Bovine uterus-derived exosomes improve developmental competence of somatic cell nuclear transfer embryos

Exosomes widely exist in various tissues and body fluids, including blood, tissue fluid, and urine. In the present study, exosomes were first isolated from the early luteal phase uterus and confirmed through morphological examination, immunofluorescence (IF) staining of special membrane antigen, and Western blot. The effects of exosomes on the developmental competence of somatic cell nuclear transfer (SCNT) embryos were investigated. Transmission electron microscopy results showed that the isolated exsomes were spherical particles with a 50 nm-150 nm diameter. Immunostaining showed that the surface of these isolated particles were CD9 positive, which was confirmed using Western blot. Supplementing SCNT embryos with these isolated exsomes on day 4 of culture significantly increased the blastocyst formation rate (31% vs. 34%, 40.3%, and 34.3%) and hatching rate (30.3% vs. 33.3%, 40.7%, and 35%) in comparison with the non-supplementation (control), and day 3 and day 5 supplementation groups. Blastocysts from the exsome supplementation groups showed higher inner cell mass/trophectoderm cell ratio (48% vs 37.9%) and lower apoptosis index (2.1% vs 6.5%) than the control group. The gene expression analysis of the blastocysts also showed that the exsomes supplementation significantly enhanced the expression levels of IFNT and acrogranin and decreased the expression levels of HSP70, BAX and BIP. In conclusion, the present study indicated that the early luteal phase uterus secretes exosomes, which might play important roles in the development of SCNT embryos.