Abstract

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

RAS ID

62016

Document Type

Journal Article

Date of Publication

9-15-2023

Volume

4

Issue

3

School

School of Medical and Health Sciences

Creative Commons License

Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License
This work is licensed under a Creative Commons Attribution-Noncommercial-No Derivative Works 4.0 License.

Publisher

Elsevier

Comments

Sasamoto, Y., Yeung, P. C., Tran, J., Frank, M. H., & Frank, N. Y. (2023). Protocol for isolating human BCAM-positive corneal progenitor cells by flow cytometry and cell sorting. STAR protocols, 4(3), 102503. https://doi.org/10.1016/j.xpro.2023.102503

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Link to publisher version (DOI)

10.1016/j.xpro.2023.102503