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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Frontiers Media S.A.
2020-10-01
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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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