Using a 3D Culture System to Differentiate Visceral Adipocytes In Vitro

Emont, Margo P.; Yu, Hui; Jun, Hee-Jin; Hong, Xiangfei; Maganti, Nenita; Stegemann, Jan P.; Wu, Jun

doi:10.1210/en.2015-1567

Cited by 39 publications

(38 citation statements)

References 30 publications

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“…33,34 As observed in 3T3L1 cells, beclin 1 (4.2-fold, p < 0.0001) and FSP27 (3.9-fold, p < 0.001) were upregulated in differentiated SVF, which was associated with significant lipid accumulation and adiponectin expression (Fig. 5).…”

Section: Fsp27 Was Regulated By the Foxo1-autophagy Axis In Adipocytesmentioning

confidence: 52%

“…33,34 Briefly, subcutaneous white adipose tissues from C57BL6/J mice (8-12 week old) were dissected, minced, and digested. Cells were suspended in growth media (DMEM/F12 containing 10% FBS and 100 units/ml penicillin and 100 mg/ml streptomycin (1 £ P/S)), filtered through a 100-micron cell strainer, and centrifuged at 500 x g for 5 minutes.…”

Section: Svf Isolation and Culturementioning

confidence: 99%

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FoxO1 antagonist suppresses autophagy and lipid droplet growth in adipocytes

et al. 2016

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Obesity and related metabolic disorders constitute one of the most pressing heath concerns worldwide. Increased adiposity is linked to autophagy upregulation in adipose tissues. However, it is unknown how autophagy is upregulated and contributes to aberrant adiposity. Here we show a FoxO1-autophagy-FSP27 axis that regulates adipogenesis and lipid droplet (LD) growth in adipocytes. Adipocyte differentiation was associated with upregulation of autophagy and fat specific protein 27 (FSP27), a key regulator of adipocyte maturation and expansion by promoting LD formation and growth. However, FoxO1 specific inhibitor AS1842856 potently suppressed autophagy, FSP27 expression, and adipocyte differentiation. In terminally differentiated adipocytes, AS1842856 significantly reduced FSP27 level and LD size, which was recapitulated by autophagy inhibitors (bafilomycin-A1 and leupeptin, BL). Similarly, AS1842856 and BL dampened autophagy activity and FSP27 expression in explant cultures of white adipose tissue. To our knowledge, this is the first study addressing FoxO1 in the regulation of adipose autophagy, shedding light on the mechanism of increased autophagy and adiposity in obese individuals. Given that adipogenesis and adipocyte expansion contribute to aberrant adiposity, targeting the FoxO1-autophagy-FSP27 axis may lead to new anti-obesity options.

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Section: Fsp27 Was Regulated By the Foxo1-autophagy Axis In Adipocytesmentioning

confidence: 52%

Section: Svf Isolation and Culturementioning

confidence: 99%

FoxO1 antagonist suppresses autophagy and lipid droplet growth in adipocytes

et al. 2016

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“…In addition, Yang et al reported that murine adipose derived stromal cells were successfully differentiated into mature adipocytes with BAT pheno-type and function within 3D hydrogels [50]. For WA, 3D culture has been demonstrated to promote adipogenesis and adipokine secretion compared to 2D culture [51,52]. It has also been demonstrated that a porous structure could improve the cell viability in vitro and promote cell ingrowth and integration in vivo [35,53].…”

Section: Discussionmentioning

confidence: 99%

“…Compared to 2D culture systems, 3D hydrogel based culture systems better retain the characteristics of the depot of origin and promote the adipogenic differentiation of adipogenic progenitor and stromal cells [51,60]. Both HA and gelatin have been widely used as adipose tissue engineering biomaterials [24,61,62].…”

Section: Discussionmentioning

confidence: 99%

Effects of tunable, 3D-bioprinted hydrogels on human brown adipocyte behavior and metabolic function

Kuss

Kim

et al. 2018

Acta Biomaterialia

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Obesity and its related health complications cause billions of dollars in healthcare costs annually in the United States, and there are yet to be safe and long-lasting anti-obesity approaches. Using brown adipose tissue (BAT) is a promising approach, as it uses fats for energy expenditure. However, the effect of the microenvironment on human thermogenic brown adipogenesis and how to generate clinically relevant sized and functioning BAT are still unknown. In our current study, we evaluated the effects of endothelial growth medium exposure on brown adipogenesis of human brown adipose progenitors (BAP). We found that pre-exposing BAP to angiogenic factors promoted brown adipogenic differentiation and metabolic activity. We further 3D bioprinted brown and white adipose progenitors within hydrogel-based bioink with controllable physicochemical properties and evaluated the cell responses in 3D bioprinted environments. We used soft, stiff, and stiff-porous constructs to encapsulate the cells. All three types had high cell viability and allowed for varying levels of function for both white and brown adipocytes. We found that the soft hydrogel constructs promoted white adipogenesis, while the stiff-porous hydrogel constructs improved both white and brown adipogenesis and were the optimal condition for promoting brown adipogenesis. Consistently, stiff-porous hydrogel constructs showed higher metabolic activities than stiff hydrogel constructs, as assessed by 2-deoxy glucose uptake (2-DOG) and oxygen consumption rate (OCR). These findings show that the physicochemical environments affect the brown adipogenesis and metabolic function, and further tuning will be able to optimize their functions. Our results also demonstrate that 3D bioprinting of brown adipose tissues with clinically relevant size and metabolic activity has the potential to be a viable option in the treatment of obesity and type 2 diabetes.

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“…29 Interestingly, VAT stromal cells exhibit additional immunomodulatory properties that may directly or indirectly impact Treg biology, including a heightened secretion of pro-inflammatory cytokines (IL-6 among others) under in vitro conditions, compared with stromal cells isolated from SAT in both humans 69 and mice. 70 Despite recent characterization efforts of VAT and SAT stromal cells at the single-cell level, 27,62,71-73 a more comprehensive analysis using a standardized tissue preparation protocol that yields an accurate representation of all stromal components, as well as consensus markers to render results more comparable, would be very valuable.…”

Section: Depotmentioning

confidence: 99%

Visceral adipose tissue Tregs and the cells that nurture them

Spallanzani

Mathis

2020

Immunological Reviews

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Visceral adipose tissue (VAT) is a primary site for storage of excess energy, but it also serves as an important endocrine organ that impacts organismal metabolism. Chronic, low-grade inflammation of VAT, and eventually systemically, is one of the major drivers of obesity-associated insulin resistance and metabolic abnormalities. A unique population of regulatory T cells (Tregs), with a distinct transcriptional profile and antigen receptor repertoire resides in VAT, keeps inflammation in check and regulates organismal metabolism. Accumulation of these cells depends on interactions with other local immunocytes and, importantly, subtypes of VAT mesenchymal stromal cells (VmSCs) that are either immunomodulators or adipogenic. We summarize our current understanding of the phenotype, function, dependencies, derivation, and modulations of VAT Tregs, and review the heterogeneity and regulation of VmSCs as well as their cross talk with VAT Tregs. Lastly, we discuss imperative questions remaining to be answered. K E Y W O R D Sadipose tissue obesity, IL-33, immunometabolism, mesenchymal stromal cells, Treg cells

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Using a 3D Culture System to Differentiate Visceral Adipocytes In Vitro

Cited by 39 publications

References 30 publications

FoxO1 antagonist suppresses autophagy and lipid droplet growth in adipocytes

FoxO1 antagonist suppresses autophagy and lipid droplet growth in adipocytes

Effects of tunable, 3D-bioprinted hydrogels on human brown adipocyte behavior and metabolic function

Visceral adipose tissue Tregs and the cells that nurture them

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