Metabolic maturation of differentiating cardiosphere-derived cells
Cardiosphere-derived cells (CDCs) can be expanded in vitro and induced to differentiate along the cardiac lineage. To recapitulate the phenotype of an adult cardiomyocyte, differentiating progenitors need to upregulate mitochondrial glucose and fatty acid oxidation. Here we cultured and differentiat...
| Main Authors: | , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Elsevier
2021-07-01
|
| Series: | Stem Cell Research |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S1873506121002683 |
| _version_ | 1819027494431358976 |
|---|---|
| author | Khadijeh Kathy Pakzad Jun Jie Tan Stephanie Anderson Mary Board Kieran Clarke Carolyn A. Carr |
| author_facet | Khadijeh Kathy Pakzad Jun Jie Tan Stephanie Anderson Mary Board Kieran Clarke Carolyn A. Carr |
| author_sort | Khadijeh Kathy Pakzad |
| collection | DOAJ |
| description | Cardiosphere-derived cells (CDCs) can be expanded in vitro and induced to differentiate along the cardiac lineage. To recapitulate the phenotype of an adult cardiomyocyte, differentiating progenitors need to upregulate mitochondrial glucose and fatty acid oxidation. Here we cultured and differentiated CDCs using protocols aimed to maintain stemness or to promote differentiation, including triggering fatty acid oxidation using an agonist of peroxisome proliferator-activated receptor alpha (PPARα). Metabolic changes were characterised in undifferentiated CDCs and during differentiation towards a cardiac phenotype. CDCs from rat atria were expanded on fibronectin or collagen IV via cardiosphere formation. Differentiation was assessed using flow cytometry and qPCR and substrate metabolism was quantified using radiolabelled substrates. Collagen IV promoted proliferation of CDCs whereas fibronectin primed cells for differentiation towards a cardiac phenotype. In both populations, treatment with 5-Azacytidine induced a switch towards oxidative metabolism, as shown by changes in gene expression, decreased glycolytic flux and increased oxidation of glucose and palmitate. Addition of a PPARα agonist during differentiation increased both glucose and fatty acid oxidation and expression of cardiac genes. We conclude that oxidative metabolism and cell differentiation act in partnership with increases in one driving an increase in the other. |
| first_indexed | 2024-12-21T05:43:22Z |
| format | Article |
| id | doaj.art-f0ec3124754045e0a60432db138d9e0e |
| institution | Directory Open Access Journal |
| issn | 1873-5061 |
| language | English |
| last_indexed | 2024-12-21T05:43:22Z |
| publishDate | 2021-07-01 |
| publisher | Elsevier |
| record_format | Article |
| series | Stem Cell Research |
| spelling | doaj.art-f0ec3124754045e0a60432db138d9e0e2022-12-21T19:14:12ZengElsevierStem Cell Research1873-50612021-07-0154102422Metabolic maturation of differentiating cardiosphere-derived cellsKhadijeh Kathy Pakzad0Jun Jie Tan1Stephanie Anderson2Mary Board3Kieran Clarke4Carolyn A. Carr5Department of Physiology, Anatomy & Genetics, University of Oxford, UKDepartment of Physiology, Anatomy & Genetics, University of Oxford, UK; Advanced Medical and Dental Institute, Universiti Sains Malaysia, Penang, MalaysiaDepartment of Physiology, Anatomy & Genetics, University of Oxford, UKDepartment of Physiology, Anatomy & Genetics, University of Oxford, UKDepartment of Physiology, Anatomy & Genetics, University of Oxford, UKDepartment of Physiology, Anatomy & Genetics, University of Oxford, UK; Corresponding author.Cardiosphere-derived cells (CDCs) can be expanded in vitro and induced to differentiate along the cardiac lineage. To recapitulate the phenotype of an adult cardiomyocyte, differentiating progenitors need to upregulate mitochondrial glucose and fatty acid oxidation. Here we cultured and differentiated CDCs using protocols aimed to maintain stemness or to promote differentiation, including triggering fatty acid oxidation using an agonist of peroxisome proliferator-activated receptor alpha (PPARα). Metabolic changes were characterised in undifferentiated CDCs and during differentiation towards a cardiac phenotype. CDCs from rat atria were expanded on fibronectin or collagen IV via cardiosphere formation. Differentiation was assessed using flow cytometry and qPCR and substrate metabolism was quantified using radiolabelled substrates. Collagen IV promoted proliferation of CDCs whereas fibronectin primed cells for differentiation towards a cardiac phenotype. In both populations, treatment with 5-Azacytidine induced a switch towards oxidative metabolism, as shown by changes in gene expression, decreased glycolytic flux and increased oxidation of glucose and palmitate. Addition of a PPARα agonist during differentiation increased both glucose and fatty acid oxidation and expression of cardiac genes. We conclude that oxidative metabolism and cell differentiation act in partnership with increases in one driving an increase in the other.http://www.sciencedirect.com/science/article/pii/S1873506121002683Extracellular matrixGlycolytic metabolismOxidative phosphorylationCardiac progenitor cellsCardiomyogenic differentiation |
| spellingShingle | Khadijeh Kathy Pakzad Jun Jie Tan Stephanie Anderson Mary Board Kieran Clarke Carolyn A. Carr Metabolic maturation of differentiating cardiosphere-derived cells Stem Cell Research Extracellular matrix Glycolytic metabolism Oxidative phosphorylation Cardiac progenitor cells Cardiomyogenic differentiation |
| title | Metabolic maturation of differentiating cardiosphere-derived cells |
| title_full | Metabolic maturation of differentiating cardiosphere-derived cells |
| title_fullStr | Metabolic maturation of differentiating cardiosphere-derived cells |
| title_full_unstemmed | Metabolic maturation of differentiating cardiosphere-derived cells |
| title_short | Metabolic maturation of differentiating cardiosphere-derived cells |
| title_sort | metabolic maturation of differentiating cardiosphere derived cells |
| topic | Extracellular matrix Glycolytic metabolism Oxidative phosphorylation Cardiac progenitor cells Cardiomyogenic differentiation |
| url | http://www.sciencedirect.com/science/article/pii/S1873506121002683 |
| work_keys_str_mv | AT khadijehkathypakzad metabolicmaturationofdifferentiatingcardiospherederivedcells AT junjietan metabolicmaturationofdifferentiatingcardiospherederivedcells AT stephanieanderson metabolicmaturationofdifferentiatingcardiospherederivedcells AT maryboard metabolicmaturationofdifferentiatingcardiospherederivedcells AT kieranclarke metabolicmaturationofdifferentiatingcardiospherederivedcells AT carolynacarr metabolicmaturationofdifferentiatingcardiospherederivedcells |