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 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 cancer cells

2021 ◽  
Vol 11 (1) ◽  
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 Small extracellular vesicles: from mediating cancer cell metastasis to therapeutic value in pancreatic cancer

2022 ◽  
Vol 20 (1) ◽  
Author(s):  
Wenjie Zhang ◽  
Juan Xing ◽  
Tian Liu ◽  
Jie Zhang ◽  
Zhujiang Dai ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Extracellular Matrix ◽  
Extracellular Vesicles ◽  
Cell Communication ◽  
Bioactive Substances ◽  
Early Biomarkers ◽  
Pancreatic Cancer Cells ◽  
Cell Metastasis ◽  
Therapeutic Value ◽  
Distant Organ

AbstractPancreatic cancer is a highly malignant tumor and, is extremely difficult to diagnose and treat. Metastasis is one of the critical steps in the development of cancer and uses cell to cell communication to mediate changes in the microenvironment. Small extracellular vesicles (sEVs)-carry proteins, nucleic acids and other bioactive substances, and are important medium for communication between cells. There are two primary steps in sVEs-mediated metastasis: communication between pancreatic cancer cells and their surrounding microenvironment; and the communication between primary tumor cells and distant organ cells in distant organs that promotes angiogenesis, reshaping extracellular matrix, forming immunosuppressive environment and other ways to form appropriate pre-metastasis niche. Here, we explore the mechanism of localization and metastasis of pancreatic cancer and use sEVs as early biomarkers for the detection and treatment of pancreatic cancer. Graphical Abstract

scholarly journals Stroma-derived extracellular vesicles deliver tumor-suppressive miRNAs to pancreatic cancer cells

Oncotarget ◽  
2017 ◽  
Vol 9 (5) ◽  
pp. 5764-5777 ◽  
Author(s):  
Song Han ◽  
David H. Gonzalo ◽  
Michael Feely ◽  
Carlos Rinaldi ◽  
Sayali Belsare ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cancer Cells ◽  
Extracellular Vesicles ◽  
Pancreatic Cancer Cells ◽  
Tumor Suppressive Mirnas

scholarly journals Hypoxia alters the release and size distribution of extracellular vesicles in pancreatic cancer cells to support their adaptive survival

2019 ◽  
Vol 121 (1) ◽  
pp. 828-839 ◽  
Author(s):  
Mary C. Patton ◽  
Haseeb Zubair ◽  
Mohammad Aslam Khan ◽  
Seema Singh ◽  
Ajay P. Singh
Keyword(s):  
Pancreatic Cancer ◽  
Cancer Cells ◽  
Size Distribution ◽  
Extracellular Vesicles ◽  
Pancreatic Cancer Cells

Extracellular vesicles derived from pancreatic cancer cells BXPC3 or breast cancer cells MCF7 induce a permanent procoagulant shift to endothelial cells

2020 ◽  
Vol 187 ◽  
pp. 170-179 ◽  
Author(s):  
Rania AmraneDjedidi ◽  
Aurélie Rousseau ◽  
Annette K. Larsen ◽  
Ismail Elalamy ◽  
Patrick Van Dreden ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Pancreatic Cancer ◽  
Endothelial Cells ◽  
Cancer Cells ◽  
Extracellular Vesicles ◽  
Breast Cancer Cells ◽  
Pancreatic Cancer Cells

scholarly journals Identification of Dissociation Factors in Pancreatic Cancer using a Mass Spectrometry-Based Proteomic Approach

2019 ◽  
Author(s):  
Peng Liu ◽  
Lingming Kong ◽  
Keke Liang ◽  
Yunhao Wu ◽  
Haoyi Jin ◽  
...  
Keyword(s):  
Pancreatic Cancer ◽  
Cancer Cells ◽  
Size Exclusion ◽  
Serum Free Medium ◽  
Free Medium ◽  
Protein Database ◽  
Serum Free ◽  
The Real ◽  
Pancreatic Cancer Cells ◽  
Medium Group

Abstract Pancreatic cancer is a highly malignant tumor of the digestive system. This secretome of pancreatic cancer is key to its progression and metastasis. In this study, different protein pretreatment methods were used to analyze the secretome, furthermore to identify dissociation factors in pancreatic cancer. Pancreatic cancer cells were cultured in serum-containing or serum-free medium, and the corresponding supernatants were extracted as samples. Subsequently, the above samples were separated by size exclusion chromatography (SEC), and peptide segments were identified by LC-MS/MS. The final results were identified via the hamster secreted protein database and a public database. Although the number of identified proteins in the serum-free medium group was high, the real secretion of proteins in pancreatic cancer cells was changed. There were six significant secreted proteins in the serum-containing medium group. Survival analysis via the TCGA database suggested that patients with higher expression levels of YWHAG showed a worse overall survival rate than those with lower YWHAG expression. YWHAG could be used as a prognostic indicator for pancreatic cancer. Finally, the results in the serum-containing medium group were more similar to the real secretome of pancreatic cancer cells, and the results might be used to identify biomarkers in liquid biopsy.

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