Isolation and Differentiation of Stromal Vascular Cells to Beige/Brite Cells

Aune, Ulrike Liisberg; Ruiz, Lauren; Kajimura, Shingo

doi:10.3791/50191-v

Cited by 15 publications

(21 citation statements)

References 15 publications

(16 reference statements)

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“…Finally, antimycin A (Ama, 2.5 lM) was added to inhibit mitochondrial respiration, and the respiration values obtained were subtracted from the previous values. For experiments using mature brown adipocytes, BAT was collected from 30-to 35-week-old control or Mfn2-adKO mice and brown adipocytes were isolated as previously described (Aune et al, 2013). Then, the adipocytes, in the top of the supernatant, were place in the respiratory chamber of the O2K Oxygraph, filled with air-saturated mitochondrial respiration buffer (Kulkarni et al, 2016).…”

Section: High-resolution Respirometry Studiesmentioning

confidence: 99%

Mfn2 is critical for brown adipose tissue thermogenic function

et al. 2017

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Mitochondrial fusion and fission events, collectively known as mitochondrial dynamics, act as quality control mechanisms to ensure mitochondrial function and fine‐tune cellular bioenergetics. Defective mitofusin 2 (Mfn2) expression and enhanced mitochondrial fission in skeletal muscle are hallmarks of insulin‐resistant states. Interestingly, Mfn2 is highly expressed in brown adipose tissue (BAT), yet its role remains unexplored. Using adipose‐specific Mfn2 knockout (Mfn2‐adKO) mice, we demonstrate that Mfn2, but not Mfn1, deficiency in BAT leads to a profound BAT dysfunction, associated with impaired respiratory capacity and a blunted response to adrenergic stimuli. Importantly, Mfn2 directly interacts with perilipin 1, facilitating the interaction between the mitochondria and the lipid droplet in response to adrenergic stimulation. Surprisingly, Mfn2‐adKO mice were protected from high‐fat diet‐induced insulin resistance and hepatic steatosis. Altogether, these results demonstrate that Mfn2 is a mediator of mitochondria to lipid droplet interactions, influencing lipolytic processes and whole‐body energy homeostasis.

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Section: High-resolution Respirometry Studiesmentioning

confidence: 99%

Mfn2 is critical for brown adipose tissue thermogenic function

et al. 2017

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“…The stromal vascular fraction (SVF), containing adipose stem cells/preadipocytes, was isolated from iWAT or eWAT as previously described (Aune et al , 2013; Poret et al , 2021). Briefly, iWAT or eWAT of 8–10 weeks C57BL/6J male mice was collected quickly and chopped into small chunks in ice‐cold PBS.…”

Section: Methodsmentioning

confidence: 99%

Role and mechanism of PVN–sympathetic–adipose circuit in depression and insulin resistance induced by chronic stress

Wang,

Sun,

You

et al. 2023

EMBO Reports

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Chronic stress induces depression and insulin resistance, between which there is a bidirectional relationship. However, the mechanisms underlying this comorbidity remain unclear. White adipose tissue (WAT), innervated by sympathetic nerves, serves as a central node in the interorgan crosstalk through adipokines. Abnormal secretion of adipokines is involved in mood disorders and metabolic morbidities. We describe here a brain–sympathetic nerve–adipose circuit originating in the hypothalamic paraventricular nucleus (PVN) with a role in depression and insulin resistance induced by chronic stress. PVN neurons are labelled after inoculation of pseudorabies virus (PRV) into WAT and are activated under restraint stress. Chemogenetic manipulations suggest a role for the PVN in depression and insulin resistance. Chronic stress increases the sympathetic innervation of WAT and downregulates several antidepressant and insulin‐sensitizing adipokines, including leptin, adiponectin, Angptl4 and Sfrp5. Chronic activation of the PVN has similar effects. β‐adrenergic receptors translate sympathetic tone into an adipose response, inducing downregulation of those adipokines and depressive‐like behaviours and insulin resistance. We finally show that AP‐1 has a role in the regulation of adipokine expression under chronic stress.

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“…White AT was collected from 4‐week to 5‐week‐old Asah1 P361R/P361R and WT mice as previously described 17,18 . Briefly, the mice were euthanized by exposure to isoflurane followed by cervical dislocation.…”

Section: Methodsmentioning

confidence: 99%

“…White AT was collected from 4-week to 5-weekold Asah1 P361R/P361R and WT mice as previously described. 17,18 Briefly, the mice were euthanized by exposure to isoflurane followed by cervical dislocation. Subcutaneous and visceral white AT were collected and placed in sterile PBS with 1% bovine serum albumin (BSA) and transported on ice into a laminar flow hood.…”

Section: Isolation Of Asc From White Atmentioning

confidence: 99%

“…ASC isolated from white AT and fibroblasts established from the ears were used for differentiation into adipocytes as described previously. 17,18 Cells were counted using Trypan Blue (Thermo Scientific) exclusion with the Countess II FL automated cell counter (Life Technologies, Carlsbad, California) and 200 000 ASC or fibroblast cells were seeded into one well of 6-well plate and left to grow for 3-4 days to reach 90% confluency. The cell lines were prepared in quadruplicates (with n = 3 per genotype).…”

Section: Differentiation Of Fibroblasts and Preadipocytes Into Mature...mentioning

confidence: 99%

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Skin inflammation and impaired adipogenesis in a mouse model of acid ceramidase deficiency

Rybová

Kuchař

Sikora

et al. 2022

J of Inher Metab Disea

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Acid ceramidase catalyzes the degradation of ceramide into sphingosine and a free fatty acid. Acid ceramidase deficiency results in lipid accumulation in many tissues and leads to the development of Farber disease (FD). Typical manifestations of classical FD include formation of subcutaneous nodules and joint contractures as well as the development of a hoarse voice. Healthy skin depends on a unique lipid profile to form a barrier that confers protection from pathogens, prevents excessive water loss, and mediates cell-cell communication. Ceramides comprise $50% of total epidermis lipids and regulate cutaneous homeostasis and inflammation. Abnormal skin development including visual skin lesions has been reported in FD patients, but a detailed study of FD skin has not been performed. We conducted a pathophysiological study of the skin in our mouse model of FD. We observed altered lipid composition in FD skin dominated by accumulation of all studied ceramide species and buildup of abnormal storage structures affecting mainly the dermis. A deficiency of acid ceramidase activity also led to the activation of inflammatory IL-6/JAK/signal transducer and activator of transcription 3 and noncanonical NF-κB signaling pathways. Last, we report reduced proliferation of FD mouse fibroblasts and adipose-derived stem/stromal cells (ASC) along with impaired differentiation of ASCs into mature adipocytes.

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Isolation and Differentiation of Stromal Vascular Cells to Beige/Brite Cells

Cited by 15 publications

References 15 publications

Mfn2 is critical for brown adipose tissue thermogenic function

Mfn2 is critical for brown adipose tissue thermogenic function

Role and mechanism of PVN–sympathetic–adipose circuit in depression and insulin resistance induced by chronic stress

Skin inflammation and impaired adipogenesis in a mouse model of acid ceramidase deficiency

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