Progesterone-stimulated endometrial cell conditioned media increases in vitro produced bovine embryo blastocyst formation

Abstract

The early bovine embryo is supported by histotroph molecules secreted by endometrial epithelial (EPI) and stroma fibroblast (SF) cells in response to luteal progesterone (P4). We hypothesized that specific histotroph molecule transcript abundance depends on cell type and P4 concentration and that endometrial cell conditioned media (CM) could improve in vitro produced (IVP) embryo development in culture. Primary bovine EPI and SF cells from seven uteri were incubated for 12 h with RPMI medium containing 0 (Control), 1, 15, or 50 ng of P4. RPMI was also incubated without cells (N-CM) and CM from EPI or SF cultures (EPI- or SF-CM) or a combination of the two (1:1; EPI/SF-CM) was used to culture IVP embryos from days 4–8 of development (n = 117). There was an effect of cell type (SLC1A1, SLC5A6, SLC7A1, FGF-2, FGF-7, CTGF, PRSS23 and NID2) and/or P4 concentration (FGF-7 and NID2) on endometrial cell histotroph molecule mRNA (P < 0.05). Compared to N-CM, blastocyst development on day 7 was greater in the EPI or SF-CM (P ≤ 0.05) and tended to be greater in the EPI/SF-CM (P = 0.07). On day 8, blastocyst development was greater only in the EPI-CM (P < 0.05). Further, culturing embryos with endometrial cell CM reduced day 8 blastocyst transcript abundance of cell adhesion molecule LGALS1 (P < 0.01). In conclusion, endometrial cell CM or histotroph molecules may be used to improve IVP embryo development in cattle.