Exploring the Role of Anatomical Origin in the Cardiomyocyte Differentiation of Adipose-Derived Stem Cells: Metabolomic Profiling and Pathway Analysis

Ahmed Farag; Sai Ngeun; Masahiro Kaneda; Haney Samir; Ryou Tanaka

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Exploring the Role of Anatomical Origin in the Cardiomyocyte Differentiation of Adipose-Derived Stem Cells: Metabolomic Profiling and Pathway Analysis

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¹ Department of Veterinary Medicine, Faculty of Agriculture, Tokyo University of Agriculture and Technology
² Laboratory of Veterinary Diagnostic Imaging, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan
³ Laboratory of Veterinary Anatomy, Division of Animal Life Science, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan
⁴ Department of Theriogenology, Faculty of Veterinary Medicine, Cairo University, Giza, 12211, Egypt
⁵ Veterinary Teaching Hospital, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Tokyo 183-8509, Japan

Academic Editor: Serafino Fazio

Published: 09 May 2025 by MDPI in The 3rd International Electronic Conference on Biomedicines session Biomedicine in Cardiovascular Diseases

Abstract:

Abstract
Introduction: Adipose-derived mesenchymal stem cells (AD-MSCs) have emerged as a promising tool in regenerative medicine, particularly in cardiac repair. These cells are recognized for their capacity to differentiate into cardiomyocyte-like cells under specific conditions. However, the influence of their anatomical origin on their metabolic profiles and differentiation potential has not been fully elucidated. This study investigates AD-MSCs from two distinct anatomical locations—peri-ovarian and peri-renal adipose tissue—to determine how their origin impacts their ability to differentiate into cardiomyocyte-like cells.

Methods: AD-MSCs were isolated from peri-ovarian and peri-renal fat of female rats and characterized by their morphology, surface marker expression, and trilineage differentiation potential. Cardiomyocyte differentiation was induced using 5-azacytidine. Cellular responses were evaluated through morphological analysis, immunofluorescence staining for cardiac troponin T (cTnT), and untargeted metabolomic profiling using gas chromatography–mass spectrometry (GC-MS).

Results: Both peri-ovarian and peri-renal AD-MSCs displayed characteristic mesenchymal morphology, surface marker profiles, and multi-lineage differentiation potential. While both groups successfully differentiated into cardiomyocyte-like cells, metabolomic profiling revealed distinct metabolic adaptations. Peri-ovarian AD-MSCs demonstrated heightened activity in glycolysis, fructose metabolism, glycerolipid metabolism, and the TCA cycle, suggesting a more robust metabolic reprogramming suited for differentiation. In contrast, peri-renal AD-MSCs showed increased reliance on galactose metabolism. Immunofluorescence staining confirmed the expression of cTnT in differentiated cells from both groups.

Conclusions: These findings highlight the critical role of anatomical origin in shaping the metabolic and differentiation capabilities of AD-MSCs. Peri-ovarian AD-MSCs exhibited superior metabolic reprogramming, positioning them as a more favorable source for cardiac tissue engineering and regenerative applications.

Keywords: Adipose-derived mesenchymal stem cells (AD-MSCs), Cardiomyocyte differentiation, Metabolomics, Peri-ovarian, Peri-renal, 5-azacytidine.

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