Comparative Effect of MSC Secretome to MSC Co-culture on Cardiomyocyte Gene Expression Under Hypoxic Conditions in vitro

IntroductionDespite major leaps in regenerative medicine, the regeneration of cardiomyocytes after ischemic conditions remains to elucidate. It is crucial to understand hypoxia induced cellular mechanisms to provide advanced treatment options, including the use of stem cell paracrine factors for myo...

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Main Authors: Nina Kastner, Julia Mester-Tonczar, Johannes Winkler, Denise Traxler, Andreas Spannbauer, Beate M. Rüger, Georg Goliasch, Noemi Pavo, Mariann Gyöngyösi, Katrin Zlabinger
Format: Article
Language:English
Published: Frontiers Media S.A. 2020-10-01
Series:Frontiers in Bioengineering and Biotechnology
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Online Access:https://www.frontiersin.org/article/10.3389/fbioe.2020.502213/full
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author Nina Kastner
Julia Mester-Tonczar
Johannes Winkler
Denise Traxler
Andreas Spannbauer
Beate M. Rüger
Georg Goliasch
Noemi Pavo
Mariann Gyöngyösi
Katrin Zlabinger
author_facet Nina Kastner
Julia Mester-Tonczar
Johannes Winkler
Denise Traxler
Andreas Spannbauer
Beate M. Rüger
Georg Goliasch
Noemi Pavo
Mariann Gyöngyösi
Katrin Zlabinger
author_sort Nina Kastner
collection DOAJ
description IntroductionDespite major leaps in regenerative medicine, the regeneration of cardiomyocytes after ischemic conditions remains to elucidate. It is crucial to understand hypoxia induced cellular mechanisms to provide advanced treatment options, including the use of stem cell paracrine factors for myocardial regeneration.Materials and MethodsIn this study, the regenerative potential of hypoxic human cardiomyocytes (group Hyp-CMC) in vitro was evaluated when co-cultured with human bone-marrow derived MSC (group Hyp-CMC-MSC) or stimulated with the secretome of MSC (group Hyp-CMC-SMSC). The secretome of normoxic MSC and CMC, and the hypoxic CMC was analyzed with a cytokine panel. Gene expression changes of HIF-1α, proliferation marker Ki-67 and cytokinesis marker RhoA over different reoxygenation time periods of 4, 8, 24, 48, and 72 h were analyzed in comparison to normoxic CMC and MSC. Further, the proinflammatory cytokine IL-18 protein expression change, metabolic activity and proliferation was assessed in all experimental setups.Results and ConclusionHIF-1α was persistently overexpressed in Hyp-CMC-SMSC as compared to Hyp-CMC (except at 72 h). Hyp-CMC-MSC showed a weaker HIF-1α expression than Hyp-CMC-SMSC in most tested time points, except after 8 h. The Ki-67 expression showed the strongest upregulation in Hyp-CMC after 24 and 48 h incubation, then returned to baseline level, while a temporary increase in Ki-67 expression in Hyp-CMC-MSC at 4 and 8 h and at 48 h in Hyp-CMC-SMSC could be observed. RhoA was increased in normoxic MSCs and in Hyp-CMC-SMSC over time, but not in Hyp-CMC-MSC. A temporary increase in IL-18 protein expression was detected in Hyp-CMC-SMSC and Hyp-CMC. Our study demonstrates timely dynamic changes in expression of different ischemia and regeneration-related genes of CMCs, depending from the culture condition, with stronger expression of HIF-1α, RhoA and IL-18 if the hypoxic CMC were subjected to the secretome of MSCs.
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spelling doaj.art-79685fd8ed864589baea4244251209aa2022-12-22T01:05:10ZengFrontiers Media S.A.Frontiers in Bioengineering and Biotechnology2296-41852020-10-01810.3389/fbioe.2020.502213502213Comparative Effect of MSC Secretome to MSC Co-culture on Cardiomyocyte Gene Expression Under Hypoxic Conditions in vitroNina Kastner0Julia Mester-Tonczar1Johannes Winkler2Denise Traxler3Andreas Spannbauer4Beate M. Rüger5Georg Goliasch6Noemi Pavo7Mariann Gyöngyösi8Katrin Zlabinger9Department of Cardiology, Medical University of Vienna, Vienna, AustriaDepartment of Cardiology, Medical University of Vienna, Vienna, AustriaDepartment of Cardiology, Medical University of Vienna, Vienna, AustriaDepartment of Cardiology, Medical University of Vienna, Vienna, AustriaDepartment of Cardiology, Medical University of Vienna, Vienna, AustriaDepartment of Blood Group Serology and Transfusion Medicine, Medical University of Vienna, Vienna, AustriaDepartment of Cardiology, Medical University of Vienna, Vienna, AustriaDepartment of Cardiology, Medical University of Vienna, Vienna, AustriaDepartment of Cardiology, Medical University of Vienna, Vienna, AustriaDepartment of Cardiology, Medical University of Vienna, Vienna, AustriaIntroductionDespite major leaps in regenerative medicine, the regeneration of cardiomyocytes after ischemic conditions remains to elucidate. It is crucial to understand hypoxia induced cellular mechanisms to provide advanced treatment options, including the use of stem cell paracrine factors for myocardial regeneration.Materials and MethodsIn this study, the regenerative potential of hypoxic human cardiomyocytes (group Hyp-CMC) in vitro was evaluated when co-cultured with human bone-marrow derived MSC (group Hyp-CMC-MSC) or stimulated with the secretome of MSC (group Hyp-CMC-SMSC). The secretome of normoxic MSC and CMC, and the hypoxic CMC was analyzed with a cytokine panel. Gene expression changes of HIF-1α, proliferation marker Ki-67 and cytokinesis marker RhoA over different reoxygenation time periods of 4, 8, 24, 48, and 72 h were analyzed in comparison to normoxic CMC and MSC. Further, the proinflammatory cytokine IL-18 protein expression change, metabolic activity and proliferation was assessed in all experimental setups.Results and ConclusionHIF-1α was persistently overexpressed in Hyp-CMC-SMSC as compared to Hyp-CMC (except at 72 h). Hyp-CMC-MSC showed a weaker HIF-1α expression than Hyp-CMC-SMSC in most tested time points, except after 8 h. The Ki-67 expression showed the strongest upregulation in Hyp-CMC after 24 and 48 h incubation, then returned to baseline level, while a temporary increase in Ki-67 expression in Hyp-CMC-MSC at 4 and 8 h and at 48 h in Hyp-CMC-SMSC could be observed. RhoA was increased in normoxic MSCs and in Hyp-CMC-SMSC over time, but not in Hyp-CMC-MSC. A temporary increase in IL-18 protein expression was detected in Hyp-CMC-SMSC and Hyp-CMC. Our study demonstrates timely dynamic changes in expression of different ischemia and regeneration-related genes of CMCs, depending from the culture condition, with stronger expression of HIF-1α, RhoA and IL-18 if the hypoxic CMC were subjected to the secretome of MSCs.https://www.frontiersin.org/article/10.3389/fbioe.2020.502213/fullhuman cardiomyocyteshypoxiaregenerationcell therapyMSC secretome
spellingShingle Nina Kastner
Julia Mester-Tonczar
Johannes Winkler
Denise Traxler
Andreas Spannbauer
Beate M. Rüger
Georg Goliasch
Noemi Pavo
Mariann Gyöngyösi
Katrin Zlabinger
Comparative Effect of MSC Secretome to MSC Co-culture on Cardiomyocyte Gene Expression Under Hypoxic Conditions in vitro
Frontiers in Bioengineering and Biotechnology
human cardiomyocytes
hypoxia
regeneration
cell therapy
MSC secretome
title Comparative Effect of MSC Secretome to MSC Co-culture on Cardiomyocyte Gene Expression Under Hypoxic Conditions in vitro
title_full Comparative Effect of MSC Secretome to MSC Co-culture on Cardiomyocyte Gene Expression Under Hypoxic Conditions in vitro
title_fullStr Comparative Effect of MSC Secretome to MSC Co-culture on Cardiomyocyte Gene Expression Under Hypoxic Conditions in vitro
title_full_unstemmed Comparative Effect of MSC Secretome to MSC Co-culture on Cardiomyocyte Gene Expression Under Hypoxic Conditions in vitro
title_short Comparative Effect of MSC Secretome to MSC Co-culture on Cardiomyocyte Gene Expression Under Hypoxic Conditions in vitro
title_sort comparative effect of msc secretome to msc co culture on cardiomyocyte gene expression under hypoxic conditions in vitro
topic human cardiomyocytes
hypoxia
regeneration
cell therapy
MSC secretome
url https://www.frontiersin.org/article/10.3389/fbioe.2020.502213/full
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