不同分离技术对滑膜组织培养上清来源外泌体的提取和鉴定

核心信息

  • 论文英文标题: Extraction and identification of synovial tissue-derived exosomes by different separation techniques
  • 期刊: J Orthop Surg Res
  • 发表年份: 2020
  • DOI: 10.1186/s13018-020-01604-x
  • PMID/PMCID: 32151262; PMC7063768

研究概览

The aim of this study is to compare the efficiency of different separation techniques for extracting synovial tissue-derived exosomes. The synovial tissue discarded during knee arthroscopy or total knee arthroplasty surgery was collected from the Third Affiliated Hospital of Beijing University of Chinese Medicine.

核心发现

The extracts of enriched round and discoid vesicles were successfully extracted with UC, SECF, and PEG.

The results of TEM have shown that all three extraction methods can extract circular or elliptical vesicles with disc- and cup-shaped structures from the synovial tissue, with the diameter is about 30-150 nm.

NTA suggested the main peaks of three groups of exosomes are around 100-120 nm, and the concentration of the three groups of exosomes was greater than 1 × 1010/ml.

The results of WB showed that three positive protein markers (CD9, CD63, and Flotillin-1) were highly expressed in the suspension extracted by the three methods and low in the synovial tissue.

研究设计

The synovial tissue discarded during knee arthroscopy or total knee arthroplasty surgery was collected from the Third Affiliated Hospital of Beijing University of Chinese Medicine.

Ultracentrifugation (UC), filtration combined with size exclusion chromatography (SECF), and 8% polyethylene glycol (PEG) were used to extract synovial tissue-derived exosomes.

Transmission electron microscopy (TEM), nanoparticle tracer analysis (NTA), and Western Blot (WB) were used to detect the morphology, particle size, and biomarker proteins (CD9, CD63, Flotillin-1, and calnexin) of exosomes.

Echo Biotech 角色

滑膜组织外泌体分离鉴定

关联平台: Exoomics®, Research Reagents & Tools

关联服务与能力: Tissue EV Isolation & Purification, EV Basic Characterization (TEM/NTA/WB), Research Reagent / Product Supply

关联产品或试剂: Exosupur® EV Isolation/Purification Kit

参考文献

原始论文: Extraction and identification of synovial tissue-derived exosomes by different separation techniques J Orthop Surg Res. 2020. DOI: 10.1186/s13018-020-01604-x. PMID/PMCID: 32151262; PMC7063768.