Filter-cultured ARPE-19 cells as outer blood-retinal barrier model.
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.
Mannermaa
Eliisa
E
Department of Pharmaceutics, University of Kuopio, Yliopistonranta 1 C, 70210 Kuopio, Finland.
Reinisalo
Mika
M
Ranta
Veli-Pekka
VP
Vellonen
Kati-Sisko
KS
Kokki
Heidi
H
Saarikko
Anni
A
Kaarniranta
Kai
K
Urtti
Arto
A
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
D005136:Eye Proteins; D008543:Melanins; D012333:RNA, Messenger