scholarly journals Co-isolation of extracellular vesicles and high-density lipoproteins using density gradient ultracentrifugation

2014 ◽  
Vol 3 (1) ◽  
pp. 23262 ◽  
Author(s):  
Yuana Yuana ◽  
Johannes Levels ◽  
Anita Grootemaat ◽  
Auguste Sturk ◽  
Rienk Nieuwland
Keyword(s):  
Density Gradient ◽  
Extracellular Vesicles ◽  
High Density ◽  
High Density Lipoproteins ◽  
Density Gradient Ultracentrifugation

Visualization of primate high density lipoproteins isolated by density gradient ultracentrifugation

Lipids ◽  
1978 ◽  
Vol 13 (1) ◽  
pp. 88-91 ◽  
Author(s):  
Jerome L. Hojnacki ◽  
Robert J. Nicolosi ◽  
Norma Llansa ◽  
K. C. Hayes
Keyword(s):  
Density Gradient ◽  
High Density ◽  
High Density Lipoproteins ◽  
Density Gradient Ultracentrifugation

Incorporation of radioactive phospholipid into subclasses of high-density lipoproteins

1983 ◽  
Vol 244 (5) ◽  
pp. E513-E516 ◽  
Author(s):  
A. R. Tall ◽  
C. B. Blum ◽  
S. M. Grundy
Keyword(s):  
Density Gradient ◽  
Specific Activity ◽  
High Density ◽  
High Density Lipoproteins ◽  
Equilibrium Density ◽  
Density Gradient Ultracentrifugation ◽  
Hdl Subfractions ◽  
Time Points ◽  
Plasma High Density

The incorporation of orally administered phospholipid into plasma high-density lipoproteins (HDL) was studied in three subjects. Plasma was analyzed by equilibrium density gradient ultracentrifugation, 5, 6, and 8 h after ingestion of 1.1 g [3H-choline, 14C-dilinoleoyl]phosphatidylcholine. At all time points in all subjects, there was a peak of phosphatidylcholine specific activity in fractions of density approximately 1.10-1.13 g/ml, corresponding to the subclass previously designated HDL2a. There was also a more variable, smaller peak of specific activity of phospholipids in HDL2b (1.063-1.100 g/ml) and in fractions of density approximately 1.19 g/ml. In the 1.10-1.13 fraction, 97 and 71%, respectively, of the 3H and 14C radioactivity were in phospholipids. The 3H/14C ratio was similar in phospholipids of HDL subfractions, the d less than 1.07 fraction, and in the administered phospholipid. The results show preferential transfer or exchange or absorbed phosphatidylcholine into specific subclasses of HDL.

scholarly journals Isolation of human salivary extracellular vesicles by iodixanol density gradient ultracentrifugation and their characterizations

2016 ◽  
Vol 5 (1) ◽  
pp. 30829 ◽  
Author(s):  
Kazuya Iwai ◽  
Tamiko Minamisawa ◽  
Kanako Suga ◽  
Yasutomo Yajima ◽  
Kiyotaka Shiba
Keyword(s):  
Density Gradient ◽  
Extracellular Vesicles ◽  
Density Gradient Ultracentrifugation

scholarly journals Comparative Evaluation of Methods for Isolating Small Extracellular Vesicles Derived from Pancreatic Cancer Cells

2021 ◽  
Author(s):  
Jie-Min Wang ◽  
Yong-Jiang Li ◽  
Jun-Yong Wu ◽  
Jia-Xin Cai ◽  
Jing Wen ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cancer Cells ◽  
Density Gradient ◽  
Extracellular Vesicles ◽  
Pancreatic Cancer Cell Line ◽  
Size Exclusion ◽  
Density Gradient Ultracentrifugation ◽  
High Concentration ◽  
Protein Marker ◽  
Pancreatic Cancer Cells

Abstract Background: Small extracellular vesicles (sEVs) are nanosized vesicles involved in cell-to-cell communication. sEVs have been widely studied for clinical applications such as early detection of diseases and as therapeutics. Various methods for sEVs isolation are been using, but different methods may result in different qualities of sEVs and impact downstream analysis and applications. Here, we compared current isolation methods and performed a comparative analysis of sEVs from supernatant of cultured pancreatic cancer cells.Methods: Ultracentrifugation, ultrafiltration and co-precipitation as concentration methods were firstly evaluated for yield, size, morphology and protein level of pellets. Then, isolate sEVs obtained by four different purification methods: size exclusion chromatography, density gradient ultracentrifugation, ultracentrifugation, and immunoaffinity capturing, were analysed and compared.Results: For the concentration process, ultracentrifugation method obtained high quality and high concentration of pellets. For the purification process, immunoaffinity capturing method obtained the purest sEVs with less contaminants, while density gradient ultracentrifugation-based method obtained sEVs with the smallest size. Proteomic analysis revealed distinct protein contents of purified sEVs from different methods. Conclusions: For isolating sEVs derived from supernatant of cultured pancreatic cancer cell line, ultracentrifugation-based method is recommended for concentration of sEVs, density gradient ultracentrifugation-based method may be applied for obtaining purified sEVs with controlled size, immunoaffinity capturing may be suitable for studies requiring sEVs with high purity but may loss subtypes of sEVs without specific protein marker.

scholarly journals Comparative Evaluation of Methods for Isolating Small Extracellular Vesicles Derived from Pancreatic Cells

2020 ◽  
Author(s):  
Jie-Min Wang ◽  
Yong-Jiang Li ◽  
Jun-Yong Wu ◽  
Jia-Xin Cai ◽  
Jing Wen ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cancer Cells ◽  
Density Gradient ◽  
Extracellular Vesicles ◽  
Cell Communication ◽  
Size Exclusion ◽  
Density Gradient Ultracentrifugation ◽  
Pancreatic Cancer Cells ◽  
Co Precipitation ◽  
Therapeutic Study

Abstract Background: Small extracellular vesicles (sEVs) are nanosized vesicles involved in cell-to-cell communication. sEVs have been widely studied for clinical applications such as early detection of diseases and as therapeutics. Various methods for sEVs isolation have been using, but different methods may result in different qualities of sEVs and impact downstream analysis and applications. Here, we compared current isolation methods and performed a comparative analysis of sEVs derived from pancreatic cancer cells.Results: Ultracentrifugation, ultrafiltration and co-precipitation as concentration methods were firstly evaluated for yield, size, morphology and protein level of pellets. Then, isolate sEVs obtained by four different purification methods: size exclusion chromatography, density gradient ultracentrifugation, ultracentrifugation, and immunoaffinity capturing, were analysed and compared. For the concentration process, ultracentrifugation method obtained high quality and concentration pellets. For the purification process, immunoaffinity capturing method obtained the purest sEVs with less contaminants, while density gradient ultracentrifugation-based method obtained sEVs with the smallest size. Proteomic analysis revealed distinct protein contents of purified sEVs. Conclusions: For isolating sEVs derived from pancreatic cancer cells, ultracentrifugation-based method is recommended for concentration of sEVs, density gradient ultracentrifugation-based method may be suitable for isolation of sEVs for therapeutic study, immunoaffinity capturing may be applied for studies exploring sEVs as biomarkers.

scholarly journals Cushioned-Density Gradient Ultracentrifugation (C-DGUC) improves the isolation efficiency of extracellular vesicles

PLoS ONE ◽  
2019 ◽  
Vol 14 (4) ◽  
pp. e0215324 ◽  
Author(s):  
Phat Duong ◽  
Allen Chung ◽  
Laura Bouchareychas ◽  
Robert L. Raffai
Keyword(s):  
Density Gradient ◽  
Extracellular Vesicles ◽  
Density Gradient Ultracentrifugation
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