Effects of fibroblasts and microenvironment on epidermal regeneration and tissue function in long-term skin equivalents.

Effects of fibroblasts and microenvironment on epidermal regeneration and tissue function in long-term skin equivalents. Research Abstract Details 

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Effects of fibroblasts and microenvironment on epidermal regeneration and tissue function in long-term skin equivalents. Abstract Text:

Karsten Boehnke,Nicolae Mirancea,Alessandra Pavesio,Norbert E Fusenig,Petra Boukamp, Stark,Karsten Boehnke,Nicolae Mirancea,Alessandra Pavesio,Norbert E Fusenig,Petra Boukamp, Stark,Karsten Boehnke,Nicolae Mirancea,Alessandra Pavesio,Norbert E Fusenig,Petra Boukamp, Stark,

In vitro generated skin models find growing interest as promising tools in basic research and clinical application in regenerative medicine. Here, we present further details of an improved long-term skin equivalent (SE) enabling mechanistic studies on skin reconstruction and epidermal function. Growth conditions of fibroblasts in a 3D scaffold were analysed to optimise the dermal microenvironment by providing an authentic dermal matrix for regular tissue reconstruction and function of cocultured keratinocytes. These SEs demonstrate sustained epidermal viability - over 12 weeks - with regular differentiation as substantiated by in vivo-like patterns of all differentiation products, exemplified here by the cornified envelope components loricrin and repetin. The continuous expression of all major tight junction components in the granular layer, shown here for ZO-1 in coherence with the presence of epidermal barrier lipids, and ultrastructural accumulation of lamellar bodies, collectively indicate proper epidermal barrier structures. Remarkably, cocultured keratinocytes exerted an ongoing proliferation-stimulating effect on fibroblasts colonising the scaffold comparable to a cocktail of fibroblast growth factors. Consequently, precultivation of dermal equivalents (DEs) in basal or growth factor-enriched media had only minor effects on the quality of epidermal regeneration in cocultures. As to the role of fibroblast numbers, complete absence of dermal cells resulted in atrophic epithelia but the effect of cell numbers as low as 5x10(4)cells/cm(2) on epidermal tissue quality equalled that of the standard density (2x10(5)cells/cm(2)). Surprisingly, precultivation of fibroblasts in the DEs for 7 days (standard) showed no better effect on epidermal tissue reformation as compared to 2 days whereas a precultivation period of 14 days resulted in atrophic epidermal and dermal tissue development. These data demonstrate, (i) the strict dependence of epidermal tissue regeneration on the presence of fibroblasts, (ii) the mutual keratinocyte-fibroblast interactions for cell proliferation and organogenesis, and (iii) the importance of the proper microenvironment for epidermal tissue function and supposedly for establishment of a stem cell niche in vitro.

Effects of fibroblasts and microenvironment on epidermal regeneration and tissue function in long-term skin equivalents. Publishing Authors By Initials

K Boehnke,N Mirancea,A Pavesio,NE Fusenig,P Boukamp,HJ Stark,K Boehnke,N Mirancea,A Pavesio,NE Fusenig,P Boukamp,HJ Stark,K Boehnke,N Mirancea,A Pavesio,NE Fusenig,P Boukamp,HJ Stark,

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Effects of fibroblasts and microenvironment on epidermal regeneration and tissue function in long-term skin equivalents. Journal Published:

PUBLICATION TYPE: Journal Article

Journal: European journal of cell biology

VOLUME: 86

Page Numbers: 731-46

Journal Abbreviation: Eur. J. Cell Biol.

ISSN: 0171-9335

DAY: 9

MONTH: 02

YEAR: 2007

Effects of fibroblasts and microenvironment on epidermal regeneration and tissue function in long-term skin equivalents. Information

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LANGUAGE: eng

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AFFILIATION: Division of Genetics of Skin Carcinogenesis, German Cancer Research Center (DKFZ), Heidelberg, Germany.

Country: Germany

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MEDLINETA: Eur J Cell Biol

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