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Mesenchymal stem cells express epidermal markers in an in vitro reconstructed human skin model

Introduction: In skin traumas, such as burns, epidermal homeostasis is affected, often requiring clinical approaches. Different therapeutic strategies can be used including transplantation, besides the use of synthetic or natural materials with allogeneic cells. In this context, tissue engineering i...

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Autores principales: Dos Santos, Jeniffer Farias, Freitas-Marchi, Bruna Letícia, Reigado, Gustavo Roncoli, de Assis, Silvia Romano, Maria Engler, Silvya Stuchi, Chambergo Alcalde, Felipe Santiago, Nunes, Viviane Abreu
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2023
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Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9878690/
https://www.ncbi.nlm.nih.gov/pubmed/36712971
http://dx.doi.org/10.3389/fcell.2022.1012637
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author Dos Santos, Jeniffer Farias
Freitas-Marchi, Bruna Letícia
Reigado, Gustavo Roncoli
de Assis, Silvia Romano
Maria Engler, Silvya Stuchi
Chambergo Alcalde, Felipe Santiago
Nunes, Viviane Abreu
author_facet Dos Santos, Jeniffer Farias
Freitas-Marchi, Bruna Letícia
Reigado, Gustavo Roncoli
de Assis, Silvia Romano
Maria Engler, Silvya Stuchi
Chambergo Alcalde, Felipe Santiago
Nunes, Viviane Abreu
author_sort Dos Santos, Jeniffer Farias
collection PubMed
description Introduction: In skin traumas, such as burns, epidermal homeostasis is affected, often requiring clinical approaches. Different therapeutic strategies can be used including transplantation, besides the use of synthetic or natural materials with allogeneic cells. In this context, tissue engineering is an essential tool for skin regeneration, and using mesenchymal stem cells (MSC) from the umbilical cord appears to be a promising strategy in regenerative medicine due to its renewal and differentiation potential and hypo immunogenicity. We evaluated the transdifferentiation of MSC from umbilical cord into keratinocytes in three-dimensional (3D) in vitro skin models, using dermal equivalents composed by type I collagen with dermal fibroblasts and a commercial porcine skin decellularized matrix, both cultured at air-liquid interface (ALI). Methods: The expression of epidermal proteins cytokeratins (CK) 5, 14 and 10, involucrin and filaggrin was investigated by real-time PCR and immunofluorescence, in addition to the activity of epidermal kallikreins (KLK) on the hydrolysis of fluorogenic substrates. Results and discussion: The cultivation of MSCs with differentiation medium on these dermal supports resulted in organotypic cultures characterized by the expression of the epidermal markers CK5, CK14, CK10 and involucrin, mainly on the 7(th) day of culture, and filaggrin at 10(th) day in ALI. Also, there was a 3-fold increase in the KLK activity in the epidermal equivalents composed by MSC induced to differentiate into keratinocytes compared to the control (MSC cultivated in the proliferation medium). Specifically, the use of collagen and fibroblasts resulted in a more organized MSC-based organotypic culture in comparison to the decellularized matrix. Despite the non-typical epithelium structure formed by MSC onto dermal equivalents, the expression of important epidermal markers in addition to the paracrine effects of these cells in skin may indicate its potential use to produce skin-based substitutes.
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spelling pubmed-98786902023-01-27 Mesenchymal stem cells express epidermal markers in an in vitro reconstructed human skin model Dos Santos, Jeniffer Farias Freitas-Marchi, Bruna Letícia Reigado, Gustavo Roncoli de Assis, Silvia Romano Maria Engler, Silvya Stuchi Chambergo Alcalde, Felipe Santiago Nunes, Viviane Abreu Front Cell Dev Biol Cell and Developmental Biology Introduction: In skin traumas, such as burns, epidermal homeostasis is affected, often requiring clinical approaches. Different therapeutic strategies can be used including transplantation, besides the use of synthetic or natural materials with allogeneic cells. In this context, tissue engineering is an essential tool for skin regeneration, and using mesenchymal stem cells (MSC) from the umbilical cord appears to be a promising strategy in regenerative medicine due to its renewal and differentiation potential and hypo immunogenicity. We evaluated the transdifferentiation of MSC from umbilical cord into keratinocytes in three-dimensional (3D) in vitro skin models, using dermal equivalents composed by type I collagen with dermal fibroblasts and a commercial porcine skin decellularized matrix, both cultured at air-liquid interface (ALI). Methods: The expression of epidermal proteins cytokeratins (CK) 5, 14 and 10, involucrin and filaggrin was investigated by real-time PCR and immunofluorescence, in addition to the activity of epidermal kallikreins (KLK) on the hydrolysis of fluorogenic substrates. Results and discussion: The cultivation of MSCs with differentiation medium on these dermal supports resulted in organotypic cultures characterized by the expression of the epidermal markers CK5, CK14, CK10 and involucrin, mainly on the 7(th) day of culture, and filaggrin at 10(th) day in ALI. Also, there was a 3-fold increase in the KLK activity in the epidermal equivalents composed by MSC induced to differentiate into keratinocytes compared to the control (MSC cultivated in the proliferation medium). Specifically, the use of collagen and fibroblasts resulted in a more organized MSC-based organotypic culture in comparison to the decellularized matrix. Despite the non-typical epithelium structure formed by MSC onto dermal equivalents, the expression of important epidermal markers in addition to the paracrine effects of these cells in skin may indicate its potential use to produce skin-based substitutes. Frontiers Media S.A. 2023-01-12 /pmc/articles/PMC9878690/ /pubmed/36712971 http://dx.doi.org/10.3389/fcell.2022.1012637 Text en Copyright © 2023 Dos Santos, Freitas-Marchi, Reigado, de Assis, Maria Engler, Chambergo Alcalde and Nunes. https://creativecommons.org/licenses/by/4.0/This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.
spellingShingle Cell and Developmental Biology
Dos Santos, Jeniffer Farias
Freitas-Marchi, Bruna Letícia
Reigado, Gustavo Roncoli
de Assis, Silvia Romano
Maria Engler, Silvya Stuchi
Chambergo Alcalde, Felipe Santiago
Nunes, Viviane Abreu
Mesenchymal stem cells express epidermal markers in an in vitro reconstructed human skin model
title Mesenchymal stem cells express epidermal markers in an in vitro reconstructed human skin model
title_full Mesenchymal stem cells express epidermal markers in an in vitro reconstructed human skin model
title_fullStr Mesenchymal stem cells express epidermal markers in an in vitro reconstructed human skin model
title_full_unstemmed Mesenchymal stem cells express epidermal markers in an in vitro reconstructed human skin model
title_short Mesenchymal stem cells express epidermal markers in an in vitro reconstructed human skin model
title_sort mesenchymal stem cells express epidermal markers in an in vitro reconstructed human skin model
topic Cell and Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9878690/
https://www.ncbi.nlm.nih.gov/pubmed/36712971
http://dx.doi.org/10.3389/fcell.2022.1012637
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