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A new efficient protocol for directed differentiation of retinal pigmented epithelial cells from normal and retinal disease induced pluripotent stem cells.

Abstract
We describe a new, efficient protocol that involves the serial addition of noggin, basic fibroblast growth factor (bFGF), retinoic acid, and sonic hedgehog (Shh) for the differentiation of human induced pluripotent stem cells (hiPSC) to retinal pigmented epithelium (RPE) in a serum- and feeder-free adherent condition. hiPSC-RPE cells exhibited RPE morphology and specific molecular markers. Additionally, several hiPSC lines were generated from retinal-specific patients with Leber's congenital amaurosis, Usher syndrome, two patients with retinitis pigmentosa, and a patient with Leber's hereditary optic neuropathy. The RPE cells generated from these disease-specific hiPSCs expressed specific markers by the same RPE lineage-directed differentiation protocol. These findings indicate a new short-term, simple, and efficient protocol for differentiation of hiPSCs to RPE cells. Such specific retinal disease-specific hiPSCs offer an unprecedented opportunity to recapitulate normal and pathologic formation of human retinal cells in vitro, thereby enabling pharmaceutical screening, and potentially autologous cell replacement therapies for retinal diseases.
AuthorsAzadeh Zahabi, Ebrahim Shahbazi, Hamid Ahmadieh, Seyedeh-Nafiseh Hassani, Mehdi Totonchi, Adeleh Taei, Najmehsadat Masoudi, Marzieh Ebrahimi, Nasser Aghdami, Ali Seifinejad, Faramarz Mehrnejad, Narsis Daftarian, Ghasem Hosseini Salekdeh, Hossein Baharvand
JournalStem cells and development (Stem Cells Dev) Vol. 21 Issue 12 Pg. 2262-72 (Aug 10 2012) ISSN: 1557-8534 [Electronic] United States
PMID22145677 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
Chemical References
  • Antigens, Differentiation
Topics
  • Adolescent
  • Adult
  • Antigens, Differentiation (genetics, metabolism)
  • Cell Culture Techniques
  • Cell Differentiation
  • Cell Shape
  • Cells, Cultured
  • Child
  • Female
  • Gene Expression
  • Humans
  • Induced Pluripotent Stem Cells (metabolism, physiology)
  • Karyotype
  • Male
  • Middle Aged
  • Phenotype
  • Real-Time Polymerase Chain Reaction
  • Retinal Diseases (pathology)
  • Retinal Pigment Epithelium (metabolism, pathology)

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