RRC ID 54723
Author Hayashi R, Ishikawa Y, Katayama T, Quantock AJ, Nishida K.
Title CD200 facilitates the isolation of corneal epithelial cells derived from human pluripotent stem cells.
Journal Sci Rep
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.
Volume 8(1)
Pages 16550
Published 2018-11-8
DOI 10.1038/s41598-018-34845-2
PII 10.1038/s41598-018-34845-2
PMID 30410112
PMC PMC6224558
MeSH Antigens, CD / genetics Antigens, CD / metabolism* Antigens, Surface / genetics Cadherins / genetics Cell Differentiation Cell Lineage Cells, Cultured Epithelium, Corneal / cytology* Epithelium, Corneal / immunology Humans Induced Pluripotent Stem Cells / cytology* Induced Pluripotent Stem Cells / immunology PAX6 Transcription Factor / genetics Pluripotent Stem Cells / cytology Pluripotent Stem Cells / immunology Proteoglycans / genetics Single-Cell Analysis
IF 4.011
Times Cited 3
Human and Animal Cells 201B7(HPS0063)