June 2015
Volume 56, Issue 7
Free
ARVO Annual Meeting Abstract  |   June 2015
Xeno-free generation of human embryonic stem cells into retinal pigment epithelial cells and their integration as a polarized monolayer in a preclinical model
Author Affiliations & Notes
  • Sandra Petrus-Reurer
    Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden
    Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
  • Sonya Stenfelt
    Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
  • Alvaro Plaza-Reyes
    Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
  • Hammurabi Bartuma
    Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden
    Sankt Eriks Eye Hospital, Stockholm, Sweden
  • Liselotte Antonsson
    Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
  • Sarita Pauliina Panula
    Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
  • Helder Andre
    Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden
    Sankt Eriks Eye Hospital, Stockholm, Sweden
  • Fredrik Lanner
    Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
  • Outi Hovatta
    Clinical Sciences, Intervention and Technology, Karolinska Institutet, Stockholm, Sweden
  • Anders P Kvanta
    Clinical Neuroscience, Karolinska Institutet, Stockholm, Sweden
    Sankt Eriks Eye Hospital, Stockholm, Sweden
  • Footnotes
    Commercial Relationships Sandra Petrus-Reurer, None; Sonya Stenfelt, None; Alvaro Plaza-Reyes, None; Hammurabi Bartuma, None; Liselotte Antonsson, None; Sarita Pauliina Panula, None; Helder Andre, None; Fredrik Lanner, None; Outi Hovatta, None; Anders Kvanta, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2015, Vol.56, 4214. doi:
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      Sandra Petrus-Reurer, Sonya Stenfelt, Alvaro Plaza-Reyes, Hammurabi Bartuma, Liselotte Antonsson, Sarita Pauliina Panula, Helder Andre, Fredrik Lanner, Outi Hovatta, Anders P Kvanta; Xeno-free generation of human embryonic stem cells into retinal pigment epithelial cells and their integration as a polarized monolayer in a preclinical model. Invest. Ophthalmol. Vis. Sci. 2015;56(7 ):4214.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Purpose: Transplantation of human embryonic stem cell (hESC) derived retinal pigment epithelial (RPE) cells could be used to replace the tissue lost in the degenerative form of macular degeneration. The production of functional RPE cells have however not been generated in a xeno-free and defined manner which is critical for clinical compliance and to reduce immunogenicity.

Methods: Building on our recently developed methodology for xeno-free derivation of hESC, we now present in vitro differentiation into RPE cells by using a human recombinant laminin-based matrix and a xeno-free, chemically defined medium. Briefly, hESC are differentiated as embryoid bodies in suspension. Optic vesicle-like structures containing pigmented cells emerge following three weeks of differentiation and are manually isolated two weeks later. Enzymatically dissociated single cells are subsequently cultured in 2D cultures on laminin. Suspensions of differentiated cells were transplanted into albino rabbits and cell fate was analyzed by real-time high-resolution imaging and immunohistochemistry.

Results: This approach yields highly homogeneous populations of mature hESC-derived RPE cells that exhibit characteristics of native RPE cells such as morphology, pigmentation, expression and polarization of specific markers and phagocytic activity. Furthermore, upon subretinal transplantation into immunosuppressed albino rabbits, these cells form an organized, polarized and pigmented epithelial layer with maintained expression of RPE markers.

Conclusions: ‚ÄčAltogether, these findings prove that our xeno-free and chemically defined differentiation method could serve as a potential source of clinically compliant RPE cells for the development of a safe and efficient cell replacement therapy for age-related macular degeneration.

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