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Article

The Metabolic Changes between Monolayer (2D) and Three-Dimensional (3D) Culture Conditions in Human Mesenchymal Stem/Stromal Cells Derived from Adipose Tissue

by
Paulina Rybkowska
1,
Klaudia Radoszkiewicz
1,
Maria Kawalec
2,
Dorota Dymkowska
3,
Barbara Zabłocka
2,
Krzysztof Zabłocki
3 and
Anna Sarnowska
1,*
1
Translational Platform for Regenerative Medicine, Mossakowski Medical Research Institute, Polish Academy of Sciences, 02-106 Warsaw, Poland
2
Molecular Biology Unit, Mossakowski Medical Research Institute, Polish Academy of Sciences, 02-106 Warsaw, Poland
3
Laboratory of Cellular Metabolism, Nencki Institute of Experimental Biology, Polish Academy of Science, 02-093 Warsaw, Poland
*
Author to whom correspondence should be addressed.
Cells 2023, 12(1), 178; https://doi.org/10.3390/cells12010178
Submission received: 23 November 2022 / Revised: 22 December 2022 / Accepted: 30 December 2022 / Published: 1 January 2023
(This article belongs to the Special Issue Stem Cells, Metabolism and Neurodegenerative Diseases)
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Abstract

Introduction: One of the key factors that may influence the therapeutic potential of mesenchymal stem/stromal cells (MSCs) is their metabolism. The switch between mitochondrial respiration and glycolysis can be affected by many factors, including the oxygen concentration and the spatial form of culture. This study compared the metabolic features of adipose-derived mesenchymal stem/stromal cells (ASCs) and dedifferentiated fat cells (DFATs) cultivated as monolayer or spheroid culture under 5% O2 concentration (physiological normoxia) and their impact on MSCs therapeutic abilities. Results: We observed that the cells cultured as spheroids had a slightly lower viability and a reduced proliferation rate but a higher expression of the stemness-related transcriptional factors compared to the cells cultured in monolayer. The three-dimensional culture form increased mtDNA content, oxygen consumption rate (OCR) and extracellular acidification rate (ECAR), especially in DFATs-3D population. The DFATs spheroids also demonstrated increased levels of Complex V proteins and higher rates of ATP production. Moreover, increased reactive oxygen species and lower intracellular lactic acid levels were also found in 3D culture. Conclusion: Our results may suggest that metabolic reconfiguration accompanies the transition from 2D to 3D culture and the processes of both mitochondrial respiration and glycolysis become more active. Intensified metabolism might be associated with the increased demand for energy, which is needed to maintain the expression of pluripotency genes and stemness state.
Keywords: MSC; ASC; DFAT; 3D culture; three-dimensional culture; spheroid; spheres; metabolism; mitochondria; OXPHOS; glycolysis; ATP; hypoxia MSC; ASC; DFAT; 3D culture; three-dimensional culture; spheroid; spheres; metabolism; mitochondria; OXPHOS; glycolysis; ATP; hypoxia

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MDPI and ACS Style

Rybkowska, P.; Radoszkiewicz, K.; Kawalec, M.; Dymkowska, D.; Zabłocka, B.; Zabłocki, K.; Sarnowska, A. The Metabolic Changes between Monolayer (2D) and Three-Dimensional (3D) Culture Conditions in Human Mesenchymal Stem/Stromal Cells Derived from Adipose Tissue. Cells 2023, 12, 178. https://doi.org/10.3390/cells12010178

AMA Style

Rybkowska P, Radoszkiewicz K, Kawalec M, Dymkowska D, Zabłocka B, Zabłocki K, Sarnowska A. The Metabolic Changes between Monolayer (2D) and Three-Dimensional (3D) Culture Conditions in Human Mesenchymal Stem/Stromal Cells Derived from Adipose Tissue. Cells. 2023; 12(1):178. https://doi.org/10.3390/cells12010178

Chicago/Turabian Style

Rybkowska, Paulina, Klaudia Radoszkiewicz, Maria Kawalec, Dorota Dymkowska, Barbara Zabłocka, Krzysztof Zabłocki, and Anna Sarnowska. 2023. "The Metabolic Changes between Monolayer (2D) and Three-Dimensional (3D) Culture Conditions in Human Mesenchymal Stem/Stromal Cells Derived from Adipose Tissue" Cells 12, no. 1: 178. https://doi.org/10.3390/cells12010178

APA Style

Rybkowska, P., Radoszkiewicz, K., Kawalec, M., Dymkowska, D., Zabłocka, B., Zabłocki, K., & Sarnowska, A. (2023). The Metabolic Changes between Monolayer (2D) and Three-Dimensional (3D) Culture Conditions in Human Mesenchymal Stem/Stromal Cells Derived from Adipose Tissue. Cells, 12(1), 178. https://doi.org/10.3390/cells12010178

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