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|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|
LeGrand, Catherine F.
Kinnear, Beverley F.
Sobota, Radoslaw M.
Dye, Danielle E.
Lane, E. Birgitte
Coombe, Deirdre R.
|Published in:||Scientific Reports|
|Publisher / Ed. Institution:||Nature Publishing Group|
|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.|
|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, 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.
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