In the present study, we used the SEC method to separately collect different EV fractions from macrophage-like cells, such as large, medium and small EVs and investigated EV size distribution, morphology under TEM, EV markers (CD9, CD63, and two homologs of HSP90-α and β), molecular transfer to cancer cells, and the alteration of viability in the receiver cancer cells. The gene discussed is CD9; the disease is cancer.