Department of Animal Sciences, University of Tennessee, Knoxville, TN, USA; School of Animal Sciences, Virginia Polytechnic Institute and State University, Blacksburg, VA, USA.
Department of Animal Sciences, University of Tennessee, Knoxville, TN, USA.
Anim Reprod Sci. 2023 Jul;254:107264. doi: 10.1016/j.anireprosci.2023.107264. Epub 2023 May 22.
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.
早期牛胚胎由子宫内膜上皮 (EPI) 和基质成纤维细胞 (SF) 细胞分泌的组织滋养分子支持,这些细胞对黄体孕激素 (P4) 有反应。我们假设,特定的组织滋养分子转录丰度取决于细胞类型和 P4 浓度,并且子宫内膜细胞条件培养基 (CM) 可以改善体外生产 (IVP) 的胚胎在培养中的发育。来自 7 个子宫的原代牛 EPI 和 SF 细胞在含有 0(对照)、1、15 或 50ng P4 的 RPMI 培养基中孵育 12 小时。还孵育了不含细胞的 RPMI(N-CM)以及来自 EPI 或 SF 培养物的 CM(EPI-或 SF-CM)或两者的组合(1:1;EPI/SF-CM),用于培养发育第 4-8 天的 IVP 胚胎(n=117)。细胞类型(SLC1A1、SLC5A6、SLC7A1、FGF-2、FGF-7、CTGF、PRSS23 和 NID2)和/或 P4 浓度(FGF-7 和 NID2)对子宫内膜细胞组织滋养分子 mRNA 有影响(P<0.05)。与 N-CM 相比,EPI 或 SF-CM 中第 7 天的囊胚发育更大(P≤0.05),EPI/SF-CM 中囊胚发育更大(P=0.07)。在第 8 天,仅在 EPI-CM 中囊胚发育更大(P<0.05)。此外,用子宫内膜细胞 CM 培养胚胎降低了第 8 天囊胚中细胞黏附分子 LGALS1 的转录丰度(P<0.01)。总之,子宫内膜细胞 CM 或组织滋养分子可用于改善牛的 IVP 胚胎发育。