The Different Binding Properties of Cultured Human Corneal Endothelial Cell Subpopulations to Descemet's Membrane Components.
To clarify the adherent properties of cultured human corneal endothelial cell (cHCEC) subpopulations (SPs).



Each SP was prepared by controlling the culture conditions or by using magnetic cell separation, and then confirmed by staining with several cell-surface markers. Binding abilities of HCEC SPs were examined by adding the cells to culture plates immobilized with collagens, laminins, or proteoglycans, and then centrifuging the plates. Adhered cells were then evaluated by phase-contrast microscopy.



The cHCECs were bound to laminin-511, laminin-411, and Type-IV collagen in a concentration-dependent manner, yet weakly bound to Perlecan, Agrin, and TSP-1. Comparison of the influence of cell-suspension vehicles on cHCEC attachment showed that cells suspended in Opti-MEM-I or Opeguard-MA were bound to laminin, yet no binding was observed in cells suspended in BSS-Plus. Next, we compared the adherent properties of HCEC SPs. Both the fully differentiated, mature cHCEC SP and the epithelial-to-mesenchymal-transitioned (EMT)-phenotype SP were found to attach to laminin- or collagen-coated plates. Interestingly, the binding properties to laminins differed among those SPs. Although the level of cells adhered to the laminin-411-coated plate was the same among the cHCEC SPs, the fully differentiated, mature cHCEC SP was significantly more tightly bound to laminin-511 than was the EMT-phenotype SP.



The findings of this study suggest that the binding ability of cHCECs to major Descemet's membrane components is distinct among cHCEC SPs, and that Opti-MEM-I and Opeguard-MA are useful cell-suspension vehicles for cell-injection therapy.
Department of Frontier Medical Science and Technology for Ophthalmology Kyoto Prefectural University of Medicine, Kyoto, Japan.
Munetoyo
M
Toda
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Morio
M
Ueno
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan 3Department of Ophthalmology, Meiji University of Integrative Medicine, Kyoto, Japan.
Jun
J
Yamada
Department of Frontier Medical Science and Technology for Ophthalmology Kyoto Prefectural University of Medicine, Kyoto, Japan.
Asako
A
Hiraga
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Hiroshi
H
Tanaka
Department of Ophthalmology, University Hospital Erlangen, Erlangen, Germany.
Ursula
U
Schlötzer-Schrehardt
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Chie
C
Sotozono
Department of Frontier Medical Science and Technology for Ophthalmology Kyoto Prefectural University of Medicine, Kyoto, Japan.
Shigeru
S
Kinoshita
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
Junji
J
Hamuro
D002448:Cell Adhesion; D002478:Cells, Cultured; D024141:Collagen Type IV / Q000378:metabolism*; D003886:Descemet Membrane / Q000166:cytology* / Q000378:metabolism; D004728:Endothelium, Corneal / Q000166:cytology* / Q000378:metabolism; D005434:Flow Cytometry; D006801:Humans; D007150:Immunohistochemistry; D039421:Integrin alpha2 / Q000096:biosynthesis; D007797:Laminin / Q000378:metabolism*; D008858:Microscopy, Phase-Contrast
D024141:Collagen Type IV; D039421:Integrin alpha2; D007797:Laminin