Making more matrix : enhancing the deposition of dermal-epidermal junction components in vitro and accelerating organotypic skin culture development, using macromolecular crowding

Benny, Paula; Badowski, Cedric; Lane, E. Birgitte; Raghunath, Michael

doi:10.1089/ten.TEA.2013.0784

Publication type:	Article in scientific journal
Type of review:	Peer review (publication)
Title:	Making more matrix : enhancing the deposition of dermal-epidermal junction components in vitro and accelerating organotypic skin culture development, using macromolecular crowding
Authors:	Benny, Paula Badowski, Cedric Lane, E. Birgitte Raghunath, Michael
DOI:	10.1089/ten.TEA.2013.0784
Published in:	Tissue Engineering - Part A
Volume(Issue):	21
Issue:	1-2
Issue Date:	2015
Publisher / Ed. Institution:	Mary Ann Liebert
ISSN:	1937-335X 1937-3341
Language:	English
Subject (DDC):	571: Physiology and related subjects 616: Internal medicine and diseases
Abstract:	Skin is one of the most accessible tissues for experimental biomedical sciences, and cultured skin cells represent one of the longest-running clinical applications of stem cell therapy. However, culture-generated skin mimetic multicellular structures are still limited in their application by the time taken to develop these constructs in vitro and by their incomplete differentiation. The development of a functional dermal–epidermal junction (DEJ) is one of the most sought after aspects of cultured skin, and one of the hardest to recreate in vitro. At the DEJ, dermal fibroblasts and epidermal keratinocytes interact to form an interlinked basement membrane of extracellular matrix (ECM), which forms as a concerted action of both keratinocytes and fibroblasts. Successful formation of this basement membrane is essential for take and stability of cultured skin autografts. We studied interactive matrix production by monocultures and cocultures of primary human keratinocytes and fibroblasts in an attempt to improve the efficiency of basement membrane production in culture using mixed macromolecular crowding (mMMC); resulting ECM were enriched with the deposition of collagens I, IV, fibronectin, and laminin 332 (laminin 5) and also in collagen VII, the anchoring fibril component. Our in vitro data point to fibroblasts, rather than keratinocytes, as the major cellular contributors of the DEJ. Not only did we find more collagen VII production and deposition by fibroblasts in comparison to keratinocytes, but also observed that decellularized fibroblast ECM stimulated the production and deposition of collagen VII by keratinocytes, over and above that of keratinocyte monocultures. In confrontation cultures, keratinocytes and fibroblasts showed spontaneous segregation and demarcation of cell boundaries by DEJ protein deposition. Finally, mMMC was used in a classical organotypic coculture protocol with keratinocytes seeded over fibroblast-containing collagen gels. Applied during the submerged phase, mMMC was sufficient to accelerate the emergence of collagen VII along the de novo DEJ, together with stronger transglutaminase activity in the neoepidermis. Our findings corroborate the role of fibroblasts as important players in producing collagen VII and inducing collagen VII deposition in the DEJ, and that macromolecular crowding leads to organotypic epidermal differentiation in tissue culture in a significantly condensed time frame.
URI:	https://digitalcollection.zhaw.ch/handle/11475/12192
Fulltext version:	Published version
License (according to publishing contract):	Licence according to publishing contract
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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Benny, P., Badowski, C., Lane, E. B., & Raghunath, M. (2015). Making more matrix : enhancing the deposition of dermal-epidermal junction components in vitro and accelerating organotypic skin culture development, using macromolecular crowding. Tissue Engineering - Part A, 21(1-2). https://doi.org/10.1089/ten.TEA.2013.0784

Benny, P. et al. (2015) ‘Making more matrix : enhancing the deposition of dermal-epidermal junction components in vitro and accelerating organotypic skin culture development, using macromolecular crowding’, Tissue Engineering - Part A, 21(1-2). Available at: https://doi.org/10.1089/ten.TEA.2013.0784.

P. Benny, C. Badowski, E. B. Lane, and M. Raghunath, “Making more matrix : enhancing the deposition of dermal-epidermal junction components in vitro and accelerating organotypic skin culture development, using macromolecular crowding,” Tissue Engineering - Part A, vol. 21, no. 1-2, 2015, doi: 10.1089/ten.TEA.2013.0784.

BENNY, Paula, Cedric BADOWSKI, E. Birgitte LANE und Michael RAGHUNATH, 2015. Making more matrix : enhancing the deposition of dermal-epidermal junction components in vitro and accelerating organotypic skin culture development, using macromolecular crowding. Tissue Engineering - Part A. 2015. Bd. 21, Nr. 1-2. DOI 10.1089/ten.TEA.2013.0784

Benny, Paula, Cedric Badowski, E. Birgitte Lane, and Michael Raghunath. 2015. “Making More Matrix : Enhancing the Deposition of Dermal-Epidermal Junction Components in Vitro and Accelerating Organotypic Skin Culture Development, Using Macromolecular Crowding.” Tissue Engineering - Part A 21 (1-2). https://doi.org/10.1089/ten.TEA.2013.0784.

Benny, Paula, et al. “Making More Matrix : Enhancing the Deposition of Dermal-Epidermal Junction Components in Vitro and Accelerating Organotypic Skin Culture Development, Using Macromolecular Crowding.” Tissue Engineering - Part A, vol. 21, no. 1-2, 2015, https://doi.org/10.1089/ten.TEA.2013.0784.