Please use this identifier to cite or link to this item: https://hdl.handle.net/10356/154013
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dc.contributor.authorSeidelmann, Ninaen_US
dc.contributor.authorDuarte Campos, Daniela F.en_US
dc.contributor.authorRohde, Malenaen_US
dc.contributor.authorJohnen, Sandraen_US
dc.contributor.authorSalla, Sabineen_US
dc.contributor.authorYam, Gary Hin-Faien_US
dc.contributor.authorMehta, Jodhbir Singhen_US
dc.contributor.authorWalter, Peteren_US
dc.contributor.authorFuest, Matthiasen_US
dc.date.accessioned2022-06-07T06:01:49Z-
dc.date.available2022-06-07T06:01:49Z-
dc.date.issued2021-
dc.identifier.citationSeidelmann, N., Duarte Campos, D. F., Rohde, M., Johnen, S., Salla, S., Yam, G. H., Mehta, J. S., Walter, P. & Fuest, M. (2021). Human platelet lysate as a replacement for fetal bovine serum in human corneal stromal keratocyte and fibroblast culture. Journal of Cellular and Molecular Medicine, 25(20), 9647-9659. https://dx.doi.org/10.1111/jcmm.16912en_US
dc.identifier.issn1582-1838en_US
dc.identifier.urihttps://hdl.handle.net/10356/154013-
dc.description.abstractThe isolation and propagation of primary human corneal stromal keratocytes (CSK) are crucial for cellular research and corneal tissue engineering. However, this delicate cell type easily transforms into stromal fibroblasts (SF) and scar inducing myofibroblasts (Myo-SF). Current protocols mainly rely on xenogeneic fetal bovine serum (FBS). Human platelet lysate (hPL) could be a viable, potentially autologous, alternative. We found high cell survival with both supplements in CSK and SF. Cell numbers and Ki67+ ratios increased with higher fractions of hPL and FBS in CSK and SF. We detected a loss in CSK marker expression (Col8A2, ALDH3A1 and LUM) with increasing fractions of FBS and hPL in CSK and SF. The expression of the Myo-SF marker SMA increased with higher amounts of FBS but decreased with incremental hPL substitution in both cell types, implying an antifibrotic effect of hPL. Immunohistochemistry confirmed the RT-PCR findings. bFGF and HGF were only found in hPL and could be responsible for suppressing the Myo-SF conversion. Considering all findings, we propose 0.5% hPL as a suitable substitution in CSK culture, as this xeno-free component efficiently preserved CSK characteristics, with non-inferiority in terms of cell viability, cell number and proliferation in comparison to the established 0.5% FBS protocol.en_US
dc.language.isoenen_US
dc.relation.ispartofJournal of Cellular and Molecular Medicineen_US
dc.rights© 2021 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.en_US
dc.subjectEngineering::Materialsen_US
dc.titleHuman platelet lysate as a replacement for fetal bovine serum in human corneal stromal keratocyte and fibroblast cultureen_US
dc.typeJournal Articleen
dc.contributor.schoolSchool of Materials Science and Engineeringen_US
dc.contributor.organizationSingapore Eye Research Instituteen_US
dc.contributor.organizationSingapore National Eye Centreen_US
dc.contributor.organizationDuke-National University of Singaporeen_US
dc.identifier.doi10.1111/jcmm.16912-
dc.description.versionPublished versionen_US
dc.identifier.pmid34486211-
dc.identifier.scopus2-s2.0-85114285559-
dc.identifier.issue20en_US
dc.identifier.volume25en_US
dc.identifier.spage9647en_US
dc.identifier.epage9659en_US
dc.subject.keywordsCell Cultureen_US
dc.subject.keywordsCorneaen_US
item.fulltextWith Fulltext-
item.grantfulltextopen-
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