scholarly journals Efficiency comparison of B6(Cg)-Tyrc−2j /J and C57BL/6NTac embryos as hosts for the generation of knockout mice

Author(s):  
Yu’e Ma ◽  
Lei He ◽  
Lijie Xiang ◽  
Jie Zhang ◽  
Jing Wang ◽  
...  

AbstractCareful selection of the host embryo is critical to the efficient production of knockout (KO) mice when injecting mouse embryonic stem (mES) cells into blastocysts. B6(Cg)-Tyrc−2j/J (B6 albino) and C57BL/6NTac (B6NTac) strains of mice are widely used to produce host blastocysts for such procedures. Here, we tested these two strains to identify an appropriate match for modified agouti C57BL/6N (JM8A3.N1) mES cells. When comparing blastocyst yield, super-ovulated B6NTac mice produced more injectable blastocysts per female than B6 albino mice (8.2 vs. 5.4). There was no significant difference in birth rate when injected embryos were transferred to the same pseudopregnant recipient strain. However, the live birth rate was significantly higher for B6NTac blastocysts than B6 albino blastocysts (62.7% vs. 50.2%). In addition, the proportion of pups exhibiting high-level and complete chimerism, as identified by coat color, was also significantly higher in the B6NTac strain. There was no obvious difference in the efficiency of germline transmission (GLT) when compared between B6NTac and B6 albino host embryos (61.5% vs. 63.3% for mES clones; 64.5% vs. 67.9% for genes, respectively), thus suggesting that an equivalent GLT rate could be obtained with only a few blastocyst injections for B6NTac embryos. In conclusion, our data indicate that B6NTac blastocysts are a better choice for the microinjection of JM8A3.N1 mES cells than B6 albino blastocysts.

2018 ◽  
Vol 26 (9) ◽  
pp. 1210-1217 ◽  
Author(s):  
Mathilde Bourdon ◽  
Pietro Santulli ◽  
Yulian Chen ◽  
Catherine Patrat ◽  
Khaled Pocate-Cheriet ◽  
...  

Objective: The aim of this study was to assess whether a deferred frozen–thawed embryo transfer (Def-ET) offers any benefits compared to a fresh ET strategy in women who have had 2 or more consecutive in vitro fertilization (IVF)/intracytoplasmic injection (ICSI) cycle failures. Design: An observational cohort study in a tertiary referral care center including 416 cycles from women with a previous history of 2 or more consecutive IVF/ICSI failures cycles. Both Def-ET and fresh ET strategies were compared using univariate and multivariate logistic regression models. The main outcome measured was the cumulative live birth rate (CLBR). Results: A total of 416 cycles were included in the analysis: 197 in the fresh ET group and 219 in the Def-ET group. The CLBR was not significantly different between the fresh and Def-ET groups (58/197 [29.4%] and 57/219 [26.0%], respectively, P = .437). In addition, after the first ET, there was no significant difference in the live birth rate between the fresh ET and Def-ET groups (50/197 [25.4%] vs 44/219 [20.1%], respectively). Multivariate logistic regression analysis indicated that compared to the fresh strategy, the Def-ET strategy was not associated with a higher probability of live birth. Conclusions: In cases with 2 or more consecutive prior IVF/ICSI cycle failures, a Def-ET strategy did not result in better ART outcomes than a fresh ET strategy.


2019 ◽  
Vol 19 (1) ◽  
Author(s):  
Junan Meng ◽  
Mengchen Zhu ◽  
Wenjuan Shen ◽  
Xiaomin Huang ◽  
Haixiang Sun ◽  
...  

Abstract Background It is still uncertain whether surgical evacuation adversely affects subsequent embryo transfer. The present study aims to assess the influence of surgical evacuation on the pregnancy outcomes of subsequent embryo transfer cycle following first trimester miscarriage in an initial in vitro fertilization and embryo transfer (IVF-ET) cycle. Methods A total of 645 patients who underwent their first trimester miscarriage in an initial IVF cycle between January 2013 and May 2016 in Nanjing Drum Tower Hospital were enrolled. Surgical evacuation was performed when the products of conception were retained more than 8 h after medical evacuation. Characteristics and pregnancy outcomes were compared between surgical evacuation patients and no surgical evacuation patients. The pregnancy outcomes following surgical evacuation were further compared between patients with ≥ 8 mm or < 8 mm endometrial thickness (EMT), and with the different EMT changes. Results The EMT in the subsequent embryo transfer cycle of surgical evacuation group was much thinner when compared with that in the no surgical evacuation group (9.0 ± 1.6 mm vs. 9.4 ± 1.9 mm, P = 0.01). There was no significant difference in implantation rate, clinical pregnancy rate, live birth rate or miscarriage rate between surgical evacuation group and no surgical evacuation group (P > 0.05). The live birth rate was higher in EMT ≥ 8 mm group when compared to < 8 mm group in surgical evacuation patients (43.0% vs. 17.4%, P < 0.05). Conclusions There was no significant difference in the pregnancy outcomes of subsequent embryo transfer cycle between surgical evacuation patients and no surgical evacuation patients. Surgical evacuation led to the decrease of EMT, especially when the EMT < 8 mm was association with a lower live birth rate.


2011 ◽  
Vol 105 (02) ◽  
pp. 295-301 ◽  
Author(s):  
Jantien Visser ◽  
Veli-Matti Ulander ◽  
Frans Helmerhorst ◽  
Katja Lampinen ◽  
Laure Morin-Papunen ◽  
...  

SummaryRecurrent miscarriage affects 1–2% of women. In more than half of all recurrent miscarriage the cause still remains uncertain. Thrombophilia has been identified in about 50% of women with recurrent miscarriage and thromboprophylaxis has been suggested as an option of treatment. A randomised double-blind (for aspirin) multicentre trial was performed among 207 women with three or more consecutive first trimester (<13 weeks) miscarriages, two or more second trimester (13–24 weeks) miscarriages or one third trimester fetal loss combined with one first trimester miscarriage. Women were analysed for thrombophilia. After complete work-up, women were randomly allocated before seven weeks’ gestation to either enoxaparin 40 mg and placebo (n=68), enoxaparin 40 mg and aspirin 100 mg (n=63) or aspirin 100 mg (n=76). The primary outcome was live-birth rate. Secondary outcomes were pregnancy complications, neonatal outcome and adverse effects. The 0.92–1.48] was found for enoxaparin and placebo and 65% [RR 1.08, 95% CI 0.83–1.39] for enoxaparin and aspirin when compared to aspirin alone (61%, reference group). In the whole study group the live birth rate was 65% (95% CI 58.66–71.74) for women with three or more miscarriages (n=204). No difference in pregnancy complications, neonatal outcome or adverse effects was observed. No significant difference in live birth rate was found with enoxaparin treatment versus aspirin or a combination of both versus aspirin in women with recurrent miscarriage.


2021 ◽  
Vol 12 ◽  
Author(s):  
Yingying Sun ◽  
Yile Zhang ◽  
Xueshan Ma ◽  
Weitong Jia ◽  
Yingchun Su

BackgroundThe definition of recurrent implantation failure (RIF) differs clinically, one of the most controversial diagnostic criteria is the number of failed treatment cycles. We tried to investigate whether the two implantation failure could be included in the diagnostic criteria of RIF.MethodsA retrospective analysis of the clinical data of patients (N=1518) aged under 40 years with two or more implantation failure, recruited from the Center for Reproductive Medicine of the First Affiliated Hospital of Zhengzhou University from January 2016 to June 2019.ResultsAfter adjusting for confounding factors by using binary logistic regression, the results showed that partial general information and: distribution of associated factors were significant differences such as maternal age (aOR=1.054, P=0.001), type of cycle (aOR=2.040, P&lt;0.001), stage of embryos development (aOR=0.287, P&lt;0.001), number of embryos transferred (aOR=0.184, P&lt;0.001), female factor (tubal pathology) (aOR=0.432, P=0.031) and male factor (aOR=1.734, P=0.002) between the groups with two and three or more unexplained implantation failure. And further explored whether these differential factors had a significant negative impact on pregnancy outcome, the results showed that: for patients who had three unexplained implantation failure, in the fourth cycle of ET, the live birth rate decreased significantly with age (aOR=0.921, P&lt;0.001), and the live birth rate of blastocyst transfer was significantly higher than that of cleavage embryo transfer (aOR=1.826, P=0.007). At their first assisted pregnancy treatment after the diagnosis of RIF according to these two different definitions, there were no significant difference in the biochemical pregnancy rate, clinical pregnancy rate, ectopic pregnancy rate and abortion rate (P&gt;0.05), but the live birth rate (35.64% vs 42.95%, P=0.004) was significantly different. According to the definition of ‘two or more failed treatment cycles’, the live birth rate of the first ET treatment after RIF diagnosis was significantly lower than that of patients according to the definition of ‘three or more failed treatment cycles’.ConclusionFor patients with unexplained recurrent implantation failure, two implantation failure cannot be included in the diagnostic criteria of RIF. This study supports the generally accepted definition of three or more failed treatment cycles for RIF.


2021 ◽  
Vol 12 ◽  
Author(s):  
Jian Xu ◽  
Li Yang ◽  
Zhi-Heng Chen ◽  
Min-Na Yin ◽  
Juan Chen ◽  
...  

ObjectiveTo investigate whether the reproductive outcomes of oocytes with smooth endoplasmic reticulum aggregates (SERa) are impaired.MethodsA total of 2893 intracytoplasmic sperm injection (ICSI) cycles were performed between January 2010 and December 2019 in our center. In 43 transfer cycles, transferred embryos were totally derived from SERa+ oocytes. Each of the 43 cycles was matched with a separate control subject from SERa- patient of the same age ( ± 1 year), embryo condition, main causes of infertility, type of protocols used for fresh or frozen embryo transfer cycles. The clinical pregnancy, implantation, ectopic pregnancy and live birth rate were compared between the two groups.Results43 embryo transfer cycles from SERa- patient were matched to the 43 transferred cycles with pure SERa+ oocytes derived embryos. No significant difference was observed in clinical pregnancy rate (55.81% vs. 65.11%, p=0.5081), implantation rate (47.89% vs. 50.70%, p=0.8667) and live birth rate (48.84% vs. 55.81%, p=0.6659) between the SERa+ oocyte group and the matched group. No congenital birth defects were found in the two groups.ConclusionOur results suggest that the implantation, clinical pregnancy, live birth and birth defects rate of embryos derived from oocytes with SERa are not impaired.


Lupus ◽  
2020 ◽  
Vol 30 (1) ◽  
pp. 70-79
Author(s):  
Ziyi Yang ◽  
Xiangli Shen ◽  
Chuqing Zhou ◽  
Min Wang ◽  
Yi Liu ◽  
...  

Objectives To compare and rank currently available pharmacological interventions for the prevention of recurrent miscarriage (RM) in women with antiphospholipid syndrome (APS). Methods A search was performed using PubMed, Embase, the Cochrane Central Register of Controlled Trials, Web of Science, CNKI, ClinicalTrials.gov, and the UK National Research Register on December 15, 2019. Studies comparing any types of active interventions with placebo/inactive control or another active intervention for the prevention of RM in patients with APS were considered for inclusion. The primary outcomes were efficacy (measured by live birth rate) and acceptability (measured by all-cause discontinuation); secondary outcomes were birthweight, preterm birth, preeclampsia, and intrauterine growth retardation. The protocol of this study was registered with Open Science Framework (DOI: 10.17605/OSF.IO/B9T4E). Results In total, 54 randomized controlled trials (RCTs) comprising 4,957 participants were included. Low-molecular-weight heparin (LMWH) alone, aspirin plus LMWH or unfractionated heparin (UFH), aspirin plus LMWH plus intravenous immunoglobulin (IVIG), aspirin plus LMWH plus IVIG plus prednisone were found to be effective pharmacological interventions for increasing live birth rate (ORs ranging between 2.88 to 11.24). In terms of acceptability, no significant difference was found between treatments. In terms of adverse perinatal outcomes, aspirin alone was associated with a higher risk of preterm birth than aspirin plus LMWH (OR 3.92, 95% CI 1.16 to 16.44) and with lower birthweight than LMWH (SMD −808.76, 95% CI −1596.54 to −5.07). Conclusions Our findings support the use of low-dose aspirin plus heparin as the first-line treatment for prevention of RM in women with APS, and support the efficacy of hydroxychloroquine, IVIG, and prednisone when added to current treatment regimens. More large-scale, high-quality RCTs are needed to confirm these findings, and new pharmacological options should be further evaluated.


2020 ◽  
Author(s):  
Na Li ◽  
Yichun Guan ◽  
Bingnan Ren ◽  
Yuchao Zhang ◽  
Yulin Du ◽  
...  

Abstract Objective To determine whether the morphologic parameters of euploid blastocyst influence the live birth rate (LBR) following single frozen-thawed embryo transfer (FET) cycles? Methods A retrospective cohort analysis involving autologous single FET cycles after next generation sequencing (NGS) based preimplantation genetic testing for aneuploidy (PGT-A) by a large in vitro fertilization (IVF) center that was performed from June 2017 to September 2019.Women were divided into three age groups (< 30, 30–34 and ≥ 35 years old). The primary outcome measure was LBR. Outcomes were compared between different blastocyst quality (Good, Average and Poor), inner cell mass (ICM) grade (A and B), and trophectoderm (TE) grade (A, B and C). Results A total of 232 FET cycles were included, the live birth rate was 48.28%. In the youngest group (< 30 years old, n = 86), LBR were compared between cycles with various blastocyst quality (72.22% for good quality, 54.55% for average quality and 34.78% for poor quality; P = 0.019), ICM grade (70.59% for grade A and 42.03% for grade B; P = 0.035) and TE grade (85.71% for grade A,57.58% for grade B and 34.78 for grade C; P = 0.015). Nevertheless, either in the 30–34 years group (n = 99) or in the oldest group (≥ 35years, n = 47), LBR were also comparable between these subgroups, no significant difference was showed in blastocyst morphologic parameters and LBR (P > 0.05). Furthermore, in the similarly graded euploid blastocysts, there was also no statistical significance in LBR among different age subgroups (P > 0.05). Conclusions In women ≥ 30 years old, euploid blastocyst quality was not associated with the LBR in FET cycles, highlights the development competence of poor-quality euploid blastocysts.


Development ◽  
1994 ◽  
Vol 120 (11) ◽  
pp. 3197-3204 ◽  
Author(s):  
P.A. Labosky ◽  
D.P. Barlow ◽  
B.L. Hogan

Primordial germ cells of the mouse cultured on feeder layers with leukemia inhibitory factor, Steel factor and basic fibroblast growth factor give rise to cells that resemble undifferentiated blastocyst-derived embryonic stem cells. These primordial germ cell-derived embryonic germ cells can be induced to differentiate extensively in culture, form teratocarcinomas when injected into nude mice and contribute to chimeras when injected into host blastocysts. Here, we report the derivation of multiple embryonic germ cell lines from 8.5 days post coitum embryos of C57BL/6 inbred mice. Four independent embryonic germ cell lines with normal male karyotypes have formed chimeras when injected into BALB/c host blastocysts and two of these lines have transmitted coat color markers through the germline. We also show that pluripotent cell lines capable of forming teratocarcinomas and coat color chimeras can be established from primordial germ cells of 8.0 days p.c. embryos and 12.5 days p.c. genital ridges. We have examined the methylation status of the putative imprinting box of the insulin-like growth factor type 2 receptor gene (Igf2r) in these embryonic germ cell lines. No correlation was found between methylation pattern and germline competence. A significant difference was observed between embryonic stem cell and embryonic germ cell lines in their ability to maintain the methylation imprint of the Igf2r gene in culture. This may illustrate a fundamental difference between these two cell types.


2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
F Esiso ◽  
F Lai ◽  
D Cunningham ◽  
D Garcia ◽  
B Barrett ◽  
...  

Abstract Study question Does rapid or delayed insemination after egg retrieval affect fertilization, blastocyst development and live birth rates in CI and ICSI cycles? Summary answer When performing CI or ICSI &lt;1.5h and &gt;6.5h after retrieval, detrimental effects are moderate on fertilization but do not impact blastocyst usage and birth rates. What is known already Several studies have shown that CIor ICSI performed between 3 to 5 h after oocyte retrieval has improved laboratory outcomes. However, some studies indicate that insemination of oocytes, by either CI or ICSI, within 2 hours or more than 8 hours after oocyte retrieval has a detrimental effect on the reproductive outcome. With some ART centres experiencing an increase in workload, respecting these exact time intervals is frequently challenging. Study design, size, duration A single-center retrospective cohort analysis was performed on 6559 patients (9575 retrievals and insemination cycles) between January 1st2017 to July 31st2019. The main outcome measures were live-birth rates. Secondary outcomes included analysis of fertilization per all oocytes retrieved, blastocyst utilization, clinical pregnancy, and miscarriage rates. All analyses used time of insemination categorized in both CI and ICSI cycles. Fertilization rates across categories was analyzed by ANOVA and pregnancy outcomes compared using Chi-square tests. Participants/materials, setting, methods As part of laboratory protocol, oocyte retrieval was performed 36 h post-trigger. Cycles involving injection with testicular/epidydimal sperm, donor or frozen oocytes were excluded. The time interval between oocyte retrieval and insemination was analyzed in eight categories: 0 (0- &lt;0.5h), 1 (0.5-&lt;1.5h), 2 (1.5-&lt;2.5h), 3 (2.5-&lt;3.5h), 4 (3.5-&lt;4.5), 5 (4.5-&lt;5.5), 6 (5.5-&lt;6.5) and 7 (6.5-&lt;8h). The number of retrievals in each group (0–7) was 586, 1594, 1644, 1796, 1836, 1351, 641 and 127 respectively. Main results and the role of chance This study had a mean patient age of 36.0 years and mean of 12.2 oocytes per retrieval in each category. There were 4,955 CI and 4,620 ICSI retrievals. The smallest groups were time category 7 and 0 for CI and ICSI respectively. The results showed that the mean fertilization rate per egg retrieved for CI ranged from 54.1 to 64.9% with a significant difference between time category 0 and 5 (p &lt; 0.001) and category 1 and 5 (p &lt; 0.0.001). Mean fertilization rate for ICSI per egg retrieved ranged from 52.8 to 67.3% with no significant difference between time categories compared to category 5. Blastocyst utilization rate for CI and ICSI were not significantly different for all time categories. In the CI and ICSI groups there were 6,540 and 6,178 total fresh and frozen transfers. The miscarriage and clinical pregnancy rate in CI and ICSI were not significantly different across time categories. The overall mean live birth rate for CI was 32.4% (range: 23.1 to 35.5%). Live-birth rates differed significantly (p = 0.04) in CI with time categories 0 and 7 the lowest. In the ICSI group, the overall mean live birth rate was 30.8% (range: 29.1 to 35.7%),with no significant differences between time categories. Limitations, reasons for caution As this is a retrospective study, the influence of uncontrolled variables cannot be excluded. The group spread was uneven with the early and late time categories having the lowest number of representative retrievals and this could have affected the results obtained. Wider implications of the findings: Our results indicate that both CI and ICSI are optimal when performed between 1.5–6.5 hours after oocyte retrieval. Further prospective studies on reproductive outcomes related to time of insemination are warranted. This data indicates a minimal detrimental effect when it is untenable to follow strict insemination time intervals. Trial registration number 2015P000122


Author(s):  
Ze Wang ◽  
Junli Zhao ◽  
Xiang Ma ◽  
Yun Sun ◽  
Guimin Hao ◽  
...  

Abstract Context Obesity management prior to infertility treatment remains a challenge. To date, results from randomized clinical trials involving weight loss by lifestyle interventions have shown no evidence of improved live birth rate. Objective To determine whether pharmacologic weight-loss intervention before in vitro fertilization and embryo transfer (IVF-ET) can improve live birth rate among overweight or obese women. Design, setting, and participants We conducted a randomized, double-blinded, placebo-controlled trial across 19 reproductive medical centers in China, from July 2017 to January 2019. A total of 877 infertile women scheduled for IVF who had a body mass index of 25kg/m 2 or greater were randomly assigned. Interventions The participants were randomized to receive orlistat (n=439) or placebo (n=438) treatment for 4-12 weeks. Main outcomes and measures Live birth rate after fresh embryo transfer. Results The live birth rate was not significantly different between the two groups (112 of 439 [25.5%] with orlistat and 112 of 438 [25.6%] with placebo; P=.984). No significant differences existed between the groups as to the rates of conception, clinical pregnancy, and pregnancy loss. A statistically significant increase in singleton birthweight was observed after orlistat treatment (3487.50g versus 3285.17g in the placebo group; P=.039). The mean change in body weight during the intervention was −2.49kg in the orlistat group, as compared to −1.22kg in the placebo group, with a significant difference (P=.005). Conclusions Orlistat treatment, prior to IVF-ET, did not improve live birth rate among overweight or obese women, although it was beneficial for weight reduction.


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