Human mesenchymal stem cells exhibit altered mitochondrial dynamics and poor survival in high glucose microenvironment

Ejlal Abu-El-Rub; Fatimah Almahasneh; Ramada R Khasawneh; Ayman Alzu'bi; Doaa Ghorab; Rawan Almazari; Huthaifa Magableh; Ahmad Sanajleh; Haitham Shlool; Mohammad Mazari; Noor S Bader; Joud Al-Momani

doi:10.4252/wjsc.v15.i12.1093

Human mesenchymal stem cells exhibit altered mitochondrial dynamics and poor survival in high glucose microenvironment

World J Stem Cells. 2023 Dec 26;15(12):1093-1103. doi: 10.4252/wjsc.v15.i12.1093.

Affiliations

¹ Department of Basic Medical Sciences, Faculty of Medicine, Yarmouk University, Irbid 21163, Jordan. ejlal.abuelrub@yu.edu.jo.
² Department of Basic Medical Sciences, Faculty of Medicine, Yarmouk University, Irbid 21163, Jordan.
³ Department of Pathology, Faculty of Medicine, Mansoura University, Mansoura 35516, Egypt.
⁴ Department of Basic Medical Sciences, Yarmouk University, Irbid 21163, Jordan.

Abstract

Background: Mesenchymal stem cells (MSCs) are a type of stem cells that possess relevant regenerative abilities and can be used to treat many chronic diseases. Diabetes mellitus (DM) is a frequently diagnosed chronic disease characterized by hyperglycemia which initiates many multisystem complications in the long-run. DM patients can benefit from MSCs transplantation to curb down the pathological consequences associated with hyperglycemia persistence and restore the function of damaged tissues. MSCs therapeutic outcomes are found to last for short period of time and ultimately these regenerative cells are eradicated and died in DM disease model.

Aim: To investigate the impact of high glucose or hyperglycemia on the cellular and molecular characteristics of MSCs.

Methods: Human adipose tissue-derived MSCs (hAD-MSCs) were seeded in low (5.6 mmol/L of glucose) and high glucose (25 mmol/L of glucose) for 7 d. Cytotoxicity, viability, mitochondrial dynamics, and apoptosis were deplored using specific kits. Western blotting was performed to measure the protein expression of phosphatidylinositol 3-kinase (PI3K), TSC1, and mammalian target of rapamycin (mTOR) in these cells.

Results: hAD-MSCs cultured in high glucose for 7 d demonstrated marked decrease in their viability, as shown by a significant increase in lactate dehydrogenase (P < 0.01) and a significant decrease in Trypan blue (P < 0.05) in these cells compared to low glucose control. Mitochondrial membrane potential, indicated by tetramethylrhodamine ethyl ester (TMRE) fluorescence intensity, and nicotinamide adenine dinucleotide (NAD+)/NADH ratio were significantly dropped (P < 0.05 for TMRE and P < 0.01 for NAD+/NADH) in high glucose exposed hAD-MSCs, indicating disturbed mitochondrial function. PI3K protein expression significantly decreased in high glucose culture MSCs (P < 0.05 compared to low glucose) and it was coupled with significant upregulation in TSC1 (P < 0.05) and downregulation in mTOR protein expression (P < 0.05). Mitochondrial complexes I, IV, and V were downregulated profoundly in high glucose (P < 0.05 compared to low glucose). Apoptosis was induced as a result of mitochondrial impairment and explained the poor survival of MSCs in high glucose.

Conclusion: High glucose impaired the mitochondrial dynamics and regulatory proteins in hAD-MSCs ensuing their poor survival and high apoptosis rate in hyperglycemic microenvironment.

Keywords: Apoptosis; High glucose; Mesenchymal stem cells; Mitochondrial dynamics; Phosphatidylinositol 3-kinase/mammalian target of rapamycin pathway; Poor survival.