Gene Expression in Amnion-Derived Cells Cultured on Recombinant Laminin 332—A Preliminary Study

Human amniotic cells (hAC) exhibit characteristics of undifferentiated cells and immunomodulatory properties. Recognition of the relationship between amniotic cells and components of the extracellular matrix is an important condition for their ex vivo preparation and further successful clinical appl...

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Autores principales:	Katarzyna Skowron-Kandzia, Marcin Tomsia, Halina Koryciak-Komarska, Danuta Plewka, Patrycja Wieczorek, Piotr Czekaj
Formato:	article
Lenguaje:	EN
Publicado:	Frontiers Media S.A. 2021
Materias:	human amniotic cells extracellular matrix basement membrane laminin 332 integrins gene expression Medicine (General) R5-920
Acceso en línea:	https://doaj.org/article/fc0f79f70bd349cca1f5b1ef745ad24f
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spelling	oai:doaj.org-article:fc0f79f70bd349cca1f5b1ef745ad24f2021-11-10T05:55:01ZGene Expression in Amnion-Derived Cells Cultured on Recombinant Laminin 332—A Preliminary Study2296-858X10.3389/fmed.2021.719899https://doaj.org/article/fc0f79f70bd349cca1f5b1ef745ad24f2021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fmed.2021.719899/fullhttps://doaj.org/toc/2296-858XHuman amniotic cells (hAC) exhibit characteristics of undifferentiated cells and immunomodulatory properties. Recognition of the relationship between amniotic cells and components of the extracellular matrix is an important condition for their ex vivo preparation and further successful clinical application in regenerative medicine and transplantology. Laminin 332 (LN-332), as a natural component of the basement membrane of amniotic epithelial cells and a ligand for integrin receptors, may strongly influence the phenotype and fate of amniotic cells. We investigated the impact of recombinant LN-332 on hAC viability and expression of markers for pluripotency, early differentiation, adhesion, and immunomodulatory properties. During 14 days of culture, hAC were quantified and qualified by light microscopy, immunohistochemistry, immunocytochemistry, and flow cytometry. Gene expression was assessed with real-time polymerase chain reaction (RT-PCR) arrays and compared with differentiated cells originated from the three germ layers. LN-332 caused an over 2-fold increase in the total number of hAC, accompanied by a 75% reduction of SSEA-4-positive cells and an increase in HLA-ABC-positive cells. In particular, we observed that the presence of laminin 332 in the medium of a short-time culture modifies the effect of culture duration on hAC, enhancing time-dependent inhibition of expression of certain genes, including pluripotency and differentiation markers, laminin 332 subunits (which may be part of self-regulation of LN-332 synthesis by amniotic cells), and integrins. The changes observed in hAC were more distinct with respect to differentiated mesenchymal cells, resulting in more comparable phenotypes than those represented by differentiated endo- and ectodermal cells. We concluded that laminin 332 present in the culture medium influences to a certain extent proliferation, adhesion, and differentiation of amniotic cells in culture.Katarzyna Skowron-KandziaMarcin TomsiaHalina Koryciak-KomarskaDanuta PlewkaPatrycja WieczorekPiotr CzekajFrontiers Media S.A.articlehuman amniotic cellsextracellular matrixbasement membranelaminin 332integrinsgene expressionMedicine (General)R5-920ENFrontiers in Medicine, Vol 8 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	human amniotic cells extracellular matrix basement membrane laminin 332 integrins gene expression Medicine (General) R5-920
spellingShingle	human amniotic cells extracellular matrix basement membrane laminin 332 integrins gene expression Medicine (General) R5-920 Katarzyna Skowron-Kandzia Marcin Tomsia Halina Koryciak-Komarska Danuta Plewka Patrycja Wieczorek Piotr Czekaj Gene Expression in Amnion-Derived Cells Cultured on Recombinant Laminin 332—A Preliminary Study
description	Human amniotic cells (hAC) exhibit characteristics of undifferentiated cells and immunomodulatory properties. Recognition of the relationship between amniotic cells and components of the extracellular matrix is an important condition for their ex vivo preparation and further successful clinical application in regenerative medicine and transplantology. Laminin 332 (LN-332), as a natural component of the basement membrane of amniotic epithelial cells and a ligand for integrin receptors, may strongly influence the phenotype and fate of amniotic cells. We investigated the impact of recombinant LN-332 on hAC viability and expression of markers for pluripotency, early differentiation, adhesion, and immunomodulatory properties. During 14 days of culture, hAC were quantified and qualified by light microscopy, immunohistochemistry, immunocytochemistry, and flow cytometry. Gene expression was assessed with real-time polymerase chain reaction (RT-PCR) arrays and compared with differentiated cells originated from the three germ layers. LN-332 caused an over 2-fold increase in the total number of hAC, accompanied by a 75% reduction of SSEA-4-positive cells and an increase in HLA-ABC-positive cells. In particular, we observed that the presence of laminin 332 in the medium of a short-time culture modifies the effect of culture duration on hAC, enhancing time-dependent inhibition of expression of certain genes, including pluripotency and differentiation markers, laminin 332 subunits (which may be part of self-regulation of LN-332 synthesis by amniotic cells), and integrins. The changes observed in hAC were more distinct with respect to differentiated mesenchymal cells, resulting in more comparable phenotypes than those represented by differentiated endo- and ectodermal cells. We concluded that laminin 332 present in the culture medium influences to a certain extent proliferation, adhesion, and differentiation of amniotic cells in culture.
format	article
author	Katarzyna Skowron-Kandzia Marcin Tomsia Halina Koryciak-Komarska Danuta Plewka Patrycja Wieczorek Piotr Czekaj
author_facet	Katarzyna Skowron-Kandzia Marcin Tomsia Halina Koryciak-Komarska Danuta Plewka Patrycja Wieczorek Piotr Czekaj
author_sort	Katarzyna Skowron-Kandzia
title	Gene Expression in Amnion-Derived Cells Cultured on Recombinant Laminin 332—A Preliminary Study
title_short	Gene Expression in Amnion-Derived Cells Cultured on Recombinant Laminin 332—A Preliminary Study
title_full	Gene Expression in Amnion-Derived Cells Cultured on Recombinant Laminin 332—A Preliminary Study
title_fullStr	Gene Expression in Amnion-Derived Cells Cultured on Recombinant Laminin 332—A Preliminary Study
title_full_unstemmed	Gene Expression in Amnion-Derived Cells Cultured on Recombinant Laminin 332—A Preliminary Study
title_sort	gene expression in amnion-derived cells cultured on recombinant laminin 332—a preliminary study
publisher	Frontiers Media S.A.
publishDate	2021
url	https://doaj.org/article/fc0f79f70bd349cca1f5b1ef745ad24f
work_keys_str_mv	AT katarzynaskowronkandzia geneexpressioninamnionderivedcellsculturedonrecombinantlaminin332apreliminarystudy AT marcintomsia geneexpressioninamnionderivedcellsculturedonrecombinantlaminin332apreliminarystudy AT halinakoryciakkomarska geneexpressioninamnionderivedcellsculturedonrecombinantlaminin332apreliminarystudy AT danutaplewka geneexpressioninamnionderivedcellsculturedonrecombinantlaminin332apreliminarystudy AT patrycjawieczorek geneexpressioninamnionderivedcellsculturedonrecombinantlaminin332apreliminarystudy AT piotrczekaj geneexpressioninamnionderivedcellsculturedonrecombinantlaminin332apreliminarystudy
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Gene Expression in Amnion-Derived Cells Cultured on Recombinant Laminin 332—A Preliminary Study

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