Version 1
: Received: 5 October 2023 / Approved: 6 October 2023 / Online: 9 October 2023 (04:32:02 CEST)
Version 2
: Received: 7 November 2023 / Approved: 8 November 2023 / Online: 8 November 2023 (04:07:04 CET)
Kopp, E.L.; Deussen, D.N.; Cuomo, R.; Lorenz, R.; Roth, D.M.; Mahata, S.K.; Patel, H.H. Modeling and Phenotyping Acute and Chronic Type 2 Diabetes Mellitus In Vitro in Rodent Heart and Skeletal Muscle Cells. Cells2023, 12, 2786.
Kopp, E.L.; Deussen, D.N.; Cuomo, R.; Lorenz, R.; Roth, D.M.; Mahata, S.K.; Patel, H.H. Modeling and Phenotyping Acute and Chronic Type 2 Diabetes Mellitus In Vitro in Rodent Heart and Skeletal Muscle Cells. Cells 2023, 12, 2786.
Kopp, E.L.; Deussen, D.N.; Cuomo, R.; Lorenz, R.; Roth, D.M.; Mahata, S.K.; Patel, H.H. Modeling and Phenotyping Acute and Chronic Type 2 Diabetes Mellitus In Vitro in Rodent Heart and Skeletal Muscle Cells. Cells2023, 12, 2786.
Kopp, E.L.; Deussen, D.N.; Cuomo, R.; Lorenz, R.; Roth, D.M.; Mahata, S.K.; Patel, H.H. Modeling and Phenotyping Acute and Chronic Type 2 Diabetes Mellitus In Vitro in Rodent Heart and Skeletal Muscle Cells. Cells 2023, 12, 2786.
Abstract
Type 2 diabetes (T2D) has a complex pathophysiology which makes modeling the disease difficult. We aimed to develop a novel model for simulating T2D in vitro, including hyperglycemia, hyperlipidemia, and variably elevated insulin levels targeting muscle cells. We investigated insulin resistance (IR), cellular respiration, mitochondrial morphometry and the associated function in different T2D -mimicking conditions in rodent skeletal (C2C12) and cardiac (H9C2) myotubes. Physiological controls included 5mM glucose with 20mM mannitol as osmotic controls. To mimic hyperglycemia, cells were exposed to 25mM glucose. Further treatments included insulin, palmitate, or both. After short-term (24h) or long-term (96h) exposure, we performed radioactive glucose uptake and mitochondrial function assays. Mitochondrial size and relative frequencies were assessed by morphometric analyses using electron micrographs. C2C12 and H9C2 cells treated short- or long-term with insulin and/or palmitate and HG showed IR. C2C12 myotubes exposed to T2D -mimicking conditions showed significantly decreased ATP-linked respiration and spare respiratory capacity, and less cytoplasmic area occupied by mitochondria, implying mitochondrial dysfunction. In contrast, H9C2 myotubes showed elevated ATP-linked and maximal respiration and increased cytoplasmic area occupied by mitochondria, indicating better adaptation to stress and compensatory lipid oxidation in a T2D environment. Both cell lines displayed elevated fractions of swollen/vacuolated mitochondria after T2D -mimicking treatments.Our stable and reproducible in vitro model of T2D rapidly induced IR, changes in ATP-linked respiration, shifts in energetic phenotypes, and mitochondrial morphology, which are comparable to the muscles of patients suffering from T2D. Thus, our model should allow studying disease mechanisms, potential new targets, and screen candidate therapeutic compounds.
Keywords
Insulin resistance; type 2 diabetes; cell model; mitochondrial dysfunction
Subject
Medicine and Pharmacology, Endocrinology and Metabolism
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Commenter: Elena Kopp
Commenter's Conflict of Interests: Author