Production of brown adipocytes

A brown fat cell technology, applied in the field of brown fat cell generation, can solve the problems of insufficient specificity, genetic changes of brown fat cells, and inapplicability to human brown fat tissue

Pending Publication Date: 2020-12-22
SOC DES PROD NESTLE SA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, such techniques have the disadvantage that they may produce genetic alterations that can potentially affect the function of the resulting brown adipocytes
However, this method does not appear to be sufficiently specific nor robust to be used to generate a source of human brown adipose tissue

Method used

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  • Production of brown adipocytes
  • Production of brown adipocytes
  • Production of brown adipocytes

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Experimental program
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Effect test

Embodiment 1

[0227] Materials and methods

[0228] Cell Culture and Differentiation

[0229] Human pluripotent stem cells (PSCs) were harvested at 50,000 cells / cm 2 The human pluripotent stem cells were plated on polystyrene culture plates (Thermo-Fisher, Waltham, MA) coated with Geltrex LDEV-Free hESC qualified low growth factor basement membrane matrix (Thermo-Fisher, Waltham, MA) The dilution was 1:200 in glutamine-free DMEM / F-12 (Corning, Corning, NY). Medium for PSC maintenance, spheroid formation, paraxial mesoderm and brown adipocyte differentiation was achieved using chemically defined basal medium (DBM) supplemented with specific factors. DBM consisted of glutamine-free DMEM / F-12 supplemented with 2% Probumin (EMD Milipore, Billerica, MA), 1x antibiotic-antifungal (Corning, Corning, NY), 1x MEM optional Amino acid (Corning, Corning, NY), 1x trace element A (Corning, Corning, NY), 1x trace element B (Corning, Corning, NY), 1x trace element C (Corning, Corning, NY), 50 μg / mL ...

Embodiment 2

[0254] Example 2 - Human iPSC-BA engraftment in mice

[0255] Human brown adipocytes (hiPSC-derived BA cells) generated by the method of the present invention were implanted into two groups of mice and subcutaneously encapsulated in the device. In one group, mice were injected with forskolin (FSK) to stimulate brown fat cells. In another group, mice implanted with brown fat cells were not stimulated with FSK. Control mice were implanted with empty devices. Three weeks after implantation, all groups were evaluated for PET-SCAN in vivo imaging as well as ex vivo PET-SCAN imaging of the transplanted devices. As analyzed by PET-SCAN ex vivo imaging, the group implanted with FSK-stimulated brown adipocytes showed an increased signal for labeled glucose compared to the group implanted with non-FSK-stimulated brown adipocytes. Mice implanted with empty devices showed no signal.

[0256] Calorimetric analysis was performed on groups of mice implanted with encapsulated human brow...

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Abstract

Use of a transforming growth factor beta (TGF-Beta) signalling inhibitor for producing a population of brown adipocytes in vitro is described.

Description

technical field [0001] The present invention relates to the generation of brown adipocytes. More specifically, the present invention relates to improved methods for differentiating cells, such as human pluripotent stem cells, into brown adipocytes. Background technique [0002] Polyuria (commonly known as diabetes) is a group of metabolic disorders characterized by chronically high blood sugar levels. The global prevalence of diabetes has reached 387 million and is expected to rise to 435 million by 2030. In fact, 65% of the global population lives in countries where obesity causes more deaths than undernutrition (WHO-2010). [0003] There are two main types of diabetes (type 1 and type 2), with different causes and treatments. Type 1 diabetes is caused by destruction of the insulin-secreting beta cells in the pancreas, usually by autoimmune mechanisms. Type 2 diabetes is caused by insulin resistance in peripheral tissues, which can be combined with pancreatic β-cell dys...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): C12N5/077A61K35/35
CPCC12N5/0653C12N2501/999C12N2506/00A61K35/00A61P3/10
Inventor M·克劳泽S·道尔顿J·W·艾弗里三世
Owner SOC DES PROD NESTLE SA
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