Extraction and identification of DC2.4 cell-derived exosomes
10.13200/j.cnki.cjb.004432
- VernacularTitle:DC2.4细胞源性外泌体的提取与鉴定
- Author:
Dan LU
- Publication Type:Journal Article
- Keywords:
DC2.4 cells;
Exosomes;
Major histocompatibility complex(MHC)-Ⅰ
- From:
Chinese Journal of Biologicals
2026;39(08):966-970
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo develop a method for extracting DC2.4 cell-derived exosomes, and to analyze the characteristics and identify the functions.MethodsExosomes were extracted from the supernatant of DC2.4 cells by differential centrifugation, and transmission electron microscope was used to observe the morphology of the exosomes.The particle size, polydispersity index(PDI) and Zeta potential were detected by nanoparticle size and Zeta potential analyzer and the expressions of specific markers CD9, TSG101 and Hsp70 were determined by Western blot.The contents of major histocompatibility complex(MHC)-Ⅰ, MHC-Ⅱand CD86 of DC2.4 cell-derived exosomes were detected by ELISA, and the exosomes uptake by RAW264.7 macrophages was observed by fluorescence microscopy.ResultsThe exosomes isolated from the supernatant of DC2.4 cells had a "tea tray" vesicle structure with a diameter of about 30-200 nm.The vesicle size showed a main peak of 154.56 nm and concentrated in(154.56 ± 1.48) nm, PDI was 0.086 ± 0.027, and Zeta potential was(-10.895 ± 1.88)mV.The exosome marker proteins CD9, TSG101 and Hsp70 were expressed.The contents of MHC-Ⅰ, MHC-Ⅱ and CD86 on the surface were 488.94, 762.37 and 0.365 ng/mL, respectively, with significant difference(F=406.2, P<0.000 1).RAW264.7 macrophages were able to uptake DC2.4 cell-derived exosomes.ConclusionA method for extracting DC2.4 cellderived exosomes was successfully established, which lays an experimental foundation for the future exploration on exosomes participating in vaccine delivery.
