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Metabolic plasticity during transition to naïve-like pluripotency in canine embryo-derived stem cells
I C Tobias1, R R Isaac2, J G Dierolf1
1Department of Physiology and Pharmacology, Schulich School of Medicine & Dentistry, Western University, London, Ontario, Canada.
Stem Cell Research
|May 21, 2018
Summary
Canine pluripotent stem cells (PSCs) exhibit distinct metabolic states supporting their proliferation. Understanding these bioenergetic differences, particularly in naïve (2iL) versus primed (LIF-FGF2) states, can improve canine stem cell culture.
Area of Science:
- * Stem cell biology and regenerative medicine.
- * Cellular metabolism and bioenergetics.
- * Canine developmental biology.
Background:
- * Canine embryonic stem cells (cESCs) are valuable translational models but are understudied.
- * cESCs exist in primed and naïve pluripotent states, influenced by culture conditions (LIF-FGF2 vs. 2iL).
- * Proliferation rates of cESCs differ between states and are slower than in human or mouse PSCs.
Purpose of the Study:
- * To investigate the distinct bioenergetic mechanisms supporting proliferation in primed-like (LIF-FGF2) and naïve (2iL) cESCs.
- * To understand how metabolic pathways contribute to ATP generation and biomass accumulation in different cESC states.
- * To identify potential targets for improving canine PSC derivation and culture.
Main Methods:
- * Expansion of cESCs in LIF-FGF2 and 2iL conditions.
- * Measurement of respiratory capacity, mitochondrial membrane potential, and ATP levels.
- * Analysis of mitochondrial ultrastructure and cristae organization.
- * Metabolic stress assays involving inhibition of oxidative phosphorylation and glycolysis.
Main Results:
- * 2iL cESCs demonstrated higher respiratory capacity, altered mitochondrial stoichiometry, and increased mitochondrial polarization compared to LIF-FGF2 cESCs.
- * 2iL cESCs showed immature ultrastructural features, including novel cristae organization changes.
- * Enhanced ATP levels in 2iL cESCs correlated with altered retrograde signaling, while LIF-FGF2 cESCs displayed a lipogenic phenotype.
- * Oxidative phosphorylation inhibition impaired 2iL cESC proliferation and ATP production, whereas LIF-FGF2 cESCs remained sensitive to glycolysis inhibition.
Conclusions:
- * Distinct bioenergetic pathways fuel the proliferation of different canine pluripotent stem cell states.
- * Naïve (2iL) cESCs rely more on oxidative phosphorylation, while primed-like (LIF-FGF2) cESCs are more dependent on glycolysis.
- * These findings provide insights into optimizing canine PSC culture and regenerative medicine applications.
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