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In vitro model of human subcutaneous adipocyte spheroids for studying mitochondrial dysfunction and mitochondria activating compounds

Title: In vitro model of human subcutaneous adipocyte spheroids for studying mitochondrial dysfunction and mitochondria activating compounds
Authors: Wagner, Anita; Lautaoja-Kivipelto, Juulia H.; Pehkonen, Kalle; Hassinen, Antti; Kuusela, Minna; Röttger, Lisa; Herbers, Elena; Ioannidou, Anna; Mädler, Sophia; Rothenaigner, Ina; Srinivasan, Soumya; Laasonen, Sini; Rahman, M. Tanvir; Elomaa, Pinja; Kortetjärvi, Saija; Olsson, Anneli; Ukkola, Olavi; Hadian, Kamyar; Mann, Matthias; Peltoniemi, Hilkka; Pietiläinen, Kirsi H.; Klingenspor, Martin; Virtanen, Kirsi A.; Hagberg, Carolina E.; Pirinen, Eija
Contributors: CAMM - Research Program for Clinical and Molecular Metabolism; Faculty of Medicine; Institute for Molecular Medicine Finland; HUS Abdominal Center; Department of Medicine; Clinicum; Department of Biochemistry and Developmental Biology; Eija Pirinen / Principal Investigator
Publisher Information: Elsevier Inc.
Publication Year: 2026
Collection: Helsingfors Universitet: HELDA – Helsingin yliopiston digitaalinen arkisto
Subject Terms: Adipocyte; Obesity; Mitochondria; Spheroid; Human metabolism; Lipid; Biomedicine; Genetics; developmental biology; physiology
Description: Mitochondrial abnormalities drive subcutaneous white adipose tissue dysfunction in obesity, yet in vitro models to study adipocyte mitochondria remain limited. Here, we establish a human subcutaneous adipocyte spheroid model to characterize mitochondrial metabolism under obesity-relevant conditions and drug exposure. Human preadipocyte spheroids were differentiated in ultra-low attachment plates for 3 weeks using thiazolidinedione-free medium. Matrigel embedding was incorporated into the protocol as it promoted mitochondrial network and respiration compared to scaffold-free conditions. Differentiated spheroids showed increased lipid accumulation, adipogenic gene expression, mitochondrial respiration, adiponectin secretion, and hormonal responsiveness. Lipid mixture administration during differentiation induced metabolic disturbances, including mitochondrial respiration failure, alongside increased mitochondrial biogenesis. Post-differentiation treatment with rosiglitazone, a peroxisome proliferator-activated receptor γ agonist, improved mitochondrial bioenergetics and adiponectin secretion in lipid mixture-administered adipocyte spheroids. Our model enables precise measurement of adipocyte mitochondria metabolism, providing a platform for mitochondria-focused research and drug discovery in obesity. ; Peer reviewed
Document Type: article in journal/newspaper
File Description: application/pdf
Language: English
Relation: We would like to express our gratitude to Tilkka Hospital, Helsinki, Finland, for their assistance in tissue sample collection and preparation. Imaging was performed at the Biomedicum Imaging Unit, and FIMM High Content Imaging and Analysis unit services of the University of Helsinki, supported by the Helsinki Institute of Life Science (HiLIFE) and Biocenter Finland. These core facilities were supporting imaging and data analysis. We also thank the Chair of Livestock Biotechnology Unit at TUM for their help in freezing spheroids, and the Chair of Nutrition and Immunology at TUM for their assistance with spheroid processing for imaging and cryosectioning. We extend our thanks to Nick Howe from Agilent Technologies for his invaluable help in establishing the Seahorse protocol for adipocyte spheroids. Additionally, we acknowledge Minna Eriksson and Emma Paasikivi for their technical support, Riikka Jokinen for providing materials, and Juha Hulmi for analytical support. This work was supported by the Academy of Finland/Research Council of Finland (335445, 335446, 314455, 314456, 266286, 272376, 314383, and 335443), Finnish Medical Foundation, Finnish Diabetes Research Foundation, Suorsa Foundation, Gyllenberg Foundation, Novo Nordisk Foundation (NNF20OC0060547, NNF17OC0027232, NNF10OC1013354), Paulo Foundation, Sigrid Jusélius Foundation, Government Research Funds, Research Council of Finland Profi6 funding (336449) awarded to the University of Oulu. C.E.H. was supported by Karolinska Institutet (2-1062/2018 and 2-189/2022) and Diabetes Wellness Sweden (DWPG-2022-0032). A.W. was awarded a young investigator start-up project funded by the Deutsche Forschungsgemeinschaft (CRC-TRR333 BATenergy). Open access was funded by Helsinki University Library.; https://hdl.handle.net/10138/627816; 105027545361; 001675340000001
Availability: https://hdl.handle.net/10138/627816
Rights: cc_by ; info:eu-repo/semantics/openAccess ; openAccess
Accession Number: edsbas.A5CDE0A5
Database: BASE