Abstract
The relationship between mitochondrial metabolism and cell viability and differentiation in stem cells (SCs) remains poorly understood. In the present study, we compared mitochondrial physiology and metabolism between P19SCs before/after differentiation and present a unique fingerprint of the association between mitochondrial activity, cell differentiation and stemness. In comparison with their differentiated counterparts, pluripotency of P19SCs was correlated with a strong glycolytic profile and decreased mitochondrial biogenesis and complexity: round, low-polarized and inactive mitochondria with a closed permeability transition pore. This decreased mitochondrial capacity increased their resistance against dichloroacetate. Thus, stimulation of mitochondrial function by growing P19SCs in glutamine/pyruvate-containing medium reduced their glycolytic phenotype, induced loss of pluripotent potential, compromised differentiation and became P19SCs sensitive to dichloroacetate. Because of the central role of this type of SCs in teratocarcinoma development, our findings highlight the importance of mitochondrial metabolism in stemness, proliferation, differentiation and chemoresistance. In addition, the present work suggests the regulation of mitochondrial metabolism as a tool for inducing cell differentiation in stem line therapies.
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Abbreviations
- ANT:
-
adenine nucleotide translocase
- ADP:
-
adenosine diphosphate
- ATP5A:
-
ATP synthase subunit α
- COX IV:
-
cytochrome c oxidase subunit IV
- CSCs:
-
cancer stem cells
- CyP-D:
-
cyclophilin D
- ESCs:
-
embryonic stem cells
- ETC:
-
electron transport chain
- FCCP:
-
carbonyl cyanide 4-(trifluoromethoxy)phenylhydrazone
- HBSS:
-
Hank’s Balanced Salt Solution
- mPTP:
-
mitochondrial permeability transition pore
- MTCO1:
-
cytochrome c oxidase subunit I
- mTFA:
-
mitochondrial transcription factor A
- MTG:
-
MitoTracker Green
- MTR:
-
MitoTracker Red
- NANOG:
-
nanog homeobox protein
- NDUFB8:
-
NADH dehydrogenase (ubiquinone) 1 β subcomplex 8
- NMR:
-
nuclear magnetic resonance
- OCT4:
-
octamer-binding transcription factor
- OXPHOS:
-
oxidative phosphorylation
- P19dC:
-
differentiated P19 cell
- P19SC:
-
P19 stem cell
- PDH:
-
pyruvate dehydrogenase
- PDK:
-
pyruvate dehydrogenase kinase
- PGC-1:
-
peroxisomal proliferator-activated receptor coactivator 1
- PI:
-
propidium iodide
- RA:
-
retinoic acid
- ROS:
-
reactive oxygen species
- SCs:
-
stem cells
- SDHB:
-
succinate dehydrogenase (ubiquinone) iron-sulfur subunit
- SOX2:
-
sex determining region Y-box 2
- TMRM:
-
tetramethylrhodamine methyl ester
- TOM20:
-
translocase of outer mitochondrial membrane 20
- TROMA-1:
-
cytokeratin 8 Endo-A
- UQCRC2:
-
ubiquinol-cytochrome c reductase core protein II
- Δψm:
-
mitochondrial transmembrane electric potential
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Acknowledgements
We thank Isabel Nunes Correia and Luisa Cortes for technical assistance. This work was performed with funding from the Portuguese Foundation for Science and Technology (FCT) co-funded by COMPETE/FEDER/National Budget (PTDC/QUI-BIQ/101052/2008 and PEst-C/SAU/LA0001/2013-2014). Vega-Naredo I acknowledges a Marie Curie Intra-European Fellowship from the EU Seventh Framework Programme (PIEF-GA-2009-251850) and the FCT (SFRH/BPD/86534/2012). The FCT also supported Mesquita KA (SFRH/BD/66138/2009). The present work was also supported by QREN project # 4832, reference ‘CENTRO-07-ST24-FEDER-002008’.
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Vega-Naredo, I., Loureiro, R., Mesquita, K. et al. Mitochondrial metabolism directs stemness and differentiation in P19 embryonal carcinoma stem cells. Cell Death Differ 21, 1560–1574 (2014). https://doi.org/10.1038/cdd.2014.66
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DOI: https://doi.org/10.1038/cdd.2014.66
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