Protocol for isolating human BCAM-positive corneal progenitor cells by flow cytometry and cell sorting

BCAM-positive basal limbal epithelial cells are an early transit-amplifying cell population (TAC) capable of holoclone formation and corneal epithelial differentiation. Here, we present a protocol for isolating BCAM-positive cells from human donor corneas by flow cytometry and cell sorting. We descr...

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Detalles Bibliográficos
Autores principales: Sasamoto, Yuzuru, Yeung, Philip C., Tran, Johnathan, Frank, Markus H., Frank, Natasha Y.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Elsevier 2023
Materias:
Protocol
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10485628/
https://www.ncbi.nlm.nih.gov/pubmed/37669162
http://dx.doi.org/10.1016/j.xpro.2023.102503
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author Sasamoto, Yuzuru
Yeung, Philip C.
Tran, Johnathan
Frank, Markus H.
Frank, Natasha Y.
author_facet Sasamoto, Yuzuru
Yeung, Philip C.
Tran, Johnathan
Frank, Markus H.
Frank, Natasha Y.
author_sort Sasamoto, Yuzuru
collection PubMed
description BCAM-positive basal limbal epithelial cells are an early transit-amplifying cell population (TAC) capable of holoclone formation and corneal epithelial differentiation. Here, we present a protocol for isolating BCAM-positive cells from human donor corneas by flow cytometry and cell sorting. We describe steps for cell dissection and dissociation, antibody staining, and flow cytometry. We then detail procedures for culturing the purified BCAM-positive and BCAM-negative cells for holoclone and cell sheet formation assays to study the factors that regulate corneal regeneration. For complete details on the use and execution of this protocol, please refer to Sasamoto et al.(1)
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spelling pubmed-104856282023-09-09 Protocol for isolating human BCAM-positive corneal progenitor cells by flow cytometry and cell sorting Sasamoto, Yuzuru Yeung, Philip C. Tran, Johnathan Frank, Markus H. Frank, Natasha Y. STAR Protoc Protocol BCAM-positive basal limbal epithelial cells are an early transit-amplifying cell population (TAC) capable of holoclone formation and corneal epithelial differentiation. Here, we present a protocol for isolating BCAM-positive cells from human donor corneas by flow cytometry and cell sorting. We describe steps for cell dissection and dissociation, antibody staining, and flow cytometry. We then detail procedures for culturing the purified BCAM-positive and BCAM-negative cells for holoclone and cell sheet formation assays to study the factors that regulate corneal regeneration. For complete details on the use and execution of this protocol, please refer to Sasamoto et al.(1) Elsevier 2023-09-03 /pmc/articles/PMC10485628/ /pubmed/37669162 http://dx.doi.org/10.1016/j.xpro.2023.102503 Text en © 2023. https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Protocol
Sasamoto, Yuzuru
Yeung, Philip C.
Tran, Johnathan
Frank, Markus H.
Frank, Natasha Y.
Protocol for isolating human BCAM-positive corneal progenitor cells by flow cytometry and cell sorting
title Protocol for isolating human BCAM-positive corneal progenitor cells by flow cytometry and cell sorting
title_full Protocol for isolating human BCAM-positive corneal progenitor cells by flow cytometry and cell sorting
title_fullStr Protocol for isolating human BCAM-positive corneal progenitor cells by flow cytometry and cell sorting
title_full_unstemmed Protocol for isolating human BCAM-positive corneal progenitor cells by flow cytometry and cell sorting
title_short Protocol for isolating human BCAM-positive corneal progenitor cells by flow cytometry and cell sorting
title_sort protocol for isolating human bcam-positive corneal progenitor cells by flow cytometry and cell sorting
topic Protocol
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC10485628/
https://www.ncbi.nlm.nih.gov/pubmed/37669162
http://dx.doi.org/10.1016/j.xpro.2023.102503
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