ABSTRACT
Creation of artificial gametes may provide a universal solution for these patients with no gametes. Stem cell technology may provide a way to obtain fully functional gametes. Retinoic acid [RA] can initiate meiosis. Several studies have demonstrated that RA can promote sperm cells differentiation from mouse embryonic stem cells [mESCs] and other cells from human embryonic stem cells [hESCs]. We sought to determine whether RA could promote differentiation of germ cells from hESCs. hESCs were differentiated as embryoid bodies [EBs] in suspension with all-trans RA [atRA] or without atRA for 0, 1, 3, 5 and 7 days, and then the expression of VASA, SCP3, GDF9 and TEKT1 were compared by real-time PCR. The statistical differences were evaluated by one way ANOVA. The expression of germ cell-specific markers including the gonocyte marker VASA, the meiotic marker SCP3, and post meiotic markers, GDF9 and TEKT1, all increased in the presence and absence of RA as EB differentiation progressed. In addition, the expression of these markers increased an average of 9.3, 6.9, 7.2 and 11.8 fold respectively in the presence of RA, compared to the absence of RA, over 5 days differentiation. Our results indicate that hESCs may have the potential to differentiate to primordial germ cells [PGCs] and early gametes. RA can improve germ cells differentiation from hESCs
Subject(s)
Embryonic Stem Cells , Germ Cells , TretinoinABSTRACT
The debate exists whether or not gonadotropin-releasing hormone (GnRH) analogs used in controlled ovarian hyperstimulation (COH) impair endometrial receptivity.Homeobox A11 (Hoxall),Meis homeobox 1 (Meisl),cadherin 1 (Cdhl),and catenin beta 1 (Ctnnbl) are well known to be involved in successful implantation.In this study,the endometrial expression of Hoxall,Meisl,Cdhl,and Ctnnbl during the peri-implantation period was investigated in an in vitro fertilization (IVF) mouse model by real-time RT-PCR and Western blot to evaluate the relationship between Hoxall,Meisl,Cdhl,and Ctnnbl expression and the impact of the COH on endometrial receptivity.The mimic COH protocols included GnRH agonist plus human menopausal gonadotropin (HMG) (GnRH agonist group),GnRH antagonist plus HMG (GnRH antagonist group),and HMG alone (HMG group).The expression levels of Hoxall,Meisl,Cdhl,and Ctnnbl mRNA and protein were decreased in all of the COH groups.The expression levels of Hoxall and Ctnnbl were the lowest in the GnRH agonist group,and those of Meisl and Cdbl were lower in the GnRH analog groups than the HMG group.There were positive correlations between the expression of Hoxall and Ctnnbl,as well as the expression of Meisl and Cdhl among all the groups.In conclusion,the COH protocols,particularly with GnRH analogs,suppressed Hoxall,Meisl,Ctnnbl and Cdhl expression,in mouse endometrium during the peri-implantation period.Our data reveal a novel molecular mechanism by which the COH protocols might impair endometrial receptivity.
ABSTRACT
In order to observe the effect of Bushenantai recipe on the expression of endometrial leukemia-inhibitory factor (LIF) in mice with embryonic implantation dysfunction (EID), 120 Kun- ruing mice post coition were randomized into three groups: normal control group, model group and traditional Chinese medicine group (TCM group) (n=40 in each group). Uterus was collected on the pregnancy day (Pd) 4, 5, 6 after an intravenous injection of Evan's blue. The endometrium was dyed by Evan's blue and the mean points of response were observed on Pd 5. The expression of LIF mRNA and protein was detected by RT-PCR and immunohistochemistry respectively and analyzed statistically by image system. The results showed that the number of implantation sites in model group was remarkably less than in normal control group and TCM group. There was no significant difference between normal control group and TCM group. The expression of L1F mRNA and protein in model group was delayed. Bushenantai recipe could increase the expression of LIF mRNA and protein in endometria of mice with EID. It was suggested that Bushenantai recipe could improve em- bryo implantation of mice with EID by promoting the endometrial LIF expression and endometrial decidualization.
ABSTRACT
To assess the relationship between pronuclear scoring and day-3 embryo quality and pregnancy outcome and to determine the clinical value of pronuclear stage scoring system in human in vitro fertilization-embryo transfer (WF-ET) program, a pronuclear scoring system was used to score zygotes 16-20 h after insemination during conventional WF or intracytoplasmic sperm injec- tion (ICS1). The embryos were classified into groups Z1, Z2, Z3 and Z4. Comparisons were made of the rates of arrested embryos and excellent embryos on day 3. Comparisons of pregnancy outcome were made only in those patients in whom cohorts of similarly Z-scored embryos were transferred. The results showed that there were less arrested embryos and more excellent embryos on day 3 in groups Z1 and Z2 than those in group Z3 and Z4. More embryos arrested and less excellent embryos developed in group Z4 than group Z3. The clinical pregnancy rates resulting from the transfer of single pronuclear score homologous embryo types were similar among groups Z1, Z2 and Z3. Implanta- tion rates of group Z1 were higher (P<0.05) than that of group Z3. These findings suggests that pro- nuclear scoring can predict developmental ability on day 3 and implantation potential. A evaluation that combines the Z-score and day 3 embryo morphology is useful in the determination of the most viable embryos and the number of embryos for transfer.
ABSTRACT
Summary: In order to compare GnRH agonist with antagonist protocol for the same patient during controlled ovarian stimulation cycles, the in vitro fertilization and embryo transfer (IVF-ET) outcome was retrospectively studied in 81 patients undergoing 105 agonist protocols and 88 antagonist protocols. The results showed that there was no statistically significant difference in duration of ovarian stimulation, number of ampoules, oocytes retrieved, serum estradiol (E2) and progesterone (P) levels,thickness of endometrium, the zygote-and blastocyst-developmcnt rate between GnRH agonist and antagonist protocols (P>0.05). High quality embryo rate was higher in antagonist protocols, but there was no significant difference between two protocols. Implantation rate and clinical pregnant rate were significantly higher in antagonist protocol (15.82% and 30.26%, respectively) than in agonist protocol (5.26% and 10.64% respectively (P<0.05). It was concluded GnRH antagonist protocol probably improved the outcome of pregnancy of older patients with a history of multiple failure of IVF-ET in a GnRH protocol.
ABSTRACT
In order to elucidate the function of homeobox A10 gene (HOXA10) and p57 during decidualization our present study was designed to observe the change of HOXA10 and p57 expression and subcellular localization of HOXA10 in the process of endometrial stromal cell (ESC) differentiation in vitro. Decidualization was induced by 0.5 mmol/L 8-Bromo-cAMP (8-Br-cAMP) together with 1x10(-6) mol/L medroxyprogesterone acetate (MPA). Expression of p57 and HOXA10 was detected by RT-PCR and Western blot after 1-day, 2-day, and 4-day treatment (D1, D2, D4). ESCs cultured in 2%FBS for 1 and 4 d were used as control (C1, C4). The location of HOXA10 was detected by indirect immunofluorescence and HOXA10-GFP transfection. The results are as follows: (1) The expression of HOXA10 decreased progressively during the course of decidualization, and showed significant difference compared to control group C4 after 2-day treatment (D2). (2) On the contrary, the expression of p57 increased progressively and also showed significant difference compared to the control group C4 after 2-day treatment (D2). (3) There was no significant change of HOXA10 and p57 expression after culturing ESCs in 2%FBS for 4 d (C1, C4). (4) HOXA10 located in the nucleus throughout the course. Cytoplasm and nucleus shuttle was not detected in the experiment. Our results suggest that the down-regulation of HOXA10 may contribute to the increase of p57 and that the up-regulation of p57 likely plays an important role in ESC differentiation. Progesterone receptor (PR) pathway may participate in promoting ESCs to exit cell cycle and enter differentiation.