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Cell Stem Cell. 2015 Jul 2;17(1):101-15. doi: 10.1016/j.stem.2015.05.015.

Quantification of Retinogenesis in 3D Cultures Reveals Epigenetic Memory and Higher Efficiency in iPSCs Derived from Rod Photoreceptors.

Author information

1
Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
2
Department of Computational Biology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
3
Department of Cell Biology, The Scripps Research Institute, La Jolla, CA 92037, USA.
4
Mouse Genetics Core Facility, The Scripps Research Institute, La Jolla, CA 92037, USA.
5
Division of Basic Sciences, Fred Hutchinson Cancer Research Center, Seattle, WA 98108, USA.
6
Department of Ophthalmology, University of Tennessee Health Science Center, Memphis, TN 38163, USA.
7
Cell and Tissue Imaging Shared Resource, St. Jude Children's Research Hospital, Memphis, TN 38105, USA.
8
Department of Developmental Neurobiology, St. Jude Children's Research Hospital, Memphis, TN 38105, USA; Department of Ophthalmology, University of Tennessee Health Science Center, Memphis, TN 38163, USA; Howard Hughes Medical Institute, Chevy Chase, MD 20815, USA. Electronic address: michael.dyer@stjude.org.

Abstract

Cell-based therapies to treat retinal degeneration are now being tested in clinical trials. However, it is not known whether the source of stem cells is important for the production of differentiated cells suitable for transplantation. To test this, we generated induced pluripotent stem cells (iPSCs) from murine rod photoreceptors (r-iPSCs) and scored their ability to make retinae by using a standardized quantitative protocol called STEM-RET. We discovered that r-iPSCs more efficiently produced differentiated retinae than did embryonic stem cells (ESCs) or fibroblast-derived iPSCs (f-iPSCs). Retinae derived from f-iPSCs had fewer amacrine cells and other inner nuclear layer cells. Integrated epigenetic analysis showed that DNA methylation contributes to the defects in f-iPSC retinogenesis and that rod-specific CTCF insulator protein-binding sites may promote r-iPSC retinogenesis. Together, our data suggest that the source of stem cells is important for producing retinal neurons in three-dimensional (3D) organ cultures.

PMID:
26140606
PMCID:
PMC4547539
DOI:
10.1016/j.stem.2015.05.015
[Indexed for MEDLINE]
Free PMC Article

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