Project description:We report molecular characterization of human brown and white adipocytes. We showed that PAZ6 and SW872 cells exhibit classical molecular and phenotypic markers of brown and white adipocytes, respectively. However, SGBS cells presented a versatile phenotype of adipocyte Sequencing of three human adipocytes cell lines (SGBS, SW872 and PAZ6) in undifferentiated and differentiated stages.
Project description:We report molecular characterization of human brown and white adipocytes. We showed that PAZ6 and SW872 cells exhibit classical molecular and phenotypic markers of brown and white adipocytes, respectively. However, SGBS cells presented a versatile phenotype of adipocyte
Project description:Interest in human brown fat as a novel therapeutic target to tackle the growing obesity and diabetes epidemic has increased dramatically in recent years. While much insight into brown fat biology has been gained from murine cell lines and models, few resources are available to study human brown fat in-vitro. In this study, we detail the derivation and characterization of a novel human ES UCP1 reporter cell line that marks UCP1 positive adipocytes in-vitro. We targeted a mCherry reporter to the UCP1 stop codon via CRISPR-Cas9 and demonstrated that when differentiated to brown adipocytes, reporter cells express UCP1, display high mitochondrial content and multi-locular lipid morphology, and exhibit functional properties such as lipolysis in the presence of isoproterenol and forskolin. Isolation and purification of mCherry positive cells demonstrated elevated expression of brown fat marker genes and a high similarity to isolated human brown fat versus white fat via RNA-seq. This reporter cell line thus presents new opportunities to study human brown fat biology by enabling future work to understand early human brown fat development, performing disease modeling, and enabling drug screening applications.
Project description:Secreted proteins from adipose tissue play a role in metabolic cross-talk and homeostasis. We performed high sensitivity mass spectrometry-based proteomics on the cell media of human adipocytes derived from the supraclavicular brown adipose and from the subcutaneous white adipose depots of adult humans. We identified 471 potentially secreted proteins covering interesting protein categories such as hormones, growth factors, extracellular matrix proteins and proteins of the complement system, which were differentially regulated in brown and white adipocytes. A total of 101 proteins were exclusively quantified in brown adipocytes and among these were ependymin-related protein 1 (EPDR1). Functional studies suggested a role for EPDR1 in thermogenic adipogenesis. In conclusion, we report substantial differences between the secretomes of brown and white human adipocytes and identify novel candidate batokines that can be important regulators of metabolism.
Project description:Ramirez2017 - Human global metabolism in
brown and white adipocytes
Recon 2.1A, an update to Recon 2.1x, is suitable for
quantitatively-realistic results for flux balance analysis in
human metabolism.
This model is described in the article:
Integrating Extracellular
Flux Measurements and Genome-Scale Modeling Reveals Differences
between Brown and White Adipocytes.
Ramirez AK, Lynes MD, Shamsi F, Xue
R, Tseng YH, Kahn CR, Kasif S, Dreyfuss JM.
Cell Rep 2017 Dec; 21(11):
3040-3048
Abstract:
White adipocytes are specialized for energy storage, whereas
brown adipocytes are specialized for energy expenditure.
Explicating this difference can help identify therapeutic
targets for obesity. A common tool to assess metabolic
differences between such cells is the Seahorse Extracellular
Flux (XF) Analyzer, which measures oxygen consumption and media
acidification in the presence of different substrates and
perturbagens. Here, we integrate the Analyzer's metabolic
profile from human white and brown adipocytes with a
genome-scale metabolic model to predict flux differences across
the metabolic map. Predictions matched experimental data for
the metabolite 4-aminobutyrate, the protein ABAT, and the
fluxes for glucose, glutamine, and palmitate. We also uncovered
a difference in how adipocytes dispose of nitrogenous waste,
with brown adipocytes secreting less ammonia and more urea than
white adipocytes. Thus, the method and software we developed
allow for broader metabolic phenotyping and provide a distinct
approach to uncovering metabolic differences.
This model is hosted on
BioModels Database
and identified by:
MODEL1703310000.
To cite BioModels Database, please use:
Chelliah V et al. BioModels: ten-year
anniversary. Nucl. Acids Res. 2015, 43(Database
issue):D542-8.
To the extent possible under law, all copyright and related or
neighbouring rights to this encoded model have been dedicated to
the public domain worldwide. Please refer to
CC0
Public Domain Dedication for more information.
Project description:In order to select mRNA transcripts strongly enriched in murine white adipocytes versus brown adipocytes or stromal-vascular fraction, gene expression data of the adipocyte and stromal-vascular fractions of the interscapular brown, inguinal subcutaneous as well as visceral epididymal adipose tissue depots of young adult male C57BL/6 mice housed at constant 23°C ambient temperature were obtained. 18 samples: 3 different adipose tissues separated into stromal-vascular fraction and adipocytes, analyzed in biological triplicates.
Project description:In order to select mRNA transcripts strongly enriched in murine white adipocytes versus brown adipocytes or stromal-vascular fraction, gene expression data of the adipocyte and stromal-vascular fractions of the interscapular brown, inguinal subcutaneous as well as visceral epididymal adipose tissue depots of young adult male C57BL/6 mice housed at constant 23°C ambient temperature were obtained.
Project description:Brown adipocytes, muscle and dorsal dermis descend from precursor cells in the dermomyotome, but the factors that regulate commitment to the brown adipose lineage are unknown. Here, we prospectively isolated and determined the molecular profile of embryonic brown preadipose cells. Brown adipogenic precursor activity in embryos was confined to Pdgfrα+, Myf5Cre-lineage-marked cells. RNAseq analysis identified Early B Cell Factor-2 (Ebf2) as one of the most selectively expressed genes in this cell fraction. Importantly, Ebf2-expressing cells purified from Ebf2-GFP embryos or brown fat tissue did not express myoblast or dermal cell markers and uniformly differentiated into brown adipocytes. Interestingly, Ebf2-expressing cells from white fat tissue in adult animals differentiated into brown-like (or beige) adipocytes. Loss of Ebf2 in brown preadipose cells reduced the expression levels of brown preadipose-signature genes, whereas ectopic Ebf2-expression in myoblasts activated brown preadipose-specific genes. Altogether, these results indicate that Ebf2 specifically marks and regulates the molecular profile of brown preadipose cells.
Project description:Brown adipocytes, muscle and dorsal dermis descend from precursor cells in the dermomyotome, but the factors that regulate commitment to the brown adipose lineage are unknown. Here, we prospectively isolated and determined the molecular profile of embryonic brown preadipose cells. Brown adipogenic precursor activity in embryos was confined to Pdgfrα+, Myf5Cre-lineage-marked cells. RNAseq analysis identified Early B Cell Factor-2 (Ebf2) as one of the most selectively expressed genes in this cell fraction. Importantly, Ebf2-expressing cells purified from Ebf2-GFP embryos or brown fat tissue did not express myoblast or dermal cell markers and uniformly differentiated into brown adipocytes. Interestingly, Ebf2-expressing cells from white fat tissue in adult animals differentiated into brown-like (or beige) adipocytes. Loss of Ebf2 in brown preadipose cells reduced the expression levels of brown preadipose-signature genes, whereas ectopic Ebf2-expression in myoblasts activated brown preadipose-specific genes. Altogether, these results indicate that Ebf2 specifically marks and regulates the molecular profile of brown preadipose cells.