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Culture of mature adipocytes under a permeable membrane and comparative analysis with different cell culture models

ABSTRACT: Culture of mature adipocytes under a permeable membrane and comparative analysis with different cell culture models

PROVIDER: PRJNA473526 | ENA |

REPOSITORIES: ENA

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Culture of mature adipocytes under a permeable membrane and comparative analysis with different cell culture models

Project description:White adipose tissue (WAT) is a central factor in the development of type 2 diabetes. Despite the epidemiological importance of WAT there is a paucity of translational models to study long term changes in mature adipocytes. Here, we describe a novel method for the culture of mature white adipocytes under a permeable membrane. Compared to existing culture methods such as adipose tissue explants and adipocyte ceiling culture, Membrane mature Adipocyte Aggregate Cultures (MAAC) are superior at maintaining adipogenic gene expression through 2 weeks of culture, do not dedifferentiate, and are under reduced hypoxic stress relative to adipose tissue explants. Unbiased RNA-Sequencing analysis indicates that the gene expression profile of MAAC in culture for 1 or 2 weeks is more similar to starting patient material than cultured adipose tissue explants, floating adipocytes, or in vitro-differentiated precursors from the same donor, with the fewest number of differentially expressed genes and the smallest fold-change in gene expression. Importantly, adipocytes from lean and obese patients can be cultured as MAAC, as can subcutaneous and visceral adipocytes, while maintaining depot-specific gene expression signatures. MAAC remain fully functional after long term culture, with similar responses to insulin and lipolytic stimuli compared to recently isolated cells. In addition, MAAC maintain the ability to crosstalk with other cell types, producing synergistic increases in IL6 and IL8 when co-cultured with macrophages, and respond robustly and predictably to diverse pharmacological agonism. Together, these abilities make MAAC a powerful tool for studying phenotypic changes in mature adipocytes, crosstalk between adipocytes and other cell types, and provide an improved translational model for drug development for the modulation of adipose tissue function.

2019-04-02 | GSE115020 | GEO

Scalable production of uniform and mature organoids in a 3D geometrically-engineered permeable membrane

Project description:The application of organoids has been limited by the lack of methods for producing uniformly mature organoids at scale. This study introduces an organoid culture platform, called UniMat, which addresses the challenges of uniformity and maturity simultaneously. UniMat is designed to not only ensure consistent organoid growth but also facilitate an unrestricted supply of soluble factors by a 3D geometrically-engineered, permeable membrane-based platform. Using UniMat, we demonstrate the scalable generation of kidney organoids with enhanced uniformity in both structure and function compared to conventional methods. Notably, kidney organoids within UniMat matured significantly better, showing increased expression of nephron transcripts, more in vivo-like cell-type balance, and better vascularization. Moreover, UniMat's design offers a more standardized organoid model for drug testing, as demonstrated by its consistent response to a polycystic-kidney-disease drug. In essence, UniMat presents a transformative platform for organoid technology, promising applications in organ development, disease modeling, and drug screening.

2024-09-07 | GSE252104 | GEO

Scalable production of uniform and mature organoids in a 3D geometrically-engineered permeable membrane

Project description:Scalable production of uniform and mature organoids in a 3D geometrically-engineered permeable membrane

| PRJNA1057805 | ENA

Examination of mRNA decay rate in mature adipocytes

Project description:Examination of mRNA decay rate in mature adipocytes

| PRJEB34552 | ENA

Pathway-Associated Gene Expression in Gastrointestinal Cell Lines Under Conditions of Culture Age, Dexamethasone Exposure, and miRNA Overexpression

Project description:Adherent cell layers were grown on permeable membrane inserts culture system using multiple GI cell lines.

2019-08-26 | GSE132501 | GEO

Comparative transcriptome analysis of dedifferentiation in porcine mature adipocytes and follicular granulosa cells

Project description:Cellular dedifferentiation signifies the withdrawal of cells from a specific differentiated state into a ‘stem cell’-like undifferentiated state. However, the mechanism of dedifferentiation remains obscure. We showed that mature adipocytes (MA) and follicular granulosa cells (GC), which have distinct functions in vivo, can dedifferentiate during culture in vitro and acquire multipotency. We investigated the dedifferentiation mechanism of MA and GC using global gene expression analyses.

2011-03-10 | GSE17264 | GEO

Examination of mRNA decay rate in mature adipocytes

Project description:The CCR4-NOT deadenylase complex has a critical function to trigger mRNA decay and various biological events. To understand roles of the complex in adipocyte function, we compare mRNA half-lives between control and Cnot complex-deficient mature adipocytes from iWAT and BAT.

2019-10-24 | E-MTAB-8371 | biostudies-arrayexpress

C/EBPα and PPARγ-specific transcriptional programs in mature adipocytes in vivo

Project description:We took a systematic approach to determine the transcriptional programs that are specifically regulated by C/EBPα in mature white adipocytes of mice on chow diet or high fat diet. The hypothesis tested in the present study was that C/EBPα, as a lipogenic transcription factor, has unique direct targets compared to PPARγ. Our inducible adipocyte specific knockout system allows us to test the direct targets of C/EBPα and PPARγ in adipocytes by short-term C/EBPα or PPARγ elimination in mature adipocytes in vivo. Results indicate that although it has been shown that C/EBPα and PPARγ cross-regulate each other, they have distinct direct responsive targets. Moreover, there are very few C/EBPα specific targets in mice on a chow diet, most of the C/EBPα targets in mature adipocytes are genes modulated by HFD feeding.

2014-11-04 | GSE62937 | GEO

C/EBP? and PPAR?-specific transcriptional programs in mature adipocytes in vivo

Project description:We took a systematic approach to determine the transcriptional programs that are specifically regulated by C/EBP? in mature white adipocytes of mice on chow diet or high fat diet. The hypothesis tested in the present study was that C/EBP?, as a lipogenic transcription factor, has unique direct targets compared to PPAR?. Our inducible adipocyte specific knockout system allows us to test the direct targets of C/EBP? and PPAR? in adipocytes by short-term C/EBP? or PPAR? elimination in mature adipocytes in vivo. Results indicate that although it has been shown that C/EBP? and PPAR? cross-regulate each other, they have distinct direct responsive targets. Moreover, there are very few C/EBP? specific targets in mice on a chow diet, most of the C/EBP? targets in mature adipocytes are genes modulated by HFD feeding. Total RNA obtained from subcutaneous adipose tissue of Adn-C/EBP?-/- mice on doxycycline chow diet for 3 days, doxycycline high fat diet for 3 days or 1 month and Adn-PPAR?-/- mice on doxycycline chow diet for 3 days, compared to control littermates.

2014-11-04 | E-GEOD-62937 | biostudies-arrayexpress

Comparative transcriptome analysis of dedifferentiation in mouse mature adipocytes

Project description:Comparative transcriptome analysis of dedifferentiation in mouse mature adipocytes

| PRJNA658115 | ENA