · min1 · cm–2) compared to the cells cultivated using the 21-day protocol (9.45 ± 0.50 nmol · min–1 · 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 Ω · cm2 for polycarbonate and polyester, respectively) decreased compared to the traditional Snapwell inserts (644 ± 119 Ω · 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.

Document Type: Research Article

DOI: https://doi.org/10.1691/ph.2009.8592

Affiliations: 1: Tibotec BVBA, Mechelen, Belgium 2: Pharmaceutical Sciences, Johnson & Johnson Pharmaceutical Research and Development, a Division of Janssen Pharmaceutica, Beerse, Belgium 3: Drug Metabolism and Pharmacokinetics, Johnson & Johnson Pharmaceutical Research and Development, a Division of Janssen Pharmaceutica, Beerse, Belgium 4: Clinical Supply Unit, Johnson & Johnson Pharmaceutical Research and Development, a Division of Janssen Pharmaceutica, Beerse, Belgium 5: IM/CNS Research and Early Development, Johnson & Johnson Pharmaceutical Research and Development, a Division of Janssen Pharmaceutica, Beerse, Belgium 6: Pharmaceutical Sciences, Johnson & Johnson Pharmaceutical Research and Development, a Division of Janssen Pharmaceutica, Beerse, 2340, Belgium, Email: mbrewste@prdbe.jnj.com 7: Laboratory for Pharmacotechnology and Biopharmacy, Katholieke Universiteit Leuven, O&N, Gasthuisberg, Leuven, Belgium

Publication date: 2009-01-01

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  • Pharmazie is a leading journal in the field of pharmaceutical sciences. As a peer-reviewed scientific journal, Pharmazie is regularly indexed in the relevant databases like Web of science, Journal Citation Reports and many others. The journal is open for submissions from the whole spectrum of pharnaceutical sciences including Pharmaceutical Chemistry, Experimental and Clinical Pharmacology, Drug Analysis, Pharmaceutics, Pharmaceutical Biology, Clinical Pharmacy etc.
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