title: Filter-cultured ARPE-19 cells as outer blood-retinal barrier model.
abstract: Retinal pigment epithelium (RPE) regulates drug transfer between posterior eye segment and blood circulation, but there is no established RPE cell model for drug delivery studies. We evaluated ARPE-19 filter culture model for this purpose. Passive permeability of 6-carboxyfluorescein, betaxolol and FITC-dextran (40kDa) and active transport of 6-carboxyfluorescein, sodium fluorescein, rhodamine 123, cyclosporine A and digoxin in ARPE-19 model were investigated and compared with isolated bovine RPE-choroid tissue. In addition, barrier properties, and mRNA expression of RPE-specific and melanogenesis-related genes (RPE65, VMD2, CRALBP, OTX-2, MITF-A, TRP-1, tyrosinase) were measured in various culture conditions. The filter grown ARPE-19 cell model showed reasonable barrier properties (TER close to 100Omegacm(2)), but its permeability was slightly higher than that of isolated bovine RPE/choroid specimens. In active transport studies the ARPE-19 model mimics qualitatively the permeability profile of bovine RPE-choroid, but ARPE-19 model underestimates the importance of active transport relative to passive diffusion. Long-term filter-cultured ARPE-19 cells expressed various RPE-specific and melanogenesis-related genes at higher levels than the ARPE-19 cells cultured short-term in flasks. ARPE-19 model can be used to study drug permeation processes in the RPE.
authors.affiliation: Department of Pharmaceutics, University of Kuopio, Yliopistonranta 1 C, 70210 Kuopio, Finland.
authors.forename: Eliisa
authors.initials: E
authors.lastname: Mannermaa
authors.forename: Mika
authors.initials: M
authors.lastname: Reinisalo
authors.forename: Veli-Pekka
authors.initials: VP
authors.lastname: Ranta
authors.forename: Kati-Sisko
authors.initials: KS
authors.lastname: Vellonen
authors.forename: Heidi
authors.initials: H
authors.lastname: Kokki
authors.forename: Anni
authors.initials: A
authors.lastname: Saarikko
authors.forename: Kai
authors.initials: K
authors.lastname: Kaarniranta
authors.forename: Arto
authors.initials: A
authors.lastname: Urtti
mesh_terms: D001693:Biological Transport, Active; D001813:Blood-Retinal Barrier / Q000378:metabolism*; D002460:Cell Line; D002463:Cell Membrane Permeability; D016764:Cell Polarity; D015194:Diffusion Chambers, Culture; D005136:Eye Proteins / Q000235:genetics / Q000378:metabolism; D015870:Gene Expression; D006801:Humans; D057927:Hydrophobic and Hydrophilic Interactions; D008543:Melanins / Q000096:biosynthesis; D008970:Molecular Weight; D010599:Pharmacokinetics; D012333:RNA, Messenger / Q000378:metabolism; D055213:Retinal Pigment Epithelium / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
chemical_list: D005136:Eye Proteins; D008543:Melanins; D012333:RNA, Messenger