ORIGINAL RESEARCH article

Front. Cell Dev. Biol.

Sec. Stem Cell Research

Volume 13 - 2025 | doi: 10.3389/fcell.2025.1604605

This article is part of the Research TopicMetabolic Regulation of Stem Cell FateView all 3 articles

Comparative Cardiomyocyte Differentiation Potential of Rat Adipose-Derived Mesenchymal Stem Cells from Two Anatomical Sites: Metabolomic Profiling and Pathway Analysis

Provisionally accepted
  • 1Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
  • 2Department of Theriogenology, Faculty of Veterinary Medicine, Cairo University, Giza, Beni Suef, Egypt
  • 3Department of neurophysiology, National Center of Neurology and Psychiatry, Tokyo 187-8551, Japan., Tokyo, Japan
  • 4Faculty of Veterinary Medicine, Suez Canal University, Ismailia, Ismailia, Egypt
  • 5Juntendo University Urayasu Hospital, Urayasu, Chiba, Japan

The final, formatted version of the article will be published soon.

AbstractBackground: Adipose-derived mesenchymal stem cells (AD-MSCs) have emerged as a promising source for cardiac regenerative therapy due to their multipotency and ease of isolation. However, the impact of anatomical origin on their cardiomyocyte differentiation potential remains unclear. Metabolic analysis provides valuable real-time insights into the cellular metabolic state, capturing dynamic changes in metabolite concentrations that reflect both internal cellular mechanisms and external stimuli. This approach allows us to identify specific metabolic pathways activated during cardiomyocyte differentiation, offering a deeper understanding of how the anatomical origin of stem cells influences their differentiation potential and metabolic flexibility. Such insights are critical for optimizing stem cell-based therapies for cardiac regeneration.This study aimed to compare the differentiation capacity of AD-MSCs derived from peri-ovarian and peri-renal adipose tissue, with a focus on metabolic adaptations during cardiomyocyte differentiation.Methods: AD-MSCs were isolated from peri-ovarian and peri-renal fat of Sprague-Dawley rats and characterized by morphology, immunophenotyping, and multilineage differentiation potential. Cardiomyocyte differentiation was induced using 5-azacytidine, and morphological changes were assessed via phase-contrast microscopy and immunofluorescence staining for cardiac troponin T (cTnT). Untargeted metabolomic profiling was performed using gas chromatography-mass spectrometry (GC-MS), followed by principal component analysis (PCA), partial least squares discriminant analysis (PLS-DA), and pathway enrichment analysis.Results: Both peri-ovarian and peri-renal AD-MSCs exhibited similar fibroblast-like morphology, MSC-specific marker expression (CD44, CD90, CD29), and multilineage differentiation potential. Following cardiomyocyte induction, both groups displayed morphological changes indicative of differentiation and strong cTnT expression. Metabolomic analysis of the cardiogenic differentiation samples identified distinct metabolic adaptations between the two AD-MSC sources. Peri-ovarian AD-MSCs exhibited a broader metabolic reprogramming, with increased engagement of glycolysis, fructose metabolism, glycerolipid metabolism, and the TCA cycle, suggesting enhanced metabolic flexibility and energy efficiency. In contrast, peri-renal AD-MSCs relied more on galactose metabolism, indicating an alternative energy strategy during differentiation.Conclusion: The anatomical origin of AD-MSCs influences their metabolic landscape during cardiomyocyte differentiation. Peri-ovarian AD-MSCs demonstrated greater metabolic adaptability, potentially favoring their differentiation capacity, making them a promising candidate for cardiac regenerative applications.

Keywords: Adipose-derived mesenchymal stem cells, cardiomyocyte differentiation, 5azacytidine, Metabolomic profiling, Regenerative Medicine

Received: 02 Apr 2025; Accepted: 04 Jun 2025.

Copyright: © 2025 Farag, Samir, Ngeun, Kaneda, Hendawy, Takahashi and Tanaka. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) or licensor are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

* Correspondence:
Ahmed Farag, Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan
Ryou Tanaka, Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan

Disclaimer: All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.