Mature Human White Adipocytes Cultured under Membranes Maintain Identity, Function, and Can Transdifferentiate into Brown-like Adipocytes

Harms, Matthew; Li, Qian; Lee, Sunjae; Zhang, Cheng Cheng; Kull, Bengt; Hallén, Stefan; Thorell, Anders; Alexandersson, Ida; Hagberg, Carolina E.; Peng, Xiaorong; Mardinoğlu, Adil; Spalding, Kirsty L.; Boucher, Jérémie

doi:10.1016/j.celrep.2019.03.026

Cited by 94 publications

(93 citation statements)

References 69 publications

(71 reference statements)

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“…They make it possible to observe regenerating preadipocytes and mesenchymal stem cells [40,41]. The first successful experiments on the cultivation of human adipose tissue explants were published by Smith in 1971, and recently Harms et al published an advancement of the ceiling culture method with mature human adipocytes [42,43]. In recent years, first tissue engineering methods for human adipose tissue have been developed [44,45].…”

Section: Introductionmentioning

confidence: 99%

Examination of ex-vivo viability of human adipose tissue slice culture

et al. 2020

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Obesity is associated with significantly higher mortality rates, and excess adipose tissue is involved in respective pathologies. Here we established a human adipose tissue slice cultures (HATSC) model ex vivo. HATSC match the in vivo cell composition of human adipose tissue with, among others, mature adipocytes, mesenchymal stem cells as well as stroma tissue and immune cells. This is a new method, optimized for live imaging, to study adipose tissue and cell-based mechanisms of obesity in particular. HATSC survival was tested by means of conventional and immunofluorescence histological techniques, functional analyses and live imaging. Surgery-derived tissue was cut with a tissue chopper in 500 μm sections and transferred onto membranes building an air-liquid interface. HATSC were cultured in six-well plates filled with Dulbecco's Modified Eagle's Medium (DMEM), insulin, transferrin, and selenium, both with and without serum. After 0, 1, 7 and 14 days in vitro, slices were fixated and analyzed by morphology and Perilipin A for tissue viability. Immunofluorescent staining against IBA1, CD68 and Ki67 was performed to determine macrophage survival and proliferation. These experiments showed preservation of adipose tissue as well as survival and proliferation of monocytes and stroma tissue for at least 14 days in vitro even in the absence of serum. The physiological capabilities of adipocytes were functionally tested by insulin stimulation and measurement of Phospho-Akt on day 7 and 14 in vitro. Viability was further confirmed by live imaging using Calcein-AM (viable cells) and propidium iodide (apoptosis/necrosis). In conclusion, HATSC have been successfully established by preserving the monovacuolar form of adipocytes and surrounding macrophages and connective tissue. This model allows further analysis of mature human adipose tissue biology ex vivo.

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Section: Introductionmentioning

confidence: 99%

Examination of ex-vivo viability of human adipose tissue slice culture

et al. 2020

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“…Others directly introduced primary adipocytes from mice into mesoscopic, perfused reservoirs 40 . In a recently introduced elegant approach, Harms et al cultured mature human white adipocytes in multiwell plates underneath membranes in a transwell format 41 . All of these innovative approaches, however, did not enable the generation of human white adipose tissue-like structures with in vivo-like physiology.…”

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confidence: 99%

WAT-on-a-chip integrating human mature white adipocytes for mechanistic research and pharmaceutical applications

Rogal

Binder

Kromidas

et al. 2020

Sci Rep

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Obesity and its numerous adverse health consequences have taken on global, pandemic proportions. White adipose tissue (WAt)-a key contributor in many metabolic diseases-contributes about one fourth of a healthy human's body mass. Despite its significance, many WAT-related pathophysiogical mechanisms in humans are still not understood, largely due to the reliance on non-human animal models. In recent years, Organ-on-a-chip (OoC) platforms have developed into promising alternatives for animal models; these systems integrate engineered human tissues into physiological microenvironment supplied by a vasculature-like microfluidic perfusion. Here, we report the development of a novel OoC that integrates functional mature human white adipocytes. The WAT-on-achip is a multilayer device that features tissue chambers tailored specifically for the maintenance of 3D tissues based on human primary adipocytes, with supporting nourishment provided through perfused media channels. The platform's capability to maintain long-term viability and functionality of white adipocytes was confirmed by real-time monitoring of fatty acid uptake, by quantification of metabolite release into the effluent media as well as by an intact responsiveness to a therapeutic compound. The novel system provides a promising tool for wide-ranging applications in mechanistic research of WAtrelated biology, in studying of pathophysiological mechanisms in obesity and diabetes, and in R&D of pharmaceutical industry. The global obesity pandemic poses one of today's biggest challenges to public health. Each year, 2.8 million people die from causes related to overweight or obesity 1. Since the 1970s, the worldwide prevalence of obesity has nearly tripled, also leading to an upsurge in associated comorbidities such as type 2 diabetes (Fig. 1a) 2. Future projections reveal the gravity of this public health crisis: the prevalence rates of childhood obesity are increasing at an alarming pace 3 , and by 2030, more than 50% of U.S. adults are predicted to be obese 4. White adipose tissue (WAT) is the principal organ in obesity. In healthy human adults, WAT comprises approximately 20-25% of the total body mass, thus constituting the second largest organ, after the skin. In obese individuals, WAT's contribution to the total body mass may become as high as 50% (Fig. 1b) 5. WAT is tightly involved in the two most important functions of an organism-energy homeostasis and reproduction 6. In energy homeostasis, not only does WAT act as the main storage site of excess dietary energy (Fig. 1c), it also performs crucial endocrine and metabolic functions (Fig. 1d) 7,8. WAT can sense the body's energy status, and respond appropriately, either by storing fuel, in the form of triacylglycerides, or by releasing it as glycerol and fatty acids, for ultimate delivery to organs in need. While this classically described role of WAT already entailed extensive crosstalk between WAT and other organs, its inter-organ communications extend beyond simple feedback loops activated by fed-or fast...

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“…Earlier studies have shown that the in vitro differentiation of pre adipocytes might be resulting in less mature adipocytes (Volz et al 2019). The maintenance and longterm culture of mature adipocytes has been shown to be successful by others (Harms et al 2019;Louis et al 2019). In the future this in vitro protocol could be carried out with mature adipocytes.…”

Section: Discussionmentioning

confidence: 99%

Development of novel human in vitro vascularized adipose tissue model with functional macrophages

et al. 2020

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Inflammation has been proven significant factor in development of type 2 diabetes. So far, most of the adipose tissue related research has been performed in animals, mainly rodent models. The relevance of translation of animal results to humans is questionable. However, in vitro model with relevant human cell source, such as human adipose tissue stromal cells (hASC), can be developed and should be utilized for human adipose tissue research. We developed in vitro models of human adipose tissue utilizing hASC, endothelial cells and monocytes/macrophages. By isolating endothelial cells and macrophages from same adipose tissue as hASC, we were able to provide method for constructing personalized models of adipose tissue. With these models, we studied the effect of macrophages on adipogenesis and protein secretion, with and without vasculature. The models were analyzed for immunocytochemical markers, cell number, triglyceride accumulation and protein secretion. We found that lipid accumulation was greater in adipocytes in the presence of macrophages. Interferon gamma increased this difference between adipocyte culture and Adipocyte–Macrophage co-culture. Protein secretion was affected more by macrophages when vasculature was not present compared to the mild effect when vasculature was present. The vascularized adipose model with macrophages is valuable tool for human adipose tissue research, especially for the personalized medicine approaches; for choosing the right treatments and for studying rare medical conditions.

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Mature Human White Adipocytes Cultured under Membranes Maintain Identity, Function, and Can Transdifferentiate into Brown-like Adipocytes

Abstract: Highlights d Mature adipocytes cultured as MAAC preserve cellular identity and function d Subcutaneous and visceral adipocytes can be cultured as MAAC d Adipocytes from lean and obese donors can be cultured d Human mature white adipocytes can transdifferentiate into brown-like adipocytes

Cited by 94 publications

References 69 publications

Examination of ex-vivo viability of human adipose tissue slice culture

Examination of ex-vivo viability of human adipose tissue slice culture

WAT-on-a-chip integrating human mature white adipocytes for mechanistic research and pharmaceutical applications

Development of novel human in vitro vascularized adipose tissue model with functional macrophages

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