In-Vitro Effects of Secreted Frizzled-Related Protein 1 (SFRP1) On Human Corneal Epithelial Cells

You, J; Munoz-Erazo, L; Wen, L; Hodge, C; Madigan, MC; Sutton, G

In-Vitro Effects of Secreted Frizzled-Related Protein 1 (SFRP1) On Human Corneal Epithelial Cells

You, J Munoz-Erazo, L Wen, L Hodge, C

Madigan, MC Sutton, G

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Publication Type:: Journal Article
Citation:: Current Eye Research, 2018, 43 (4), pp. 455 - 459
Issue Date:: 2018-04-03

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Field	Value	Language
dc.contributor.author	You, J	en_US
dc.contributor.author	Munoz-Erazo, L	en_US
dc.contributor.author	Wen, L	en_US
dc.contributor.author	Hodge, C https://orcid.org/0000-0002-0516-6486	en_US
dc.contributor.author	Madigan, MC	en_US
dc.contributor.author	Sutton, G	en_US
dc.date.issued	2018-04-03	en_US
dc.identifier.citation	Current Eye Research, 2018, 43 (4), pp. 455 - 459	en_US
dc.identifier.issn	0271-3683	en_US
dc.identifier.uri	http://hdl.handle.net/10453/133932
dc.description.abstract	© 2018 Taylor & Francis Group, LLC. Purpose: Limbal corneal epithelial cells (LCECs) are responsible for corneal epithelial cell regeneration. However, corneal central epithelial cells (CCECs) are also suggested to display potential for self-renewal. Additionally, a better understanding of molecules that regulate corneal epithelial cell regeneration is important for studying conditions affecting the cornea, for example, keratoconus. Given our previous findings of reduced levels of secreted frizzled-related protein 1 (SFRP1) in tears from keratoconus patients compared to controls, we investigated the effects of SFRP1 on the proliferation and survival of cultured central and limbal human corneal epithelial cells. Material and methods: Limbal and central corneal explants were established from postmortem human corneas, and cultured in CnT-PR, an epithelial-specific tissue culture media. Subcultured cells from explants were immunostained for the cytokeratins CK3, 12, 19, and the proliferative/oligopotent markers Ki67 and p63. BrdU flow cytometry, Alamar Blue and LDH assays were used to assess effects of SFRP1 treatment on central and LCECs. Results: Primary limbal and central corneal epithelial cells were successfully cultured in vitro to confluence (P6 and P4, respectively). They all expressed varying levels of cytokeratins CK3, CK12 and CK19, and Ki67 and p63. Additionally, they showed significantly increased metabolic activity after SFRP1 treatment (p < 0.05), with a maximum response at 1 μg/mL of SPRF1. No difference in proliferation was detected in SFRP1 treated LCECs; however, a reduction in cell death was noted (p < 0.05). Conclusion: Similar to the LCECs, primary human CCECs can be cultured in vitro, and expressed epithelial markers. SFRP1 demonstrated an improvement on the metabolic activity of both CCECs and LCECs, which in LCECs could be resulted from reduced cell death. This may have implications in degenerative corneal disorders, such as keratoconus.	en_US
dc.relation.ispartof	Current Eye Research	en_US
dc.relation.isbasedon	10.1080/02713683.2018.1431284	en_US
dc.subject.classification	Ophthalmology & Optometry	en_US
dc.subject.mesh	Epithelium, Corneal	en_US
dc.subject.mesh	Limbus Corneae	en_US
dc.subject.mesh	Cells, Cultured	en_US
dc.subject.mesh	Humans	en_US
dc.subject.mesh	Keratoconus	en_US
dc.subject.mesh	Intercellular Signaling Peptides and Proteins	en_US
dc.subject.mesh	Membrane Proteins	en_US
dc.subject.mesh	Immunohistochemistry	en_US
dc.subject.mesh	Cell Proliferation	en_US
dc.subject.mesh	Adult	en_US
dc.subject.mesh	Aged	en_US
dc.subject.mesh	Middle Aged	en_US
dc.subject.mesh	Female	en_US
dc.subject.mesh	Male	en_US
dc.title	In-Vitro Effects of Secreted Frizzled-Related Protein 1 (SFRP1) On Human Corneal Epithelial Cells	en_US
dc.type	Journal Article
utslib.citation.volume	4	en_US
utslib.citation.volume	43	en_US
utslib.for	1113 Opthalmology and Optometry	en_US
utslib.for	0601 Biochemistry and Cell Biology	en_US
pubs.embargo.period	Not known	en_US
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Graduate School of Health
utslib.copyright.status	closed_access
pubs.issue	4	en_US
pubs.publication-status	Published	en_US
pubs.volume	43	en_US

Abstract:

© 2018 Taylor & Francis Group, LLC. Purpose: Limbal corneal epithelial cells (LCECs) are responsible for corneal epithelial cell regeneration. However, corneal central epithelial cells (CCECs) are also suggested to display potential for self-renewal. Additionally, a better understanding of molecules that regulate corneal epithelial cell regeneration is important for studying conditions affecting the cornea, for example, keratoconus. Given our previous findings of reduced levels of secreted frizzled-related protein 1 (SFRP1) in tears from keratoconus patients compared to controls, we investigated the effects of SFRP1 on the proliferation and survival of cultured central and limbal human corneal epithelial cells. Material and methods: Limbal and central corneal explants were established from postmortem human corneas, and cultured in CnT-PR, an epithelial-specific tissue culture media. Subcultured cells from explants were immunostained for the cytokeratins CK3, 12, 19, and the proliferative/oligopotent markers Ki67 and p63. BrdU flow cytometry, Alamar Blue and LDH assays were used to assess effects of SFRP1 treatment on central and LCECs. Results: Primary limbal and central corneal epithelial cells were successfully cultured in vitro to confluence (P6 and P4, respectively). They all expressed varying levels of cytokeratins CK3, CK12 and CK19, and Ki67 and p63. Additionally, they showed significantly increased metabolic activity after SFRP1 treatment (p < 0.05), with a maximum response at 1 μg/mL of SPRF1. No difference in proliferation was detected in SFRP1 treated LCECs; however, a reduction in cell death was noted (p < 0.05). Conclusion: Similar to the LCECs, primary human CCECs can be cultured in vitro, and expressed epithelial markers. SFRP1 demonstrated an improvement on the metabolic activity of both CCECs and LCECs, which in LCECs could be resulted from reduced cell death. This may have implications in degenerative corneal disorders, such as keratoconus.

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http://hdl.handle.net/10453/133932