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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.
Afiliación
  • Sasamoto Y; Division of Genetics, Brigham and Women's Hospital, Boston, MA, USA; Transplant Research Program, Boston Children's Hospital, Boston, MA, USA.
  • Yeung PC; Division of Medical Sciences, Harvard Medical School, Boston, MA, USA; Department of Surgery, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, Hong Kong SAR, China.
  • Tran J; Transplant Research Program, Boston Children's Hospital, Boston, MA, USA.
  • Frank MH; Transplant Research Program, Boston Children's Hospital, Boston, MA, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA; Harvard Skin Disease Research Center, Department of Dermatology, Brigham and Women's Hospital, Boston, MA, USA; School of Medical and Health Sciences, Edith
  • Frank NY; Division of Genetics, Brigham and Women's Hospital, Boston, MA, USA; Harvard Stem Cell Institute, Harvard University, Cambridge, MA, USA; Department of Medicine, VA Boston Healthcare System, Boston, MA, USA. Electronic address: nyfrank@bwh.harvard.edu.
STAR Protoc ; 4(3): 102503, 2023 Sep 15.
Article en En | MEDLINE | ID: mdl-37669162
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.
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Texto completo: 1 Base de datos: MEDLINE Asunto principal: Limbo de la Córnea / Epitelio Corneal Límite: Humans Idioma: En Revista: STAR Protoc Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos
Texto completo
Imprimir
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Texto completo: 1 Base de datos: MEDLINE Asunto principal: Limbo de la Córnea / Epitelio Corneal Límite: Humans Idioma: En Revista: STAR Protoc Año: 2023 Tipo del documento: Article País de afiliación: Estados Unidos