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Isolation of human photoreceptor precursors via a cell surface marker panel from stem cell-derived retinal organoids and fetal retinae

Isolation of human photoreceptor precursors via a cell surface marker panel from stem cell-derived retinal organoids and fetal retinae
Isolation of human photoreceptor precursors via a cell surface marker panel from stem cell-derived retinal organoids and fetal retinae
Loss of photoreceptor cells due to retinal degeneration is one of the main causes of blindness in the developed world. Although there is currently no effective treatment, cell replacement therapy using stem-cell-derived photoreceptor cells may be a feasible future treatment option. In order to ensure safety and efficacy of this approach, robust cell isolation and purification protocols must be developed. To this end, we previously developed a biomarker panel for the isolation of mouse photoreceptor precursors from the developing mouse retina and mouse embryonic stem cell cultures. In the current study we applied this approach to the human pluripotent stem cell (hPSC) system, and identified novel biomarker combinations that can be leveraged for the isolation of human photoreceptors. Human retinal samples and hPSC-derived retinal organoid cultures were screened against 242 human monoclonal antibodies using a high through-put flow cytometry approach. We identified 46 biomarkers with significant expression levels in the human retina and hPSC differentiation cultures. Human retinal cell samples, either from fetal tissue or derived from embryonic and induced pluripotent stem cell cultures, were fluorescence-activated cell sorted (FACS) using selected candidate biomarkers that showed expression in discrete cell populations. Enrichment for photoreceptors and exclusion of mitotically active cells was demonstrated by immunocytochemical analysis with photoreceptor-specific antibodies and Ki-67. We established a biomarker combination, which enables the robust purification of viable human photoreceptors from both human retinae and hPSC-derived organoid cultures.
1066-5099
709-722
Lakowski, Jorn
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Welby, Emily
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Budinger, Dimitri
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Di Marco, Fabiana
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Di Foggia, Valentina
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Bainbridge, James W.B.
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Wallace, Kyle
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Gamm, David M.
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Ali, Robin R.
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Sowden, Jane C.
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Lakowski, Jorn
1856e739-982a-412a-87c7-abf1610f5384
Welby, Emily
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Budinger, Dimitri
3c9b43aa-f25c-4639-ad0b-9ba524b5b01c
Di Marco, Fabiana
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Di Foggia, Valentina
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Bainbridge, James W.B.
be3077b2-2b9c-41de-9f1c-ecb6dda9ad82
Wallace, Kyle
ffbf7b7a-2a07-41fd-b82a-f04999f38bdb
Gamm, David M.
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Ali, Robin R.
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Sowden, Jane C.
e042184a-dbe3-4a58-9f91-c31a5463b893

Lakowski, Jorn, Welby, Emily, Budinger, Dimitri, Di Marco, Fabiana, Di Foggia, Valentina, Bainbridge, James W.B., Wallace, Kyle, Gamm, David M., Ali, Robin R. and Sowden, Jane C. (2018) Isolation of human photoreceptor precursors via a cell surface marker panel from stem cell-derived retinal organoids and fetal retinae. Stem Cells, 36 (5), 709-722. (doi:10.1002/stem.2775).

Record type: Article

Abstract

Loss of photoreceptor cells due to retinal degeneration is one of the main causes of blindness in the developed world. Although there is currently no effective treatment, cell replacement therapy using stem-cell-derived photoreceptor cells may be a feasible future treatment option. In order to ensure safety and efficacy of this approach, robust cell isolation and purification protocols must be developed. To this end, we previously developed a biomarker panel for the isolation of mouse photoreceptor precursors from the developing mouse retina and mouse embryonic stem cell cultures. In the current study we applied this approach to the human pluripotent stem cell (hPSC) system, and identified novel biomarker combinations that can be leveraged for the isolation of human photoreceptors. Human retinal samples and hPSC-derived retinal organoid cultures were screened against 242 human monoclonal antibodies using a high through-put flow cytometry approach. We identified 46 biomarkers with significant expression levels in the human retina and hPSC differentiation cultures. Human retinal cell samples, either from fetal tissue or derived from embryonic and induced pluripotent stem cell cultures, were fluorescence-activated cell sorted (FACS) using selected candidate biomarkers that showed expression in discrete cell populations. Enrichment for photoreceptors and exclusion of mitotically active cells was demonstrated by immunocytochemical analysis with photoreceptor-specific antibodies and Ki-67. We established a biomarker combination, which enables the robust purification of viable human photoreceptors from both human retinae and hPSC-derived organoid cultures.

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More information

Accepted/In Press date: 14 December 2017
e-pub ahead of print date: 12 January 2018
Published date: May 2018

Identifiers

Local EPrints ID: 425643
URI: http://eprints.soton.ac.uk/id/eprint/425643
ISSN: 1066-5099
PURE UUID: 3db5811f-a4f1-4b47-8713-4c50b2dfabd3
ORCID for Jorn Lakowski: ORCID iD orcid.org/0000-0003-4214-7580

Catalogue record

Date deposited: 30 Oct 2018 17:30
Last modified: 13 Nov 2021 03:01

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Contributors

Author: Jorn Lakowski ORCID iD
Author: Emily Welby
Author: Dimitri Budinger
Author: Fabiana Di Marco
Author: Valentina Di Foggia
Author: James W.B. Bainbridge
Author: Kyle Wallace
Author: David M. Gamm
Author: Robin R. Ali
Author: Jane C. Sowden

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