CD200 facilitates the isolation of corneal epithelial cells derived from human pluripotent stem cells
The in vitro induction of corneal epithelial cells (CECs) from human induced pluripotent stem cells (iPSCs) represents a new strategy for obtaining CE stem/progenitor cells for the surgical reconstruction of a diseased or injured ocular surface. The clinical promise of this strategy is considerable,...
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Published in | Scientific reports Vol. 8; no. 1; pp. 16550 - 11 |
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Main Authors | , , , , |
Format | Journal Article |
Language | English |
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08.11.2018
Nature Publishing Group UK |
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Abstract | The in vitro induction of corneal epithelial cells (CECs) from human induced pluripotent stem cells (iPSCs) represents a new strategy for obtaining CE stem/progenitor cells for the surgical reconstruction of a diseased or injured ocular surface. The clinical promise of this strategy is considerable, but if the approaches' potential is to be realised, robust methods for the purification of iPSC-derived CE lineage cells need to be developed to avoid contamination with other cells that may carry the risk of unwanted side effects, such as tumorigenesis. Experiments conducted here revealed that during CEC isolation, CD200-negative selection using a cell sorter considerably reduced the contamination of the cell population with various non-CECs compared with what could be achieved using TRA-1-60, a conventional negative marker for CECs. Furthermore, CD200-negative sorting did not affect the yield of CECs nor that of their stem/progenitor cells. Single-cell gene expression analysis for CEC sheets obtained using CD200-negative sorting showed that all analysed cells were CE-lineage cells, expressing PAX6, delta-N p63, and E-cadherin. Non-CECs, on the other hand, expressed non-CEC genes such as FGFR1 and RPE65. CD200, thus, represents a robust negative marker for purification of induced CE lineage cells, which is expressed by undifferentiated iPSCs and non-CECs, including iPSC-derived neural and retinal cells. |
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AbstractList | The in vitro induction of corneal epithelial cells (CECs) from human induced pluripotent stem cells (iPSCs) represents a new strategy for obtaining CE stem/progenitor cells for the surgical reconstruction of a diseased or injured ocular surface. The clinical promise of this strategy is considerable, but if the approaches' potential is to be realised, robust methods for the purification of iPSC-derived CE lineage cells need to be developed to avoid contamination with other cells that may carry the risk of unwanted side effects, such as tumorigenesis. Experiments conducted here revealed that during CEC isolation, CD200-negative selection using a cell sorter considerably reduced the contamination of the cell population with various non-CECs compared with what could be achieved using TRA-1-60, a conventional negative marker for CECs. Furthermore, CD200-negative sorting did not affect the yield of CECs nor that of their stem/progenitor cells. Single-cell gene expression analysis for CEC sheets obtained using CD200-negative sorting showed that all analysed cells were CE-lineage cells, expressing PAX6, delta-N p63, and E-cadherin. Non-CECs, on the other hand, expressed non-CEC genes such as FGFR1 and RPE65. CD200, thus, represents a robust negative marker for purification of induced CE lineage cells, which is expressed by undifferentiated iPSCs and non-CECs, including iPSC-derived neural and retinal cells. Abstract The in vitro induction of corneal epithelial cells (CECs) from human induced pluripotent stem cells (iPSCs) represents a new strategy for obtaining CE stem/progenitor cells for the surgical reconstruction of a diseased or injured ocular surface. The clinical promise of this strategy is considerable, but if the approaches’ potential is to be realised, robust methods for the purification of iPSC-derived CE lineage cells need to be developed to avoid contamination with other cells that may carry the risk of unwanted side effects, such as tumorigenesis. Experiments conducted here revealed that during CEC isolation, CD200-negative selection using a cell sorter considerably reduced the contamination of the cell population with various non-CECs compared with what could be achieved using TRA-1-60, a conventional negative marker for CECs. Furthermore, CD200-negative sorting did not affect the yield of CECs nor that of their stem/progenitor cells. Single-cell gene expression analysis for CEC sheets obtained using CD200-negative sorting showed that all analysed cells were CE-lineage cells, expressing PAX6 , delta-N p63 , and E-cadherin . Non-CECs, on the other hand, expressed non-CEC genes such as FGFR1 and RPE65 . CD200, thus, represents a robust negative marker for purification of induced CE lineage cells, which is expressed by undifferentiated iPSCs and non-CECs, including iPSC-derived neural and retinal cells. The in vitro induction of corneal epithelial cells (CECs) from human induced pluripotent stem cells (iPSCs) represents a new strategy for obtaining CE stem/progenitor cells for the surgical reconstruction of a diseased or injured ocular surface. The clinical promise of this strategy is considerable, but if the approaches’ potential is to be realised, robust methods for the purification of iPSC-derived CE lineage cells need to be developed to avoid contamination with other cells that may carry the risk of unwanted side effects, such as tumorigenesis. Experiments conducted here revealed that during CEC isolation, CD200-negative selection using a cell sorter considerably reduced the contamination of the cell population with various non-CECs compared with what could be achieved using TRA-1-60, a conventional negative marker for CECs. Furthermore, CD200-negative sorting did not affect the yield of CECs nor that of their stem/progenitor cells. Single-cell gene expression analysis for CEC sheets obtained using CD200-negative sorting showed that all analysed cells were CE-lineage cells, expressing PAX6 , delta-N p63 , and E-cadherin . Non-CECs, on the other hand, expressed non-CEC genes such as FGFR1 and RPE65 . CD200, thus, represents a robust negative marker for purification of induced CE lineage cells, which is expressed by undifferentiated iPSCs and non-CECs, including iPSC-derived neural and retinal cells. |
ArticleNumber | 16550 |
Author | Katayama, Tomohiko Ishikawa, Yuki Hayashi, Ryuhei Nishida, Kohji Quantock, Andrew J |
Author_xml | – sequence: 1 givenname: Ryuhei surname: Hayashi fullname: Hayashi, Ryuhei email: ryuhei.hayashi@ophthal.med.osaka-u.ac.jp, ryuhei.hayashi@ophthal.med.osaka-u.ac.jp organization: Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka, 565-0871, Japan. ryuhei.hayashi@ophthal.med.osaka-u.ac.jp – sequence: 2 givenname: Yuki surname: Ishikawa fullname: Ishikawa, Yuki organization: Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka, 565-0871, Japan – sequence: 3 givenname: Tomohiko surname: Katayama fullname: Katayama, Tomohiko organization: Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka, 565-0871, Japan – sequence: 4 givenname: Andrew J surname: Quantock fullname: Quantock, Andrew J organization: Structural Biophysics Group, School of Optometry and Vision Sciences, College of Biomedical and Life Sciences, Cardiff University, Cardiff, CF24 4HQ, Wales, UK – sequence: 5 givenname: Kohji surname: Nishida fullname: Nishida, Kohji email: knishida@ophthal.med.osaka-u.ac.jp organization: Department of Ophthalmology, Osaka University Graduate School of Medicine, Suita, Osaka, 565-0871, Japan. knishida@ophthal.med.osaka-u.ac.jp |
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References | J Zhang (34845_CR6) 2009; 104 H Kamao (34845_CR3) 2014; 2 CP Hamel (34845_CR18) 1993; 268 C Büttner (34845_CR20) 2004; 198 M Mallo (34845_CR19) 2013; 140 U Pirvola (34845_CR15) 2002; 35 Y-T Chen (34845_CR23) 1996; 93 Y Sasamoto (34845_CR32) 2016; 6 S Tohyama (34845_CR8) 2013; 12 M Hellstrom (34845_CR22) 1999; 126 R Hayashi (34845_CR7) 2012; 7 R Hayashi (34845_CR14) 2017; 12 S Tohyama (34845_CR9) 2016; 23 C Tang (34845_CR12) 2011; 29 R Hayashi (34845_CR13) 2016; 531 J Walshe (34845_CR16) 2000; 90 C Chaloin-Dufau (34845_CR31) 1990; 32 J Yu (34845_CR1) 2007; 318 H Uosaki (34845_CR11) 2011; 6 JM Sperger (34845_CR17) 2003; 100 M Ohyama (34845_CR29) 2006; 116 M Rodrigues (34845_CR33) 1987; 34 34845_CR34 JE Kloepper (34845_CR30) 2008; 17 S-Y Chen (34845_CR28) 2011; 17 SJ Kattman (34845_CR5) 2011; 8 T Miyazaki (34845_CR35) 2012; 3 BH Chon (34845_CR27) 2013; 4 BT Kawasaki (34845_CR25) 2007; 364 K Takahashi (34845_CR2) 2007; 131 D Doi (34845_CR10) 2014; 2 R Hayashi (34845_CR24) 2007; 25 H Kataoka (34845_CR21) 1997; 39 H Kawasaki (34845_CR4) 2002; 99 BT Kawasaki (34845_CR26) 2008; 29 |
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Snippet | The in vitro induction of corneal epithelial cells (CECs) from human induced pluripotent stem cells (iPSCs) represents a new strategy for obtaining CE... Abstract The in vitro induction of corneal epithelial cells (CECs) from human induced pluripotent stem cells (iPSCs) represents a new strategy for obtaining CE... The in vitro induction of corneal epithelial cells (CECs) from human induced pluripotent stem cells (iPSCs) represents a new strategy for obtaining CE... |
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StartPage | 16550 |
SubjectTerms | CD200 antigen Contamination Cornea E-cadherin Epithelial cells Fibroblast growth factor receptor 1 Gene expression Negative selection Neural stem cells Pax6 protein Pluripotency Progenitor cells Purification Reconstructive surgery Retina Stem cells Tumorigenesis |
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Title | CD200 facilitates the isolation of corneal epithelial cells derived from human pluripotent stem cells |
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