Xeno-Free and Defined Human Embryonic Stem Cell-Derived Retinal Pigment Epithelial Cells Functionally Integrate in a Large-Eyed Preclinical Model

Stem Cell Reports. 2016 Jan 12;6(1):9-17. doi: 10.1016/j.stemcr.2015.11.008. Epub 2015 Dec 24.


Human embryonic stem cell (hESC)-derived retinal pigment epithelial (RPE) cells could replace lost tissue in geographic atrophy (GA) but efficacy has yet to be demonstrated in a large-eyed model. Also, production of hESC-RPE has not yet been achieved in a xeno-free and defined manner, which is critical for clinical compliance and reduced immunogenicity. Here we describe an effective differentiation methodology using human laminin-521 matrix with xeno-free and defined medium. Differentiated cells exhibited characteristics of native RPE including morphology, pigmentation, marker expression, monolayer integrity, and polarization together with phagocytic activity. Furthermore, we established a large-eyed GA model that allowed in vivo imaging of hESC-RPE and host retina. Cells transplanted in suspension showed long-term integration and formed polarized monolayers exhibiting phagocytic and photoreceptor rescue capacity. We have developed a xeno-free and defined hESC-RPE differentiation method and present evidence of functional integration of clinically compliant hESC-RPE in a large-eyed disease model.

Keywords: cellular therapy; differentiation; human embryonic stem cells; laminin; optical coherence tomography; rabbit; retinal pigment epithelium; transplantation; xeno-free.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Culture Techniques
  • Cell Differentiation / drug effects
  • Cell Differentiation / physiology*
  • Cell Line
  • Culture Media / chemistry
  • Culture Media / pharmacology
  • Disease Models, Animal
  • Geographic Atrophy / physiopathology*
  • Geographic Atrophy / therapy
  • Human Embryonic Stem Cells / cytology
  • Human Embryonic Stem Cells / physiology*
  • Human Embryonic Stem Cells / transplantation
  • Humans
  • Laminin / metabolism
  • Microscopy, Confocal
  • Rabbits
  • Retinal Pigment Epithelium / cytology
  • Retinal Pigment Epithelium / physiology*
  • Stem Cell Transplantation / methods
  • Time-Lapse Imaging
  • Transplantation, Heterologous
  • Xenobiotics / chemistry
  • Xenobiotics / pharmacology


  • Culture Media
  • Laminin
  • Xenobiotics
  • laminin 11 protein, human