Straightforward Protocol for Gel-Free Proteomic Analysis of Adipose Tissue

2017 ◽  
pp. 289-296 ◽  
Author(s):  
Yvonne Pasing ◽  
Armin Schniers ◽  
Terkel Hansen
Keyword(s):  
Adipose Tissue ◽  
Proteomic Analysis

Abstract 362: Proteomic Analysis of Low Oxygen Tension-Induced Secretome of Adipose Tissue-Derived Stem Cells

2012 ◽  
Vol 111 (suppl_1) ◽  
Author(s):  
Zeljko Bosnjak ◽  
Bassam Wakim ◽  
Yasheng Yan ◽  
Scott Canfield ◽  
Chika Kikuchi ◽  
...  
Keyword(s):  
Stem Cells ◽  
Adipose Tissue ◽  
Oxygen Tension ◽  
Proteomic Analysis ◽  
Stem Cell Markers ◽  
Paracrine Factors ◽  
Differentiation Potential ◽  
Trophic Effect ◽  
Lower Oxygen

Growing evidence from animal studies shows that adipose tissue-derived stem cells (ASCs) improve cardiac function of infarcted hearts. It is commonly accepted that therapeutic potential of ASCs may depend more on their paracrine effects than differentiation potential. The underlying mechanisms remain unclear. However, most data regarding paracrine factors were obtained from ASCs cultured in normoxic condition (20%). The present study investigated how in vivo physiological oxygen (4%) tension influenced the secretome of ASCs. ASCs were isolated from three 8-week-old BALB/c mice. ASCs were confirmed by the expression of stem cell markers (CD44 and CD90) and their capacity to differentiate into adipocytes and osteocytes. ASCs at passage 5 were cultured in normoxic (20%) and lower oxygen (4%) incubators and conditioned for 24 h (3 cultures/group). The conditioned media (CM) from ASCs were subjected to trypsin digestion followed by analysis using automated nano-flow liquid chromatography tandem mass spectrometry. The collected LC/MS/MS data were searched against the rodent subset of the Uniprot database and the total proteomes were identified. The data were from 6 technical replicates. A total of 28 proteins were identified and 7 proteins were unique to normoxic CM. Of the 21 common proteins detected in both normoxic and lower oxygen CM, 9 were extracellular matrix proteins. The abundance of 6 of these proteins (e.g., collagen I and laminin) differed noticeably between normoxic and lower oxygen CM. In addition, a greater amount of cytokine CXCL5 and matrix metalloproteinase (MMP)-2 was detected in lower oxygen CM than in normoxic CM while tissue inhibitor of metalloproteinase (TIMP)-1 was only detected in normoxic CM. These results indicate that lower oxygen tension differentially regulates the secretome of ASCs. Extrapolating the results of this study to the in vivo setting, it would appear that injected ASCs may exert their anti-fibrotic and trophic effect by 1) directly regulating the balance of MMP/TIMP production and preventing collagen accumulation in ischemic hearts to decrease fibrosis, and 2) secreting trophic factors including CXCL5. These data suggest that proteomic analysis of CM is useful for elucidation of the paracrine effect of ASCs in vivo.

scholarly journals Integrated transcriptomic and proteomic analysis of the molecular cargo of extracellular vesicles derived from porcine adipose tissue-derived mesenchymal stem cells

PLoS ONE ◽  
2017 ◽  
Vol 12 (3) ◽  
pp. e0174303 ◽  
Author(s):  
Alfonso Eirin ◽  
Xiang-Yang Zhu ◽  
Amrutesh S. Puranik ◽  
John R. Woollard ◽  
Hui Tang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Adipose Tissue ◽  
Extracellular Vesicles ◽  
Proteomic Analysis

scholarly journals Proteomic Analysis of Human Brown Adipose Tissue Reveals Utilization of Coupled and Uncoupled Energy Expenditure Pathways

2016 ◽  
Vol 6 (1) ◽  
Author(s):  
Sebastian Müller ◽  
Miroslav Balaz ◽  
Patrik Stefanicka ◽  
Lukas Varga ◽  
Ez-Zoubir Amri ◽  
...  
Keyword(s):  
Adipose Tissue ◽  
Energy Expenditure ◽  
Brown Adipose Tissue ◽  
Proteomic Analysis ◽  
Brown Adipose

scholarly journals Differential Protein Expression in White Adipose Tissue from Obesity-Prone and Obesity-Resistant Mice in Response to High Fat Diet and Anti-Obesity Herbal Medicines

2015 ◽  
Vol 35 (4) ◽  
pp. 1482-1498 ◽  
Author(s):  
Sang Woo Kim ◽  
Tae-Jun Park ◽  
Jae Heon Choi ◽  
Kanikkai Raja Aseer ◽  
Ji-Young Choi ◽  
...  
Keyword(s):  
Body Weight ◽  
Adipose Tissue ◽  
White Adipose Tissue ◽  
Proteomic Analysis ◽  
High Fat Diet ◽  
Body Weight Gain ◽  
Herbal Medicines ◽  
High Fat ◽  
Body Weight Reduction ◽  
Peptide Mass

Background: One of the most interesting issues in obesity research is why certain humans are obesity-prone (OP) while others are obesity-resistant (OR) upon exposure to a high-calorie diet. However, the pathways responsible for these phenotypic differences are still largely unknown. Methods: In order to discover marker molecules determining susceptibility and/or resistance to obesity in response to high fat diet (HFD) or anti-obesity herbal medicine (TH), we conducted comparative proteomic analysis of white adipose tissue (WAT) from OP, OR, as well as TH-treated mice. Results: OP mice fed HFD gained approximately 33% more body weight than OR mice, and TH significantly reduced body weight gain in HFD-fed mice by 30%. These mice were further subjected to proteomic analysis using two-dimensional electrophoresis (2-DE) combined with matrix-assisted laser desorption/ionization-time of flight mass spectrometry (MALDI-TOF-MS). Proteomic data revealed 59 spots that were differentially regulated from a total of 1,045 matched spots, and 57 spots of these were identified as altered WAT proteins between OP and OR mice by peptide mass finger printing. Interestingly, 45 proteins were similarly regulated in OR mice in response to TH treatment. Of these, 10 proteins have already been recognized in the context of obesity; however, other proteins involved in obesity susceptibility or resistance were identified for the first time in the present study. Conclusion: Our results suggest that TH actively contributed to body weight reduction in HFD-fed obese mice by altering protein regulation in WAT, and it was also found that TH-responsive proteins can be used as potent molecules for obesity treatment.

OR5,4 Depot-specific proteomic analysis of adipose tissue from GHR-/- mice

2008 ◽  
Vol 18 ◽  
pp. S11
Author(s):  
L. Sackmann-Sala ◽  
D.E. Berryman ◽  
J.J. Kopchick
Keyword(s):  
Adipose Tissue ◽  
Proteomic Analysis

scholarly journals Comparative proteomic analysis of osteogenic differentiated human adipose tissue and bone marrow‐derived stromal cells

2020 ◽  
Vol 24 (20) ◽  
pp. 11814-11827
Author(s):  
Mehran Dadras ◽  
Caroline May ◽  
Johannes Maximilian Wagner ◽  
Christoph Wallner ◽  
Mustafa Becerikli ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Adipose Tissue ◽  
Stromal Cells ◽  
Proteomic Analysis ◽  
Human Adipose Tissue ◽  
Comparative Proteomic Analysis

Proteomic analysis of adipose tissue: informing diabetes research

2014 ◽  
Vol 11 (4) ◽  
pp. 491-502 ◽  
Author(s):  
Mora Murri ◽  
María Insenser ◽  
Manuel Luque ◽  
Francisco J Tinahones ◽  
Héctor F Escobar-Morreale
Keyword(s):  
Adipose Tissue ◽  
Proteomic Analysis ◽  
Diabetes Research
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