ABSTRACT
Background: The short and long co-incubation time of gametes for in vitro fertilization are still debatable issues. This study aims to investigate the effects of short and long co-incubation time of gametes on fertilization, polyspermy, embryonic developmental potential, and clinical outcomes.Methods: Sixty-five patients undergoing IVF treatment were invited to participate in the study between May 2017 and March 2019. Ovarian hyperstimulation was prescribed and oocytes were obtained by trans-vaginal aspiration under ultrasound guidance. Sibling oocytes were randomly allocated to short co-incubation for 4 hours (Group I) in 352 oocytes and long co-incubation for 16-18 hours in 363 oocytes (Group II). Rescue ICSI was carried out if total fertilization failure was documented. Fertilization, embryonic development, and pregnancy outcomes were determined.Results: No significant differences between short and long co-incubation were found in fertilization, polyspermy, cleavage, blastocyst, implantation, clinical pregnancy, and live birth rates.Conclusions: The present study showed that short co-incubation of gametes had no significant difference in fertilization, polyspermy, embryo development, and pregnancy outcomes when compared to long co-incubation. The short co-incubation with early cumulus cell removal and rescue ICSI may have the potential to help a couple who had total fertilization failure.
ABSTRACT
Aim To establish a co-incubation system in cardiac fibroblasts of SD neonatal rats and spleen CD4+ CD25 + regulatory T lymphocytes (Tregs) of normal adult SD rats,and to investigate the effects of eplerenone(EPL) on the interaction of two cells and the relationship with the Kvl.3 channel on Tregs cell membrane.Methods The spleen Tregs of normal adult SD rats were sorted by immunomagnetic bead sorting,and the myocardial fibroblasts of SD rats were isolated by differential adherence method.The experiment was conducted in the following groups:CFs,CFs + Tregs,CFs + Tregs + EPL,Tregs.The proliferation of CFs was detected by CCK-8 method.The expression levels of type Ⅰ collagen,type m collagen and matrix metalloproteinase 2 (MMP-2) secreted by CFs were detected by ELISA.The mRNA expression levels of Kv1.3,KCa3.1 on Tregs cell membrane and intracellular CRAC channel were detected by RT-qPCR technique.Tregs cell membrane Kvl.3 channel protein expression levels were determined by In-Cell Western blot.Results After 48 h incubation of the co-culture system,the cell proliferation was stable.CFs proliferation was marked(P ＜0.01),which could be inhibited by EPL(P ＜0.01).The type Ⅰ,type Ⅲ collagen and MMP-2 secreted by CFs increased (P ＜ 0.01).The expression levels of Kv1.3,KCa3.1 and CRAC channel mRNA in Tregs increased by 6.95,1.99 and 1.53 fold (CFs + Tregs vs Tregs,P ＜0.01),EPL decreased the mRNA level of each channel (CFs +Tregs + EPLvs CFs + Tregs,P＜0.01),and the decrease of Kv1.3 channel was the most significant (P ＜ 0.01).The Kv1.3 channel protein of Tregs increased by 67.9％ (CFs + Tregs vs Tregs,P ＜0.01),which could be inhibited by EPL(P ＜ 0.01).Conclusions Tregs cultured with CFs after 48 h can significantly promote the proliferation of CFs,and EPL can down-regulate the Kv1.3 channel expression on the Tregs membrane and inhibit the activation/proliferation of Tregs,indirectly inhibiting myocardial fibrosis.