Comparative study of four methods for extracting exosomes derived from human umbilical cord mesenchymal stem cells
10.13200/j.cnki.cjb.004404
- VernacularTitle:4种人脐带间充质干细胞来源外泌体提取方法的比较
- Author:
ZHAO Yongmei
- Publication Type:Journal Article
- Keywords:
Exosome;
Extraction;
Human umbilical cord mesenchymal stem cells(HUC-MSCs)
- From:
Chinese Journal of Biologicals
2025;38(1):89-95
- CountryChina
- Language:Chinese
-
Abstract:
Objective To compare four different methods for extracting exosomes derived from human umbilical cord mesenchymal stem cells(HUCMSCs-Exos) and obtain a simple, efficient, and cost-effective method for isolating exosomes.Methods The(Human umbilical cord Mesenchymal stem cells,HUC-MSCs)were subcultured, observed for cell morphology under microscope, and detected for cell proliferation ability, specific surface antigen expression level and multidirectional differentiation potential. The hucMSCs-Exos were extracted from the culture supernatant of HUC-MSCs by four methods, ultracentrifugation, polyethylene glycol(PEG) precipitation, sucrose precipitation and Total Exosome Isolation kit. The exosomes were observed for the morphology by transmission electron microscope, measured for the concentration and particle size by nanoparticle tracking analysis(NTA), detected for the total protein concentration by BCA, and detected for the specific marker protein level by Western blot.Results The subcultured HUC-MSCs showed long spindle shape and vortex adherence growth under microscope with fast cell proliferation, highly expressed MSCs specific surface antigens CD73 and CD90,and extremely low expressed hematopoietic stem cell surface markers CD34 and CD45, which had the potential to differentiate into adipocytes and osteoblasts. The HUCMSCs-Exos were extracted by all the four methods, except kit method, the exosomes obtained by the other three methods all displayed a cup-shaped double-layer membrane structure with clear boundaries. The total protein concentration and particle concentration of exosomes isolated using the kit were significantly higher than those of the other three methods(F = 35. 18, each P < 0. 01). The exosomes extracted by the four methods all expressed specific marker proteins CD63 and CD81. Except the exosomes extracted by sucrose precipitation did not express CD9, the exosomes extracted by the other methods all expressed CD9.Conclusion The exosomes can be extracted by all the four methods, and the quality of exosomes extracted by ultracentrifugation is stable; the yield of exosomes extracted by the kit is higher, more time-saving and more convenient; the purity of exosomes extracted by PEG precipitation and sucrose precipitation is higher. According to different production requirements, the appropriate exosomes isolation techniques can be selected.
