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Kati M. Juuti-Uusitalo, Hanna Vaajasaari, Tuomas Ryhänen, Tanja Ilmarinen, Susanna Narkilahti, Riitta Suuronen, Eliisa Mannermaa, Kai Kaarniranta, Heli Skottman; Human Embryonic Stem Cell-derived Retinal Pigment Epithelial Cells Express Functional Multidrug Resistance Associated Protein (MRP) 1. Invest. Ophthalmol. Vis. Sci. 2011;52(14):3188.
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The objective of this study was to determine the expression and functionality of multidrug-resistance -associated protein (MRP) both in immature and mature human embryonic stem cell (hESC) -derived retinal pigment epithelium (RPE) cells which have been cultured under serum-free conditions.
For all experiments hESC-derived RPE cells were cultured as monolayer in serum-free media. Western blotting was employed to examine the presence of MRP1 protein. The localization of MRP-protein was studied with confocal microscopy. Finally, calcein-AM uptake was performed to assess MRP1 activity. The inhibitors, verapamil, progesterone, cyclosporine and MK571, were used to block the transport activity.
MRP1 was expressed both in immature and mature hESC-derived RPE cells. The immunofluorescence investigation revealed that in mature and polarized RPE cells the localization was more pronounced, especially on apical cell membrane. Finally, the MRP1 expressed in human hESC-derived RPE cells was active.
For the first time we have shown that hESC-derived RPE cells express functional MRP1 efflux pump. This suggests that hESC-derived RPE cells, cultured and differentiated under serum-free conditions showing typical molecular and functional characteristics of RPE cells, resemble their native counterpart in the human body.
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