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Title
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Fish scale-derived scaffolds for culturing human corneal endothelial cells
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Author
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Abstract
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| Purpose. To investigate the biocompatibility of fish scale-derived scaffolds (FSS) with primary human corneal endothelial cells (HCEnCs). Methods. HCEnCs were isolated from 30 donor corneas in a donor-matched study and plated in precoated Lab-Tek slides (n = 15) and FSS (n = 15). Cell morphology, proliferation/migration, and glucose uptake were studied (n = 30). Hoechst, ethidium homodimer, and calcein AM (NEC) staining was performed to determine viability and toxicity (n = 6). The cell surface area was calculated based on calcein AM staining. HCEnCs were stained for ZO-1 (n = 6) to detect tight junctions and to measure cell morphology; Ki-67 (n = 6) to measure proliferating cells; and vinculin to quantify focal adhesions (n = 6). The formation of de novo extracellular matrix was analyzed using histology (n = 6). Results. HCEnCs attach and grow faster on Lab-Tek slides compared to the undulating topography of the FSS. At day 11, HCEnCs on Lab-Tek slide grew 100% confluent, while FSS was only 65% confluent (p = 0.0883), with no significant difference in glucose uptake between the two (p = 0.5181) (2.2 mu g/mL in Lab-Tek versus 2.05 mu g/mL in FSS). HEC staining showed no toxicity. The surface area of the cells in Lab-Tek was 409.1 mu m(2) compared to 452.2 mu m(2) on FSS, which was not significant (p = 0.5325). ZO-1 showed the presence of tight junctions in both conditions; however, hexagonality was higher (74% in Lab-Tek versus 45% in FSS; p = 0.0006) with significantly less polymorphic cells on Lab-Tek slides (8% in Lab-Tek versus 16% in FSS; p = 0.0041). Proliferative cells were detected in both conditions (4.6% in Lab-Tek versus 4.2% in FSS; p = 0.5922). Vinculin expression was marginally higher in HCEnCs cultured on Lab-Tek (234 versus 199 focal adhesions; p = 0.0507). Histological analysis did not show the formation of a basement membrane. Conclusions. HCEnCs cultured on precoated FSS form a monolayer, displaying correct morphology, cytocompatibility, and absence of toxicity. FSS needs further modification in terms of structure and surface chemistry before considering it as a potential carrier for cultured HCEnCs. |
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Language
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English
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Source (journal)
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Stem Cells International
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Publication
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2018
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ISSN
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1687-966X
1687-9678
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Volume/pages
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(2018)
, 11 p.
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Article Reference
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8146834
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ISI
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000432116100001
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Pubmed ID
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29853917
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Medium
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E-only publicatie
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Full text (Publisher's DOI)
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Full text (open access)
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