September 2016
Volume 57, Issue 12
Open Access
ARVO Annual Meeting Abstract  |   September 2016
Generation of human retinal progenitor cells from human induced pluripotent stem cells-derived spherical neural masses and their survival and differentiation in a mouse retina
Author Affiliations & Notes
  • Cheolmin Yun
    Ophthalmology, Korea University College of Medicine, Seoul, Korea (the Republic of)
  • Jaeryung Oh
    Ophthalmology, Korea University College of Medicine, Seoul, Korea (the Republic of)
  • Boram Lee
    Ophthalmology, Korea University College of Medicine, Seoul, Korea (the Republic of)
  • Seong-Woo Kim
    Ophthalmology, Korea University College of Medicine, Seoul, Korea (the Republic of)
  • Dongho Geum
    Biomedical Sciences, Korea University College of Medicine, Seoul, Korea (the Republic of)
  • Kuhl Huh
    Ophthalmology, Korea University College of Medicine, Seoul, Korea (the Republic of)
  • Footnotes
    Commercial Relationships   Cheolmin Yun, None; Jaeryung Oh, None; Boram Lee, None; Seong-Woo Kim, None; Dongho Geum, None; Kuhl Huh, None
  • Footnotes
    Support  None
Investigative Ophthalmology & Visual Science September 2016, Vol.57, 6065. doi:
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      Cheolmin Yun, Jaeryung Oh, Boram Lee, Seong-Woo Kim, Dongho Geum, Kuhl Huh; Generation of human retinal progenitor cells from human induced pluripotent stem cells-derived spherical neural masses and their survival and differentiation in a mouse retina. Invest. Ophthalmol. Vis. Sci. 2016;57(12):6065.

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

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Abstract

Purpose : To differentiate neural retinal progenitor cells (RPCs) from human induced pluripotent stem cells (hiPSCs) - derived spherical neural masses (SNMs) and investigate whether the RPCs survive and differentiate in a mouse retina.

Methods : SNMs were derived from hiPSCs by going through embryoid body formation, neural precursors selection, neural precursors expansion and dissection of neural structures. RPCs were differentiated from SNMs with noggin/fibroblast growth factor-basic (bFGF)/Dickkopf-1(Dkk-1)/Insulin-like growth factor 1 (IGF-1)/fibroblast growth factor 9 (FGF-9) protocol for three weeks. Differentiation profiles of human RPCs in vitro were analyzed by immunocytochemistry and quantitative reverse transcription polymerase chain reaction (RT-PCR) analysis. Human RPCs were transplanted into the mouse (adult 8-12 weeks old C57BL/6) retina. Transplanted eye was examined post-operatively at three months and analyzed with immunohistochemistry.

Results : Human RPCs from hiPSCs - derived SNMs expressed eye field markers (Paired box 6, PAX6) and early neural retinal markers (Ceh-10 homeodomain containing homolog, CHX10), except for photoreceptor markers (S-cone opsin [OPN1SW]). RT-PCR revealed that early neural retinal markers (achaete-scute complex homolog 1 [MASH], mouse atonal homolog 5 [MATH5], neurogenic differentiation 1 [NEUROD1]) and retinal fate markers (brain-specific homeobox/POU domain transcription factor 3B [BRN3B], recoverin) were up-regulated, while the marker of retinal pigment epithelium (microphthalmia-associated transcription factor [MITF]) only showed a slight upregulation. The transplanted cells in the retina of mouse were matured to express markers mature retinal cells (OPN1SW) and human nuclei (HNu) on immunohistochemistry at three months after transplantation.

Conclusions : Human RPCs with neural retinal fate were differentiated from the hiPSCs-derived SNMs with noggin/bFGF/Dkk-1/IGF-1/FGF-9 protocol in vitro. Intraocular transplanted RPCs survived and differentiated into retinal photoreceptor cells in a mouse retina. Development of RPCs using SNMs may be a fast and useful method for differentiation of neural retinal cells.

This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.

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