June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Aquaporin expression and function in human pluripotent stem cell -derived retinal pigment epithelial cells
Author Affiliations & Notes
  • Kati Juuti-Uusitalo
    Institute of Biomedical Technology, University of Tampere, Tampere, Finland
  • Christine Delporte
    Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium
  • Francoise Gregoire
    Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium
  • Jason Perret
    Laboratory of Pathophysiological and Nutritional Biochemistry, Université Libre de Bruxelles, Brussels, Belgium
  • Virpi Savolainen
    Department of Biomedical Engineering, Tampere University of Technology, Tampere, Finland
  • Soile Nymark
    Department of Biomedical Engineering, Tampere University of Technology, Tampere, Finland
  • Jari Hyttinen
    Department of Biomedical Engineering, Tampere University of Technology, Tampere, Finland
  • Hannu Uusitalo
    SILK Department of Ophthalmology, University of Tampere, Tampere, Finland
    Eye Center, Tampere University Hospital, Tampere, Finland
  • Francois Willermain
    Department of Ophthalmology, CHU St Pierre and Brugmann and I.R.I.B.H.M, Brussels, Belgium
  • Heli Skottman
    Institute of Biomedical Technology, University of Tampere, Tampere, Finland
  • Footnotes
    Commercial Relationships Kati Juuti-Uusitalo, None; Christine Delporte, None; Francoise Gregoire, None; Jason Perret, None; Virpi Savolainen, None; Soile Nymark, None; Jari Hyttinen, None; Hannu Uusitalo, None; Francois Willermain, None; Heli Skottman, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2238. doi:
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      Kati Juuti-Uusitalo, Christine Delporte, Francoise Gregoire, Jason Perret, Virpi Savolainen, Soile Nymark, Jari Hyttinen, Hannu Uusitalo, Francois Willermain, Heli Skottman; Aquaporin expression and function in human pluripotent stem cell -derived retinal pigment epithelial cells. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2238.

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Abstract

Purpose: The expression of aquaporin water channel genes are shown to be affected in several pathological conditions of retina, such as in diabetic retinopathy, retinal ischemia and in autoimmune uveitis. Human native retinal pigment epithelial (RPE) cells and immortalized human RPEs are formerly shown to express aquaporins, still the expression of aquaporins in stem cell derived RPE have not been previously elucidated. The objective of this study was to determine the expression of several aquaporin genes (aquaporin1,- 3, -4, -5, -6, -7, -10, -11 and -12) and assess the localization of aquaporin 1 water channel protein in human embryonic (hESCs) and induced pluripotent stem cells (hiPSCs) derived RPE cells.

Methods: hESC- and hiPSC derived RPE cells were grown as monolayer in serum-free media. The expression of aquaporin genes was determined with qRT-PCR. The localization of AQP1-protein was studied with confocal microscopy. Finally, the functionality of aquaporins was assessed with dye accumulation test in presence and absence of AQP inhibitors in hypotonic and isotonic solutions.

Results: The aquaporin1,-3, -4, -5, -6, -7, -10, and -11 genes were shown to be expressed in hESC- and hiPSC derived RPE cells. Aquaporin1 water channel gene expression was significantly upregulated during RPE cell differentiation. The immunofluorescence investigation revealed that in polarized RPE cells the localization of aquaporin1 was more pronounced, especially on apical cell membrane. Lastly, the aquaporin inhibitors significantly reduced the dye accumulation to cells that were bathed in hypotonic solution.

Conclusions: For the first time we have shown that aquaporin water channel genes are expressed by hESC- and hiPSC derived RPE cells. Moreover, the localization of aquaporin1 on plasma membrane during the maturation suggests that the aquaporin1 protein is functional in mature hESC-derived RPE cells. According to this data hESC- and hiPSC derived RPE cells, cultured and differentiated under serum-free conditions resemble their native counterpart in the human body.

Keywords: 701 retinal pigment epithelium • 721 stem cells • 497 development  
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