Extracellular vesicles (EV) are lipid-bilayer-capsuled particles released by stress from various cells. It has been recently revealed that EV secretion is altered by a status of cellular senescence. In the present study, we examined the effects of cellular senescence on EV secretion. Neonatal rat cardiac fibroblasts (NRCF)　were treated with doxorubicin (DOX, 24 h) for the induction of cellular senescence. EV were isolated from NRCF culture media using a total exosome isolation kit. The mRNA expression of cell was measured by a real-time quantitative PCR. Protein expression of EV was measured by Western blotting. DOX changed NRCF morphology to swelling one, and increased mRNA expression of p16 and p21, a senescence-associated gene. In EV isolated from NRCF culture media, DOX stimulation increased protein expression of an exosomal maker. Further study is required to elucidate whether senescence-associated EV could affect cellular function as well as protein profiles of their cargo.