1. ASCs derived from burn patients are more prone to increased oxidative metabolism and reactive oxygen species upon passaging
- Author
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Tiffany C. Heard, Shanmugasundaram Natesan, Robert J. Christy, Belinda I. Gómez, Tony Chao, David M. Burmeister, Grace Chu Yuan Chu, and Linda E. Sousse
- Subjects
Mitochondrial ROS ,Medicine (General) ,Medicine (miscellaneous) ,Adipose tissue ,Burn ,QD415-436 ,Oxidative phosphorylation ,Mitochondrion ,Biochemistry ,Biochemistry, Genetics and Molecular Biology (miscellaneous) ,Andrology ,R5-920 ,Adipocytes ,Humans ,Medicine ,chemistry.chemical_classification ,Reactive oxygen species ,business.industry ,Stem Cells ,Research ,Cell Differentiation ,ROS ,Cell Biology ,Respirometry ,Adipose stem cells ,Mitochondria ,Oxidative Stress ,Adipose Tissue ,chemistry ,Hypermetabolism ,Molecular Medicine ,Stem cell ,Reactive Oxygen Species ,business ,Wound healing ,Glycolysis - Abstract
Background Patients with severe burn injury (over 20% of the total body surface area) experience profound hypermetabolism which significantly prolongs wound healing. Adipose-derived stem cells (ASCs) have been proposed as an attractive solution for treating burn wounds, including the potential for autologous ASC expansion. While subcutaneous adipocytes display an altered metabolic profile post-burn, it is not known if this is the case with the stem cells associated with the adipose tissue. Methods ASCs were isolated from discarded burn skin of severely injured human subjects (BH, n = 6) and unburned subcutaneous adipose tissue of patients undergoing elective abdominoplasty (UH, n = 6) and were analyzed at passages 2, 4, and 6. Flow cytometry was used to quantify ASC cell surface markers CD90, CD105, and CD73. Mitochondrial abundance and reactive oxygen species (ROS) production were determined with MitoTracker Green and MitoSOX Red, respectively, while JC-10 Mitochondrial Membrane Potential Assays were also performed. Mitochondrial respiration and glycolysis were analyzed with a high-resolution respirometer (Seahorse XFe24 Analyzer). Results There was no difference in age between BH and UH (34 ± 6 and 41 ± 4 years, respectively, P = 0.49). While passage 2 ASCs had lower ASC marker expression than subsequent passages, there were no significant differences in the expression between BH and UH ASCs. Similarly, no differences in mitochondrial abundance or membrane potential were found amongst passages or groups. Two-way ANOVA showed a significant effect (P P = 0.035) but not UH ASCs. On the contrary, basal glycolysis significantly decreased in BH ASCs (P = 0.011) with subsequent passaging, but not UH ASCs. Conclusions In conclusion, ASCs from burned individuals become increasingly oxidative and less glycolytic upon passaging when compared to ASCs from unburned patients. This increase in oxidative capacities was associated with ROS production in later passages. While the autologous expansion of ASCs holds great promise for treating burned patients with limited donor sites, the potential negative consequences of using them require further investigation.
- Published
- 2021