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Identification of label‐retaining endothelial cells in adult human corneas: a new clue for the existence of endothelial stem cells
Author(s) -
HA THI BM,
HE Z,
CAMPOLMI N,
PISELLI S,
GAIN P,
PEOC'H M,
DUMOLLARD JM,
ACQUART S,
GARRAUD O,
THURET G
Publication year - 2012
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2012.2236.x
Subject(s) - stem cell , progenitor cell , microbiology and biotechnology , endothelial stem cell , corneal endothelium , endothelium , biology , in vivo , in vitro , immunology , chemistry , biochemistry , endocrinology , genetics
Purpose The lack of self‐renewal capacity of human corneal endothelial cells (EC) in vivo was explained by cell cycle arrest in the G1‐phase due to cell contact inhibition, TGF‐beta signaling, and stress induced premature senescence. Nevertheless, their residual ability to divide in primary culture suggests the existence of progenitor cells, probably located at the endothelial periphery (Whikehart.MolVis2005; He.StemCells2012). Stem cells are slow‐cycling cells characterized by their quiescent state in niches and their ability to retain for a long time markers of S‐Phase like BrDU or EdU. Aim: to search for the presence of label‐retaining EC in human corneas Methods Label retaining EC were searched by 5‐Ethynyl‐2’‐Deoxyuridine (Click‐it EdU) incorporation during long‐term culture: 30 days for organ cultured human corneas (n=10) or 15 days for in vitro primary cultured EC (n=5), both followed by a 30‐day culture without EdU. Flat‐mouted corneas and EC cultures were observed with an inverted fluorescent microscope Results Label‐retaining EC were observed in the peripheral area of all OC corneas, varying from 1 to 50. Numerous label‐retaining EC were also present in all primary cultures, always attached to Descemet membrane fragments Conclusion The presence of label‐retaining EC constitutes a new clue for the existence of corneal endothelial stem cells in human. Their apparent scarcity is consistent with the inability of the human corneal endothelium to repair in vivo, but isolation and expansion of these endothelial stem cells or progenitors could allow development of bioengineered endothelium