Characterization of 3D-Organotypic Culture of Mouse Adipose-Derived Stem Cells
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ABSTRACT:
ORGANISM(S): Mus musculus
PROVIDER: GSE248060 | GEO | 2023/11/22
REPOSITORIES: GEO
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Project description:Although stem cells are a promising avenue for harnessing the potential of adipose tissue, conventional two-dimensional (2D) culture methods have limitations. This study explored the use of three-dimensional (3D) cultures to preserve the regenerative potential of adipose-derived stem cells (ADSCs) and investigated their cellular properties. Flow cytometric analysis revealed significant variations in surface marker expressions between the two culture conditions. While 2D cultures showed robust surface marker expressions, 3D cultures exhibited reduced levels of CD44, CD90.2, and CD105. Adipogenic differentiation in 3D organotypic ADSCs faced challenges, with decreased organoid size and limited activation of adipogenesis-related genes. Key adipocyte markers, such as lipoprotein lipase (LPL) and adipoQ, were undetectable in 3D-cultured ADSCs, unlike positive controls in 2D-cultured mesenchymal stem cells (MSCs). Surprisingly, 3D-cultured ADSCs underwent mesenchymal-epithelial transition (MET), evidenced by increased E-cadherin and EpCAM expression and decreased mesenchymal markers. This study highlights successful ADSC organoid formation, notable MSC phenotype changes in 3D culture, adipogenic differentiation challenges, and a distinctive shift toward an epithelial-like state. These findings offer insights into the potential applications of 3D-cultured ADSCs in regenerative medicine, emphasizing the need for further exploration of underlying molecular mechanisms.
| S-EPMC11011465 | biostudies-literature
Project description:Adipose tissue harbours a significant number of multipotent adult stem cells of mesenchymal origin known as adipose-derived stem cells (ADSCs). Broad differentiation potential and convenient accessibility of ADSCs make them an attractive source of adult mesenchymal stem cell for regenerative medicine and cell developmental plasticity research. Genome-wide microarray expression profiling was performed to identify genes deregulated during osteogenic differentiation of ADSCs to evaluate developmental plasticity of these cells. Dynamics of epigenetic modifications were analyzed in parallel and associated with the gene expression profile. Gene expression profile was analyzed in adipose-derived stem cells (ADSCs) differentiated into osteogenic lineage from 3 donors and compared to undifferentiated cells from the same donors.
2015-01-11 | E-GEOD-63754 | biostudies-arrayexpress
Project description:Adipose tissue harbours a significant number of multipotent adult stem cells of mesenchymal origin known as adipose-derived stem cells (ADSCs). Broad differentiation potential and convenient accessibility of ADSCs make them an attractive source of adult mesenchymal stem cell for regenerative medicine and cell developmental plasticity research. Genome-wide microarray expression profiling was performed to identify genes deregulated during osteogenic differentiation of ADSCs to evaluate developmental plasticity of these cells. Dynamics of epigenetic modifications were analyzed in parallel and associated with the gene expression profile.
2015-01-11 | GSE63754 | GEO
Project description:Characterization of 3D-Organotypic Culture of Mouse Adipose-Derived Stem Cells
| PRJNA1041470 | ENA
Project description:Obesity and type 2 diabetes are strongly associated with adipose tissue dysfunction and impaired adipogenesis. Understanding the molecular underpinnings that control adipogenesis is thus of fundamental importance for the development of novel therapeutics against metabolic disorders. However, translational approaches are hampered as current models do not accurately recapitulate adipogenesis. Here, a scaffold-free versatile 3D adipocyte culture platform with chemically defined conditions is presented in which primary human preadipocytes accurately recapitulate adipogenesis. Following differentiation, multi-omics profiling and functional tests demonstrate that 3D adipocyte cultures feature mature molecular and cellular phenotypes similar to freshly isolated mature adipocytes. Spheroids exhibit physiologically relevant gene expression signatures with 4704 differentially expressed genes compared to conventional 2D cultures (false discovery rate < 0.05), including the concerted expression of factors shaping the adipogenic niche. Furthermore, lipid profiles of >1000 lipid species closely resemble patterns of the corresponding isogenic mature adipocytes in vivo (R2 = 0.97). Integration of multi-omics signatures with analyses of the activity profiles of 503 transcription factors using global promoter motif inference reveals a complex signaling network, involving YAP, Hedgehog, and TGFβ signaling, that links the organotypic microenvironment in 3D culture to the activation and reinforcement of PPARγ and CEBP activity resulting in improved adipogenesis.
| S-EPMC8373086 | biostudies-literature
Project description:Dedifferentiated Fat Cells (DAs) and Adipose-derived stem cells (ADSCs) are two main stem cells derived from adipose tissue. In order to explore whether there is difference in transcriptome between the two cells after multiple subcultures, and which kind of cell should be selected for different regenerative aim, high-throughput RNAseq were conducted to find differentially expressed genes. The subcutaneous adipose tissue of three young women (23-30 years old)patients with normal BMI (19-24) were obtained during liposuction operation on abdomen or thigh. Stromal Vascular Fraction (SVF) and adipocytes were obtained by collagenase digestion, ADSCs was cultured regularly, and DAs were cultured by a modified ceiling culture method. Only 186 genes with different expression between the two groups were obtained.The downregulated gene number of DAs vs. ADSCs was 112 and the upregulated number was 74. Many of the differentially expressed genes are involved in the biological functions such as transcription regulator, protein translation regulation, cytokine interaction and energy metabolism regulation. These data may provide a foundation for further clinical administration of stem cells derived from adipose tissues.
2020-08-07 | GSE141708 | GEO
Project description:Adipose-derived stromal cells (ADSCs) represent a promising stem cell source for tissue engineering and cell based therapy. However, long-term in vitro expansion of ADSCs impede stemness maintaining, partly attributed to deprivation of their original microenvironment. Incompetent cells largely limit the therapeutic effects in ADSCs based clinical use. Therefore, reconstructing a more physiologically and physically relevant niche is an ideal strategy to settle this issue and therefore facilitates their extensive applications in clinical. Here, we transplanted separated ADSCs to local subcutaneous adipose tissues of nude mice as an in vivo cell culture model. We found transplanted ADSCs maintained the primitive morphologies, showed improved proliferation and delayed senescence when compared to cells in conventional incubator. Significantly increased expression of stemness-related markers and multi-lineage differentiation abilities were further observed in in vivo cultured ADSCs. At last, sequencing results revealed different expression genes between ADSCs obtained from in vivo and in vitro conditions mainly located in extracellular matrix and extracellular space, which participated in regulating transcription and protein synthesis. Moreover, we found an Egr1 involved signaling pathway might exert a crucial impact on controlling stemness properties. Our findings might collectively pave the way between the bench and beside in regard of ADSCs based application.
2024-03-20 | GSE261680 | GEO
Project description:Project description: Adipose mesenchymal stem cells (ADSCs) are fast gaining popularity in regenerative medicine for their pluripotency and relative use of collection. Currently, we are addressing the use of ADSCs in otolaryngology, for the treatment of vocal fold (VF) paralysis. VF can result from accidentally sectioning the recurrent laryngeal nerve (RLN) during thyroidectomy. This prevents closure of the upper larynx and promotes food aspiration into the lungs. Volume augmenting materials are sometimes injected in order to bring the paralyzed VF in close proximity to the counter lateral VF. The procedure is called injection medialization laryngoplasty (IML); we have recently developed a rabbit IML model for testing biological reagents in order to improve outcomes. As part of these studies ADSCs were differentiated into chondrocytes and their fate was studied in vitro and in situ. We used mass spectrometry proteomics because microarrays based gene expression platforms do not report actual protein levels. The data show protein levels in controls and chondrogenically differentiating rabbit ADSCs at 0, 14 and 28 days, analyzed using nano-flow liquid chromatography electrospray ionization tandem mass spectrometry (nLC-MS/MS).
2018-06-26 | MSV000082505 | MassIVE
Project description:Recent researches identified the existence of adipose derived stem cells (ADSCs) in adipose tissue. Perivascular ADSCs (PV-ADSCs) locate around vasculatures and can differentiate into vascular lineages. However, the detailed cellular heterogeneity within PV-ADSCs has not been investigated. Therefore, we performed single-cell profiling of subcutaneous (S-) and perivascular (PV-) ADSCs from wild-type and obese mice. After referring to the clustering strategies from other ADSCs’ single-cell data, we provided a more comprehensive picture and trajectory, especially for PV-ADSCs. Both single-cell analysis and in vitro experiments revealed that S-ADSCs from obese mice had impaired abilities of cell migration and proliferation compared to wild-type S-ADSCs. PV-ADSCs have distinctively intrinsic properties. We uncovered 4 subpopulations of PV-ADSCs including Dpp4+, Col4a2+, Clec11a+ and Sult1e1+ cells. Notably, the differentiative function of PV-ADSCs towards vascular lineages was mainly attributed to the existence of Clec11a+ subpopulation, which highly expressed Mgp. The present study provided an integrative view of the ADSCs’ variance from the perspective of origins and obesity.
2022-02-23 | GSE172336 | GEO
Project description:3D Adipose Tissue Culture Links the Organotypic Microenvironment to Improved Adipogenesis
| PRJNA774268 | ENA