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Extraction of Extracellular Vesicles from Whole Tissue
Published on: February 7, 2019
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A new integrated method for tissue extracellular vesicle enrichment and proteome profiling
Miaomiao Zhang1,2, Tong Liu2, Zhuokun Du2
1Department of Immunology, Medical College of Qingdao University Qingdao Shandong 266071 PR China aunp_dna@126.com.
RSC Advances
|November 25, 2022
Summary
A new kit-TiO2 method efficiently isolates pure extracellular vesicles (EVs) from tissues. This technique improves EV enrichment for cancer research and biomarker discovery in hepatocellular carcinoma (HCC).
Area of Science:
- Biochemistry
- Cell Biology
- Oncology
Background:
- Extracellular vesicles (EVs) are crucial for cell-to-cell communication in the tumor microenvironment and hold diagnostic/prognostic significance in cancer.
- Current methods for isolating pure EVs from tissues, such as ultracentrifugation, are inefficient, time-consuming, and yield low purity.
- There is an urgent need for rapid, efficient methods to isolate tissue EVs for biological and clinical applications.
Purpose of the Study:
- To develop a novel, rapid, and efficient method for isolating high-purity extracellular vesicles directly from tissue samples.
- To validate the efficacy of the new isolation method compared to existing techniques.
- To investigate the proteomic differences in liver tissue EVs between healthy and hepatocellular carcinoma (HCC) models.
Main Methods:
- Development of a novel isolation approach combining an extraction kit with titanium dioxide (TiO2) microspheres (kit-TiO2).
- EVs were precipitated from tissue fluid and subsequently enriched using TiO2 microspheres via interaction with lipid bilayer phosphate groups.
- Isolation efficiency and purity were assessed using western blot and proteomic analysis.
Main Results:
- The kit-TiO2 method demonstrated improved purity and enrichment efficiency of isolated tissue EVs compared to conventional methods.
- Proteomic analysis identified 1966 protein groups from tissue EVs.
- Comparison of EVs from healthy and HCC-bearing mice revealed 25 upregulated and 75 downregulated protein groups in HCC EVs, including several HCC-relevant proteins.
Conclusions:
- The novel kit-TiO2 approach provides a powerful tool for efficient enrichment of EVs directly from tissues.
- This method enhances EV isolation purity and recovery, facilitating further biological and clinical studies.
- The identified differentially expressed proteins in HCC EVs suggest potential applications in biomarker discovery and drug screening for HCC.
