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Cells 2019, 8(2), 80; https://doi.org/10.3390/cells8020080

Metformin Increases Proliferative Activity and Viability of Multipotent Stromal Stem Cells Isolated from Adipose Tissue Derived from Horses with Equine Metabolic Syndrome

1
Department of Experimental Biology, The Faculty of Biology and Animal Science, University of Environmental and Life Sciences, 50375 Wroclaw, Poland
2
Laboratory of Gene Expression, Institute of Biotechnology CAS, Biocev, 25250 Vestec, Czech Republic
3
Laboratory of Growth Regulators, Faculty of Science, Palacky University, 78371 Olomouc, Czech Republic
4
Gene Core BIOCEV, Průmyslová 595, 25250 Vestec, Czech Republic
5
Department of Anthropology and Human Genetics, Faculty of Science, Charles University, 12843 Prague, Czech Republic
6
TATAA Biocenter AB, 41103 Gothenburg, Sweden
7
Faculty of Veterinary Medicine, Equine Clinic-Equine Surgery, Justus-Liebig-University, 35392 Giessen, Germany
*
Author to whom correspondence should be addressed.
Received: 5 December 2018 / Revised: 18 January 2019 / Accepted: 21 January 2019 / Published: 22 January 2019
(This article belongs to the Special Issue Adipose-Derived Stromal/Stem Cells)
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Abstract

In this study, we investigated the influence of metformin (MF) on proliferation and viability of adipose-derived stromal cells isolated from horses (EqASCs). We determined the effect of metformin on cell metabolism in terms of mitochondrial metabolism and oxidative status. Our purpose was to evaluate the metformin effect on cells derived from healthy horses (EqASCHE) and individuals affected by equine metabolic syndrome (EqASCEMS). The cells were treated with 0.5 μM MF for 72 h. The proliferative activity was evaluated based on the measurement of BrdU incorporation during DNA synthesis, as well as population doubling time rate (PDT) and distribution of EqASCs in the cell cycle. The influence of metformin on EqASC viability was determined in relation to apoptosis profile, mitochondrial membrane potential, oxidative stress markers and BAX/BCL-2 mRNA ratio. Further, we were interested in possibility of metformin affecting the Wnt3a signalling pathway and, thus, we determined mRNA and protein level of WNT3A and β-catenin. Finally, using a two-tailed RT-qPCR method, we investigated the expression of miR-16-5p, miR-21-5p, miR-29a-3p, miR-140-3p and miR-145-5p. Obtained results indicate pro-proliferative and anti-apoptotic effects of metformin on EqASCs. In this study, MF significantly improved proliferation of EqASCs, which manifested in increased synthesis of DNA and lowered PDT value. Additionally, metformin improved metabolism and viability of cells, which correlated with higher mitochondrial membrane potential, reduced apoptosis and increased WNT3A/β-catenin expression. Metformin modulates the miRNA expression differently in EqASCHE and EqASCEMS. Metformin may be used as a preconditioning agent which stimulates proliferative activity and viability of EqASCs. View Full-Text
Keywords: adipose-derived stromal cells; equine metabolic syndrome; metformin adipose-derived stromal cells; equine metabolic syndrome; metformin
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Smieszek, A.; Kornicka, K.; Szłapka-Kosarzewska, J.; Androvic, P.; Valihrach, L.; Langerova, L.; Rohlova, E.; Kubista, M.; Marycz, K. Metformin Increases Proliferative Activity and Viability of Multipotent Stromal Stem Cells Isolated from Adipose Tissue Derived from Horses with Equine Metabolic Syndrome. Cells 2019, 8, 80.

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