In vitro differentiation of retinal cells from human pluripotent stem cells by small-molecule induction

The use of stem-cell therapy to treat retinal degeneration holds great promise. However, definitive methods of retinal differentiation that do not depend on recombinant proteins produced in animal or Escherichia coli cells have not been devised. Here, we report a defined culture method using low-mol...

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Veröffentlicht in:Journal of cell science 2009-09, Vol.122 (17), p.3169-3179
Hauptverfasser: Osakada, Fumitaka, Jin, Zi-Bing, Hirami, Yasuhiko, Ikeda, Hanako, Danjyo, Teruko, Watanabe, Kiichi, Sasai, Yoshiki, Takahashi, Masayo
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container_end_page 3179
container_issue 17
container_start_page 3169
container_title Journal of cell science
container_volume 122
creator Osakada, Fumitaka
Jin, Zi-Bing
Hirami, Yasuhiko
Ikeda, Hanako
Danjyo, Teruko
Watanabe, Kiichi
Sasai, Yoshiki
Takahashi, Masayo
description The use of stem-cell therapy to treat retinal degeneration holds great promise. However, definitive methods of retinal differentiation that do not depend on recombinant proteins produced in animal or Escherichia coli cells have not been devised. Here, we report a defined culture method using low-molecular-mass compounds that induce differentiation of human embryonic stem (ES) cells and induced pluripotent stem (iPS) cells into retinal progenitors, retinal pigment epithelium cells and photoreceptors. The casein kinase I inhibitor CKI-7, the ALK4 inhibitor SB-431542 and the Rho-associated kinase inhibitor Y-27632 in serum-free and feeder-free floating aggregate culture induce retinal progenitors positive for RX, MITF, PAX6 and CHX10. The treatment induces hexagonal pigmented cells that express RPE65 and CRALBP, form ZO1-positive tight junctions and exhibit phagocytic functions. Subsequent treatment with retinoic acid and taurine induces photoreceptors that express recoverin, rhodopsin and genes involved in phototransduction. Both three-factor (OCT3/4, SOX2 and KLF4) and four-factor (OCT3/4, SOX2, KLF4 and MYC) human iPS cells could be successfully differentiated into retinal cells by small-molecule induction. This method provides a solution to the problem of cross-species antigenic contamination in cell-replacement therapy, and is also useful for in vitro modeling of development, disease and drug screening.
doi_str_mv 10.1242/jcs.050393
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subjects Amides - pharmacology
Animals
Benzamides - pharmacology
Cell Culture Techniques - methods
Cell Differentiation - drug effects
Cells, Cultured
Dioxoles - pharmacology
Gene Expression - drug effects
Humans
Isoquinolines - pharmacology
Mice
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - drug effects
Pluripotent Stem Cells - metabolism
Protein Kinase Inhibitors - pharmacology
Pyridines - pharmacology
Retina - cytology
Retina - drug effects
Retina - metabolism
title In vitro differentiation of retinal cells from human pluripotent stem cells by small-molecule induction
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