In vitro expansion of keratinocytes on human dermal fibroblast-derived matrix retains their stem-like characteristics

Wong, Chee-Wai; LeGrand, Catherine F.; Kinnear, Beverley F.; Sobota, Radoslaw M.; Ramalingam, Rajkumar; Dye, Danielle E.; Raghunath, Michael; Lane, E. Birgitte; Coombe, Deirdre R.

doi:10.1038/s41598-019-54793-9

Please use this identifier to cite or link to this item: https://doi.org/10.21256/zhaw-22188

Publication type:	Article in scientific journal
Type of review:	Peer review (publication)
Title:	In vitro expansion of keratinocytes on human dermal fibroblast-derived matrix retains their stem-like characteristics
Authors:	Wong, Chee-Wai LeGrand, Catherine F. Kinnear, Beverley F. Sobota, Radoslaw M. Ramalingam, Rajkumar Dye, Danielle E. Raghunath, Michael Lane, E. Birgitte Coombe, Deirdre R.
et. al:	No
DOI:	10.1038/s41598-019-54793-9 10.21256/zhaw-22188
Published in:	Scientific Reports
Volume(Issue):	9
Issue:	1
Page(s):	18561
Issue Date:	6-Dec-2019
Publisher / Ed. Institution:	Nature Publishing Group
ISSN:	2045-2322
Language:	English
Subjects:	Cell culture technique; Cell differentiation; Cell proliferation; Cultured cells; Coculture technique; Dermis; Extracellular matrix; Feeder cells; Fibroblasts; Human; Keratinocytes; Skin transplantation
Subject (DDC):	610.28: Biomedicine, biomedical engineering
Abstract:	The long-term expansion of keratinocytes under conditions that avoid xenogeneic components (i.e. animal serum- and feeder cell-free) generally causes diminished proliferation and increased terminal differentiation. Here we present a culture system free of xenogeneic components that retains the self-renewal capacity of primary human keratinocytes. In vivo the extracellular matrix (ECM) of the tissue microenvironment has a major influence on a cell's fate. We used ECM from human dermal fibroblasts, cultured under macromolecular crowding conditions to facilitate matrix deposition and organisation, in a xenogeneic-free keratinocyte expansion protocol. Phospholipase A2 decellularisation produced ECM whose components resembled the core matrix composition of natural dermis by proteome analyses. Keratinocytes proliferated rapidly on these matrices, retained their small size, expressed p63, lacked keratin 10 and rarely expressed keratin 16. The colony forming efficiency of these keratinocytes was enhanced over that of keratinocytes grown on collagen I, indicating that dermal fibroblast-derived matrices maintain the in vitro expansion of keratinocytes in a stem-like state. Keratinocyte sheets formed on such matrices were multi-layered with superior strength and stability compared to the single-layered sheets formed on collagen I. Thus, keratinocytes expanded using our xenogeneic-free protocol retained a stem-like state, but when triggered by confluence and calcium concentration, they stratified to produce epidermal sheets with a potential clinical use.
URI:	https://digitalcollection.zhaw.ch/handle/11475/22188
Fulltext version:	Published version
License (according to publishing contract):	CC BY 4.0: Attribution 4.0 International
Departement:	Life Sciences and Facility Management
Organisational Unit:	Institute of Chemistry and Biotechnology (ICBT)
Appears in collections:	Publikationen Life Sciences und Facility Management

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Wong, C.-W., LeGrand, C. F., Kinnear, B. F., Sobota, R. M., Ramalingam, R., Dye, D. E., Raghunath, M., Lane, E. B., & Coombe, D. R. (2019). In vitro expansion of keratinocytes on human dermal fibroblast-derived matrix retains their stem-like characteristics. Scientific Reports, 9(1), 18561. https://doi.org/10.1038/s41598-019-54793-9

Wong, C.-W. et al. (2019) ‘In vitro expansion of keratinocytes on human dermal fibroblast-derived matrix retains their stem-like characteristics’, Scientific Reports, 9(1), p. 18561. Available at: https://doi.org/10.1038/s41598-019-54793-9.

C.-W. Wong et al., “In vitro expansion of keratinocytes on human dermal fibroblast-derived matrix retains their stem-like characteristics,” Scientific Reports, vol. 9, no. 1, p. 18561, Dec. 2019, doi: 10.1038/s41598-019-54793-9.

WONG, Chee-Wai, Catherine F. LEGRAND, Beverley F. KINNEAR, Radoslaw M. SOBOTA, Rajkumar RAMALINGAM, Danielle E. DYE, Michael RAGHUNATH, E. Birgitte LANE und Deirdre R. COOMBE, 2019. In vitro expansion of keratinocytes on human dermal fibroblast-derived matrix retains their stem-like characteristics. Scientific Reports. 6 Dezember 2019. Bd. 9, Nr. 1, S. 18561. DOI 10.1038/s41598-019-54793-9

Wong, Chee-Wai, Catherine F. LeGrand, Beverley F. Kinnear, Radoslaw M. Sobota, Rajkumar Ramalingam, Danielle E. Dye, Michael Raghunath, E. Birgitte Lane, and Deirdre R. Coombe. 2019. “In Vitro Expansion of Keratinocytes on Human Dermal Fibroblast-Derived Matrix Retains Their Stem-like Characteristics.” Scientific Reports 9 (1): 18561. https://doi.org/10.1038/s41598-019-54793-9.

Wong, Chee-Wai, et al. “In Vitro Expansion of Keratinocytes on Human Dermal Fibroblast-Derived Matrix Retains Their Stem-like Characteristics.” Scientific Reports, vol. 9, no. 1, Dec. 2019, p. 18561, https://doi.org/10.1038/s41598-019-54793-9.