HCE-T cell-based permeability model: A well-maintained or a highly variable barrier phenotype?
The most extensively characterized human-derived cell line used in transcorneal permeability studies, in terms of passive transcellular and paracellular transport, transporter expression and metabolic enzymes, is the immortalized human corneal epithelial cell line (HCE-T). The purpose of this study is to describe the changes in the HCE-T barrier phenotype in vitro when valid cultivation conditions, in accordance with the standardized HCE-T cell-based model protocol, were employed. Evaluation of the structural and functional barrier properties revealed two different HCE-T barrier phenotypes, depending on the polycarbonate membrane pore size. Model I (pore size 0.4μm) was characterized by a multilayered HCE-T epithelium at the apical side and a weak barrier function (70-115Ω×cm2), whereas Model II (pore size 3μm) consisted of an apical lipophilic HCE-T monolayer and a basolateral lipophilic monolayer of migrated HCE-T cells that showed improved barrier properties (1700-2600Ω×cm2) compared with Model I. Considering the permeation of ophthalmic compounds and in vitro/ex vivo correlation, Model II was better able to predict transcorneal drug permeation. This study highlights the important aspects of HCE-T barrier phenotype variability that should be continuously monitored in the routine application of HCE-T cell-based models across both academic and pharmaceutical industry research laboratories.
University of Zagreb, Faculty of Pharmacy and Biochemistry, Department of Pharmaceutical Technology, A. Kovačića 1, 10000 Zagreb, Croatia.
Marina
M
Juretić
University of Zagreb, Faculty of Pharmacy and Biochemistry, Department of Pharmaceutical Technology, A. Kovačića 1, 10000 Zagreb, Croatia.
Bisera
B
Jurišić Dukovski
R&D, PLIVA Croatia Ltd, TEVA Group Member, Prilaz baruna Filipovića 25, 10000 Zagreb, Croatia.
Iva
I
Krtalić
Institut für Pharmazeutische Technologie, Technische Universität Braunschweig, Braunschweig, Germany.
Stephan
S
Reichl
R&D, PLIVA Croatia Ltd, TEVA Group Member, Prilaz baruna Filipovića 25, 10000 Zagreb, Croatia.
Biserka
B
Cetina-Čižmek
University of Zagreb, Faculty of Pharmacy and Biochemistry, Department of Pharmaceutical Technology, A. Kovačića 1, 10000 Zagreb, Croatia.
Jelena
J
Filipović-Grčić
University of Zagreb, Faculty of Pharmacy and Biochemistry, Department of Pharmaceutical Technology, A. Kovačića 1, 10000 Zagreb, Croatia. Electronic address: jlovric@pharma.hr.
Jasmina
J
Lovrić
University of Zagreb, Faculty of Pharmacy and Biochemistry, Department of Pharmaceutical Technology, A. Kovačića 1, 10000 Zagreb, Croatia. Electronic address: ipepic@pharma.hr.
Ivan
I
Pepić
D000818:Animals; D002461:Cell Line, Transformed; D002463:Cell Membrane Permeability; D019573:Epithelium, Corneal / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D008954:Models, Biological; D013552:Swine
Barrier properties; Cornea; Drug permeability; In vitro/ex vivo correlation; Ocular delivery