April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Human Embryonic Stem Cell-derived Retinal Pigment Epithelial Cells Express Functional Multidrug Resistance Associated Protein (MRP) 1
Author Affiliations & Notes
  • Kati M. Juuti-Uusitalo
    Regea Institute for Regenerative Medicin, Regea Institute for Regenerative Medicine, Tampere, Finland
  • Hanna Vaajasaari
    Regea Institute for Regenerative Medicin, Regea Institute for Regenerative Medicine, Tampere, Finland
    Department of Ophthalmology, Institute of Clinical Medicine, University of Eastern Finland, Finland
  • Tuomas Ryhänen
    Department of Ophthalmology, Institute of Clinical Medicine, University of Eastern Finland, Finland
  • Tanja Ilmarinen
    Regea Institute for Regenerative Medicin, Regea Institute for Regenerative Medicine, Tampere, Finland
  • Susanna Narkilahti
    Regea Institute for Regenerative Medicin, Regea Institute for Regenerative Medicine, Tampere, Finland
  • Riitta Suuronen
    Regea Institute for Regenerative Medicin, Regea Institute for Regenerative Medicine, Tampere, Finland
  • Eliisa Mannermaa
    Department of Ophthalmology, Institute of Clinical Medicine, University of Eastern Finland, Finland
  • Kai Kaarniranta
    Department of Ophthalmology, Institute of Clinical Medicine, University of Eastern Finland, Finland
  • Heli Skottman
    Regea Institute for Regenerative Medicin, Regea Institute for Regenerative Medicine, Tampere, Finland
  • Footnotes
    Commercial Relationships  Kati M. Juuti-Uusitalo, None; Hanna Vaajasaari, None; Tuomas Ryhänen, None; Tanja Ilmarinen, None; Susanna Narkilahti, None; Riitta Suuronen, None; Eliisa Mannermaa, None; Kai Kaarniranta, None; Heli Skottman, None
  • Footnotes
    Support  Academy of Finland (218050), TEKES- the Finnish Funding Agency for Technology and Innovation (project StemInClin)
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 3188. doi:
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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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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: : 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.

Methods: : 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.

Results: : 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.

Conclusions: : 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.

Keywords: retinal pigment epithelium • retinal development • age-related macular degeneration 
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