Objective To develop a
method of isolation and identification of
exosomes of
bone marrow mesenchymal stem cells.
Methods The
mesenchymal stem cells were cultured by whole
bone marrow adherence
method.A new
reagent -
exosomes extraction kit was used to isolate and collecte
exosomes.The
exosomes were identified by elec-tron
microscopy,
particle size detection,
flow cytometry and
Western blot.Results The expression of CD45 on the surface of the third generation
bone marrow mesenchymal stem cells was negative,and CD73 and CD105 were posi-tive;
exosomes derived from
bone marrow mesenchymal stem cells were round or oval,the size is non-
uniform,the diameter is 30~100 nm,have a complete
membrane structure,and containing low-density substances;
Particle size detection particle diameter of the main peak was 61.25 nm, in which the diameter of particles was about 20-200 nm accounted for 72.4%;exosome expressed CD63 and CD81;The expression of CD9 and CD63 from
cell cul-ture supernatants was positive.Conclusions The
exosomes can be collected in the medium of
mesenchymal stem cells.The
exosomes derived from
bone marrow mesenchymal stem cells can be identified by
electron microscopy,
particle size detection,
flow cytometry and
Western blot.