Effect of glucose concentration during in vitro culture of mouse embryos on development to blastocyst, success of embryo transfer, and litter sex ratio

The effect of glucose in the medium used during in vitro culture on the sex ratio of bovine blastocysts derived from in-vitro-matured andin-vitro-fertilized oocytes was evaluated. Oocytes were matured and inseminated with mixed sperm from three bulls and were cultured in vitro in modified synthetic oviducal fluid medium with 10% fetal calf serum, with or without glucose supplementation. The overall rate of cleaved embryos that developed to expanded blastocyst in the medium without glucose (27.0%) was significantly greater (P<0.05) than the percentage observed when embryos were cultured in medium with glucose (17.5%). Analysis of variance was performed to analyse the effect of glucose on the proportion of male embryos reaching the blastocyst stage (or arrested at the morula stage) during Days 7 to 10. Regardless of the presence or absence of glucose in the medium, significantly (P<0.05) more male than female embryos were harvested as expanded blastocysts on Day 7 and on Day 8 of culture. On Days 9 plus 10 of culture, a sex ratio imbalance only occurred in the absence of glucose in the culture medium (P<0.05). Glucose did not produce any significant effect on the sex ratio of the overall number of expanded blastocysts harvested by Day 10 of in vitro culture. However a significantly greater proportion of females (P<0.01) were found among those embryos that developed only to the morulae stage after 10 days in vitro. These results show that glucose supplementation of culture media produces a preferential loss of female embryos during culture to the blastocyst stage.

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Faculty Opinions recommendation of In vitro culture of mouse embryos reduces differential gene expression between inner cell mass and trophectoderm.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.14266986.15779098 ◽

2012 ◽

Author(s):

Phyllis Leppert

Keyword(s):

Gene Expression ◽

In Vitro Culture ◽

Differential Gene Expression ◽

Inner Cell Mass ◽

Cell Mass ◽

Mouse Embryos ◽

Differential Gene

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Effects of rate of development of in vitro-produced ovine embryos on sex ratio and in vivo survival after embryo transfer

Theriogenology ◽

10.1016/s0093-691x(97)00378-6 ◽

1997 ◽

Vol 48 (8) ◽

pp. 1369-1378 ◽

Cited By ~ 14

Author(s):

S.L. Catt ◽

J.K. O'Brien ◽

W.M.C. Maxwell ◽

G. Evans

Keyword(s):

Sex Ratio ◽

Embryo Transfer ◽

Rate Of Development

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Effect of glucose concentration and culture substrate on huvec viability in in vitro cultures: A literature review and own results

Journal of Applied Biomedicine ◽

10.1016/j.bbe.2021.04.010 ◽

2021 ◽

Author(s):

Anna Ciechanowska ◽

Ilona Gora ◽

Stanislawa Sabalinska ◽

Piotr Foltynski ◽

Piotr Ladyzynski

Keyword(s):

Literature Review ◽

Glucose Concentration ◽

In Vitro Cultures ◽

Culture Substrate ◽

Effect Of Glucose

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P–218 Analysis of the occurrence of microbial contamination in IVF culture system and the effect of microorganisms on embryo development and clinical outcomes

Human Reproduction ◽

10.1093/humrep/deab130.217 ◽

2021 ◽

Vol 36 (Supplement_1) ◽

Author(s):

F Du ◽

R Li ◽

Q Zhang ◽

W Wang

Keyword(s):

Escherichia Coli ◽

In Vitro Culture ◽

Clinical Outcomes ◽

Embryo Transfer ◽

Follicular Fluid ◽

Culture System ◽

Microbial Contamination ◽

Culture Medium ◽

Ivf Et

Abstract Study question what is the source, prevalence, and influence of microbial contamination on in vitro fertilization (IVF) and embryo transfer (ET) cycles? Summary answer Microbial contamination mainly occurs on Day 2, most caused by Escherichia coli carried with semen. ICSI could prevent contamination effectively and get good clinical outcomes. What is known already Microbial contamination occurs in IVF-ET system occasionally, which is hard to stop happening. The IVF culture system and laboratory environment, the patients’ follicular fluid and semen are not absolutely sterile, while the antibiotics in culture medium isn’t effective for all microbe types, and the artificial operations may bring in microbes. Generally, microbial contamination leads to degradation of embryos, reduction the number of embryos available, and infection of female reproductive tract, which would increase the cost of patients’ time, money, and bring psychological damages. A better understanding of embryo contamination in IVF culture system is of added value. Study design, size, duration A total of 29583 IVF-ET cycles were enrolled in this prospective observational study, from January 2010 to December 2020, included 70 microbial contamination cycles discovered in Day1-Day3 (D1-D3) of in vitro culture. Follicular fluid and semen saved on oocyte retrieval day, and culture medium contaminated were examined and identified for microorganisms at each contamination cycle. Participants/materials, setting, methods Compared the contamination rate of different insemination methods (IVF/ICSI/IVF+ICSI), different in vitro culture days (D1-D3), and different samples examination (follicular fluid, semen, culture medium) respectively, identified the source of microorganism types, compared the IVF culture outcomes and clinical outcomes between total contamination group (TC group, 42 cases) and partial contamination group (PC group, 28 cases). Main results and the role of chance A total of 70 microbial contamination cases occurred in 29583 oocyte retrieving cycles (0.24%), and it was observed only in IVF embryos but never in ICSI (Intracytoplasmic sperm injection) embryos. 38 contamination cases occurred on D2 with a highest ratio (54.3%) compared to D1 (32.9%) and D3(12.9%); Compared with follicular fluid, semen was the main cause inducing contamination from D1 to D3, and Escherichia coli in semen and culture medium, Enterococcus faecalis in follicular fluid proved to be the most common sources. Compared with TC group, the PC group showed a lower rate of No-available embryos (21.4% vs 81.0%) and a higher rate of blastocyst formation (41.2% vs 28.6%), In addition, the clinical pregnancy rate of PC group was higher than that of TC group in both fresh and frozen-thawed embryo transfer cycles (31.3% vs 16.7%, 38.5% vs 0.0%). Limitations, reasons for caution Further study is still necessary to better understand the sources that induce microbial contamination embryos, and more efficient methods are required to remove the microbes on these contaminated embryos so as better develop and manage a sterile micro-environment for successful embryo growth. Wider implications of the findings: The differential embryonic microbe types associated to different IVF culture and clinical outcomes in patients undergoing IVF-ET might have profound implications for understanding the microbial sources and making a better management of IVF culture system. Trial registration number Not applicable

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Diploid and haploid mouse parthenogenetic development following in vitro activation and embryo transfer

Development ◽

10.1242/dev.31.3.635 ◽

1974 ◽

Vol 31 (3) ◽

pp. 635-642

Author(s):

M. H. Kaufman ◽

R. L. Gardner

Keyword(s):

Embryo Transfer ◽

Mouse Embryos ◽

Parthenogenetic Development ◽

In Vitro Activation ◽

Haploid Embryos ◽

Parthenogenetic Mouse Embryos

Parthenogenetic mouse embryos were selected following in vitro activation, and transferred to the oviducts of pseudopregnant recipients. Decidua was evoked by 50–56% of diploid parthenogenones compared to 35·1% of haploid embryos with a single pronucleus, 37·5% of immediate cleavage eggs and 77% of fertilized eggs (controls). On day 4, 58·7% of diploid parthenogenones were morphologically normal morulae or blastocysts; over 90% of these ‘normal’ embryos evoked decidua when retransferred to recipients compared to 8·9% of abnormal embryos flushed from the ‘transfer’ sides, suggesting that only ‘normal’ embryos could evoke decidua. Potentially diploid parthenogenones remained diploid on chromosomal examination on day 4.

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Viability and growth of mouse embryos after in vitro culture and fusion

Development ◽

10.1242/dev.23.3.693 ◽

1970 ◽

Vol 23 (3) ◽

pp. 693-704

Author(s):

Patricia Bowman ◽

Anne McLaren

Keyword(s):

In Vitro Culture ◽

Success Rate ◽

Zona Pellucida ◽

Growth Regulation ◽

Excess Mortality ◽

Blastocyst Stage ◽

Mouse Embryos ◽

Foster Mothers ◽

Mouse Eggs

About 80 % of 8-cell mouse eggs developed to the blastocyst stage in culture, whether the zona pellucida was left intact, or removed with pronase (pre-incubated and dialysed) and the eggs then cultured singly or as fused pairs. When pronase was used without prior incubation and dialysis, the success rate was reduced to 50 %. After transfer to uterine foster-mothers, 20–30 % of apparently normal blastocysts cultured with or without the zona, singly or fused, developed into live foetuses, compared with over 50 % of control blastocysts taken directly from the uterus. Some of the excess mortality of cultured embryos took place before implantation and some soon after. The foetuses derived from cultured blastocysts averaged 0·1 g lighter than those derived from control uterine blastocysts similarly transferred. No differences in the weights of the placentae were observed. Foetal and placental weights were unaffected by whether the eggs had been cultured singly or fused, implying that growth regulation of fused embryos is complete by the 17th day of gestation. The longer the eggs were maintained in culture, the lower was their viability after transfer, and the lighter were the foetuses derived from them.

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Normal adenylate ribonucleotide content in mouse embryos homozygous for the t12 mutation

Development ◽

10.1242/dev.78.1.43 ◽

1983 ◽

Vol 78 (1) ◽

pp. 43-51

Author(s):

Horst Spielmann ◽

Robert P. Erickson

Keyword(s):

In Vitro Culture ◽

Normal Values ◽

Firefly Luciferase ◽

Blastocyst Stage ◽

Mouse Embryos ◽

Luciferase Assay ◽

Preimplantation Mouse Embryos ◽

Preimplantation Mouse

The recently improved firefly luciferase assay was used to determine ATP, ADP or AMP in single preimplantation mouse embryos from crosses yielding lethal t12/t12 embryos. Normal values of the three adenylate ribonucleotides were found in freshly collected 2-cell and 4-cell embryos and during in vitro culture to the blastocyst stage. A decrease in adenylate ribonucleotide content was seen in putative t12/t12 embryos only when they were degenerating.

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PSIV-B-38 Late-Breaking: Effect of Glutamate (Glu) on Developmental Block and the MZT Relative Genes Expression in Mouse embryo during in Vitro Culture

Journal of Animal Science ◽

10.1093/jas/skz258.661 ◽

2019 ◽

Vol 97 (Supplement_3) ◽

pp. 330-331

Author(s):

Yu Liu ◽

An Gang Lou ◽

Shuo Yang ◽

Zhong Shu Li ◽

Nan-Zhu Fang

Keyword(s):

In Vitro Culture ◽

Mrna Expression ◽

Marker Gene ◽

Treated Group ◽

Mouse Embryos ◽

Expression Level ◽

Mrna Expression Level ◽

Significant Difference ◽

Developmental Block

Abstract The risk of developmental block in mammal’s embryos is high during in vitro as compare to in vivo environment because the in vitro embryo-culture systems are suboptimal. During in vitro-culture the balance between ROS production and elimination is disturbed and may lead to 2-cell block in mouse embryos [1]. In the current study, we investigated the effects of Glu as anti-developmental block during IVC on ZGA and MZT on mouse embryos. The mouse embryos were divided into control and different level of Glu treated group. The cleavage rate was determined, the ROS and GSH level was investigated using DCHF-DA and CMF2HC respectively. The mRNA expression level of ZGA marker gene such as Eif-1α, Muerv l, Zscan4d and Hsp70.1 was analyzed among the groups using RT-PCR. The transition rate from 2-cell to 4-cell was significantly higher in 6mmol/L Glu treated group as compare to control and others treated groups. No significant difference was recorded in the level of ROS and GSH during MZT stage among the different groups. The mRNA expression level of ZGA marker gene was significantly increased at middle and late stage in 6mmol/L Glu treated group as compare to control and others treated groups. In conclusion, this study shows that the concentration of 6mmol/L Glu could maintain the dynamic balance of GSH and ROS, increase the expression of ZGA marker gene and maintain its high expression pattern of time series, directly participate in the ZGA activated process; ultimately reduce the risk of developmental block to ensure the successful completion of MZT. Reference [1] Lee MT, Bonneau AR, Giraldez AJ.Zygotic Genome Activation during the Maternal-to-Zygotic Transition. Annual Rev Cell Dev Biol [J], 2014, 30:581–613.

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