Human iPS cells engender corneal epithelial stem cells with holoclone-forming capabilities
Shinya Watanabe,
Ryuhei Hayashi,
Yuzuru Sasamoto,
Motokazu Tsujikawa,
Bruce R. Ksander,
Markus H. Frank,
Andrew J. Quantock,
Natasha Y. Frank,
Kohji Nishida
Affiliations
Shinya Watanabe
Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan
Ryuhei Hayashi
Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan; Department of Stem Cells and Applied Medicine, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan; Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives, Osaka University, Suita, Osaka 565-0871, Japan; Corresponding author
Yuzuru Sasamoto
Division of Genetics, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA
Motokazu Tsujikawa
Department of Biomedical Informatics, Osaka University Graduate School of Medicine, Division of Health Sciences, Suita, Osaka 565-0871, Japan
Bruce R. Ksander
Massachusetts Eye and Ear, Schepens Eye Research Institute, Harvard Medical School, Boston, MA 02114, USA
Markus H. Frank
Transplant Research Program, Division of Nephrology, Boston Children's Hospital, Harvard Medical School, Boston, MA 02115, USA; Harvard Stem Cell Institute, Harvard University, Boston, MA 02138, USA; School of Medical and Health Sciences, Edith Cowan University, Perth, WA 6027, Australia
Andrew J. Quantock
School of Optometry and Vision Sciences, Cardiff University, Cardiff CF24 4HQ, Wales, UK
Natasha Y. Frank
Division of Genetics, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA; Harvard Stem Cell Institute, Harvard University, Boston, MA 02138, USA; Department of Medicine, VA Boston Healthcare System, Boston, MA 02130, USA
Kohji Nishida
Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka 565-0871, Japan; Integrated Frontier Research for Medical Science Division, Institute for Open and Transdisciplinary Research Initiatives, Osaka University, Suita, Osaka 565-0871, Japan; Corresponding author
Summary: Human induced pluripotent stem cells (hiPSCs) can generate a multiplicity of organoids, yet no compelling evidence currently exists as to whether or not these contain tissue-specific, holoclone-forming stem cells. Here, we show that a subpopulation of cells in a hiPSC-derived corneal epithelial cell sheet is positive for ABCB5 (ATP-binding cassette, sub-family B, member 5), a functional marker of adult corneal epithelial stem cells. These cells possess remarkable holoclone-forming capabilities, which can be suppressed by an antibody-mediated ABCB5 blockade. The cell sheets are generated from ABCB5+ hiPSCs that first emerge in 2D eye-like organoids around six weeks of differentiation and display corneal epithelial immunostaining characteristics and gene expression patterns, including sustained expression of ABCB5. The findings highlight the translational potential of ABCB5-enriched, hiPSC-derived corneal epithelial cell sheets to recover vision in stem cell-deficient human eyes and represent the first report of holoclone-forming stem cells being directly identified in an hiPSC-derived organoid.
