Peptidomic analysis of blastocyst culture medium and the effect of peptide derived from blastocyst culture medium on blastocyst formation and viability

2019 ◽  
Vol 87 (1) ◽  
pp. 191-201
Author(s):  
Hui Ji ◽  
Xiaodan Shi ◽  
Jiayi Wang ◽  
Shanren Cao ◽  
Xiufeng Ling ◽  
...  
Keyword(s):  
Culture Medium ◽  
Blastocyst Formation ◽  
Blastocyst Culture ◽  
Peptidomic Analysis

scholarly journals The impact of selected embryo culture conditions on ART treatment cycle outcomes: a UK national study

2020 ◽  
Vol 2020 (1) ◽  
Author(s):  
Catherine M Castillo ◽  
Joyce Harper ◽  
Stephen A Roberts ◽  
Helen C O’Neill ◽  
Edward D Johnstone ◽  
...  
Keyword(s):  
Embryo Culture ◽  
Culture Medium ◽  
Culture Media ◽  
Wald Test ◽  
Culture Conditions ◽  
Oxygen Level ◽  
Success Rates ◽  
Blastocyst Culture ◽  
Clinic Site ◽  
Medium Type

Abstract STUDY QUESTION Are selected embryo culture conditions namely media, oxygen level, and incubator type, associated with IVF live birth rate (LBR) and the health of singleton offspring at birth? SUMMARY ANSWER There were statistically significant differences in LBR between the eight culture media systems analysed; however, none of the embryo culture factors showed statistically significant associations with birth weight (BW) in multivariable regression analyses. WHAT IS KNOWN ALREADY In clinical ART culture media is the initial environment provided for the growth of human embryos. Pre-implantation development is a critical period of developmental plasticity, which could have long-lasting effects on offspring growth and health. Although some studies have shown an impact of culture medium type on BW, the interaction between culture medium type and associated culture conditions on both treatment success rates (LBR) and offspring BW is largely unexplored. This study aimed to examine these factors in a large multicentre national survey capturing the range of clinical practice. STUDY DESIGN, SIZE, DURATION In this cross-sectional study, data from a survey circulated to all UK IVF clinics requesting information regarding culture medium type, incubator type, and oxygen level used in ART between January 2011 and December 2013 were merged with routinely recorded treatment and outcome data held in the Human Fertilisation and Embryology Authority Register up to the end of 2014. PARTICIPANTS/MATERIALS, SETTING, METHODS Forty-six (62%) UK clinics responded to the survey. A total of 75 287 fresh IVF/ICSI cycles were captured, including 18 693 singleton live births. IVF success (live birth, singleton or multiple; LB), singleton gestation and singleton gestation-adjusted BW were analysed using logistic and linear regression models adjusting for patient/treatment characteristics and clinic-specific effects. MAIN RESULTS AND THE ROLE OF CHANCE Culture medium type was shown to have some impact on LBR (multivariable logistic regression, (MRL); post-regression Wald test, P < 0.001), but not on BW (MLR; post-regression Wald test, P = 0.215). However, blastocyst culture had the largest observed effect on odds of LBR (odds ratio (OR) = 1.35, CI: 1.29–1.42), increased the risk of pre-term birth even when controlling for oxygen tension (MLR; OR = 1.42, CI: 1.23–1.63), and gestation-adjusted BW (MLR, β = 38.97 g, CI: 19.42–58.53 g) when compared to cleavage-stage embryo culture. We noted a very strong effect of clinic site on both LBR and BW, thus confounding between treatment practices and clinic site may have masked the effect of culture conditions. LIMITATIONS, REASONS FOR CAUTION Larger datasets with more inter-centre variation are also needed, with key embryo culture variables comprehensively recorded in national treatment registries. WIDER IMPLICATIONS OF THE FINDINGS This study is the largest investigation of laboratory environmental effects in IVF on both LBR and singleton BW. Our findings largely agree with the literature, which has failed to show a consistent advantage of one culture media type over another. However, we noted some association of LBR with medium type, and the duration of embryo exposure to laboratory conditions (blastocyst culture) was associated with both LBR and singleton health at birth. Because of the strong effect of clinic site noted, further randomized controlled trials are needed in order to reliably determine the effect of embryo culture on IVF success rates and the growth and health of subsequent offspring. STUDY FUNDING/COMPETING INTEREST(S) This study was funded by the EU FP7 project grant EpiHealthNet (FP7-PEOPLE-2012-ITN -317 146). The authors have no competing interests to declare.

67 Effects of phytohemagglutinin on the culture of isolated bovine blastomeres derived from the 8-cell stage invitro-produced embryos

2020 ◽  
Vol 32 (2) ◽  
pp. 159
Author(s):  
Y. Hashiyada ◽  
Y. Aikawa ◽  
H. Matsuda ◽  
T. Yamanouchi
Keyword(s):  
Tissue Culture ◽  
Culture Medium ◽  
Flat Surface ◽  
Formation Rate ◽  
Cell Number ◽  
The Other ◽  
Blastocyst Formation ◽  
Culture Dish ◽  
Cell Stage ◽  
Tissue Culture Dish

Monozygotic twin embryos which can efficiently be produced by blastomere separation and aggregation of early cleavage stages of embryos using commercially provided well-of-the-well (WOW) culture dish. Phytohaemagglutinin (PHA) is a plant lectin that binds to and aggregates on the surface of animal cells, but also contains toxicity that causes food poisoning. The present study was conducted to evaluate the toxicity to embryos and the effect to development of isolated blastomeres on PHA-supplemented WOW culture. Embryos were produced using oocytes from ovaries collected at an abattoir by IVM, IVF, and invitro culture (IVC). The tissue culture medium 199 supplemented with 5% calf serum (CS), Brackett-Oliphant solution supplemented with 10mgmL−1 bovine serum albumin, and CR1aa medium containing 5% CS were used for each culture step. For the evaluation of PHA toxicity, 89 embryos that developed to the 5-8-cell stage were obtained at Day 2 after insemination. Each embryo was cultured in a droplet of 5 µL/embryo IVC culture medium supplemented with or without PHA. For the evaluation of PHA to development of isolated blastomeres, 111 of 8-cell stage embryos were obtained 48-54h post-insemination. Zonae pellucidae were removed by exposure to 0.25% pronase. Then, embryos were separated into single blastomeres by gentle pipetting in IVC medium. Each four blastomeres were formed in the shape of a bunch inside the thin cylinder at the tip of the Pasteur pipette by gentle pipetting. Then, each mass of blastomeres in each 60 masses was cultured individually in 5-µL droplets of IVC medium supplemented with or without PHA on the flat surface of a tissue culture dish. On the other hand, each four blastomeres were introduced into a single conical micro-well each having a diameter and depth of ~287µm and 168µm (Dai Nippon Printing). This culture of blastomeres was performed covered with a droplet of 2.5µL well−1 IVC medium supplemented with or without PHA in each 50 or 52 wells. In all of investigations, PHA was used at 50µgmL−1 (Akagi et al. 2011 J. Reprod. Dev. 57). Statistical analysis was performed using Student's t-test and analysis of variance. The blastocyst formation rate (71.1±2.3% vs. 72.7±1.7%), total cell number (120 vs. 122), and inner cell mass cell number (47 vs. 51) at Day 7 after IVF did not differ between PHA-supplemented and PHA-free group in the toxicity test, respectively. In the blastomere culture, the blastocyst formation rate was very low (10.0±5.9% vs. 5.0±2.9%) regardless of the PHA supplementation in drops on the flat surface of a tissue culture dish. On the other hand, blastocyst formation was improved using the WOW culture dish (24.0±3.6% vs. 40.4±7.6%) but there was no difference with or without PHA supplementation. Although nontoxicity of PHA and efficacy of WOW culture for isolated-aggregated blastomeres were confirmed, no improvement of PHA supplementation on development was observed in this study. Subsequently, experiments on the optimum concentration of PHA for aggregation and development of blastomeres in WOW culture are required.

scholarly journals Hyperglycosylated human chorionic gonadotropin in blastocyst culture medium of male and female human embryos

2019 ◽  
Vol 112 (3) ◽  
pp. e126 ◽  
Author(s):  
Georgi Stamenov Stamenov ◽  
Kristina Nikolova ◽  
Magdalena Vasileva ◽  
Ivaylo Rangelov ◽  
Rumiana Ganeva ◽  
...  
Keyword(s):  
Chorionic Gonadotropin ◽  
Human Chorionic Gonadotropin ◽  
Culture Medium ◽  
Human Embryos ◽  
Blastocyst Culture ◽  
Male And Female ◽  
Hyperglycosylated Human Chorionic Gonadotropin

Minimally invasive preimplantation genetic testing using blastocyst culture medium

2019 ◽  
Vol 34 (7) ◽  
pp. 1369-1379 ◽  
Author(s):  
Jiao Jiao ◽  
Bei Shi ◽  
Matthew Sagnelli ◽  
Dalei Yang ◽  
Yaxin Yao ◽  
...  
Keyword(s):  
Genetic Testing ◽  
Minimally Invasive ◽  
Research And Development ◽  
Culture Medium ◽  
Inner Cell Mass ◽  
Cell Mass ◽  
Development Program ◽  
Blastocyst Culture ◽  
Preimplantation Genetic Testing ◽  
Time Reduction

Abstract STUDY QUESTION Is minimally invasive chromosome screening (MICS) using blastocyst culture medium (BCM) sufficiently fast and accurate for preimplantation genetic testing (PGT) SUMMARY ANSWER A new assay for MICS, named MICS-Inst achieved high-resolution, comprehensive chromosome ploidy detection using BCM. WHAT IS KNOWN ALREADY BCM is a viable source of genomic DNA for use in PGT. STUDY DESIGN, SIZE, DURATION Forty-one vitrified blastocysts donated by 22 couples known to carry a chromosome rearrangement and 21 vitrified blastocysts donated from 8 couples with normal karyotypes were used in this study. Good-quality blastocysts, defined as Day 5 and Day 6 embryos ≥ BB (AA, AB, BA, BB) based on the Gardner system were used for analysis. Recruitment took place from May 2018 to August 2018. We performed PGT for structural rearrangements (PGT-SR) on 41 BCM, trophectoderm (TE) biopsy and blastocyst-stage embryo (BE) samples as well as PGT for aneuploidies (PGT-A) on 21 BCM, TE biopsy and BE samples. PARTICIPANTS/MATERIALS, SETTING, METHODS We made several significant modifications to the BCM composition (mixing blastocoel fluid and spent blastocyst medium) as well as the pre-existing multiple annealing and looping-based amplification cycles (MALBAC) techniques and library generation procedures. The design of a quasilinear preamplification (Pre-AMP) primer and AMP primers 1 and 2 enables the preparation of a next-generation sequencing library after the exponential amplification stage by introducing the Illumina P5 and P7 primers into the final products, which are then ready for sequencing. Sequencing was performed on the Illumina Hiseq 2500 platform with 2.0 Mb raw reads generated for each sample. MAIN RESULTS AND THE ROLE OF CHANCE For PGT-A, BCM and TE biopsy samples showed 90% and 86% clinical concordance with the corresponding BE samples, respectively. In addition, both BCM and TE biopsy samples showed 76% karyotype concordance with the corresponding BE samples. For PGT-SR, we successfully obtained ploidy information for all 23 chromosomes with the exception of any rearrangements involving the Y chromosome. Both BCM and TE biopsy samples showed 100% clinical concordance with the corresponding BE samples in detecting chromosomal rearrangements. BCM and TE biopsy samples showed 90% and 100% karyotype concordance with the corresponding BE samples, respectively. Additionally, no statistically significant differences were detected in the aforementioned values of the BCM and TE biopsy samples in either PGT-A or PGT-SR (P > 0.05). Moreover, we achieved accurate quantification of segmental abnormalities using BCM samples. In addition, MICS-Inst reduced the number of steps required for library preparation through the use of new primer designs, resulting in an overall time reduction of 7.5 h. This time reduction allows for the performance of fresh blastocyst transfers. LIMITATIONS, REASONS FOR CAUTION The main limitation is that BE, rather the inner cell mass, was used as the standard to evaluate the chromosome screening results. WIDER IMPLICATIONS OF THE FINDINGS These results show that MICS-Inst is effective in procedure and precision for PGT, and that it is possible to achieve fresh blastocyst transfer following PGT. The implications are significant, as these findings may lead to minimally invasive PGT methods in the future. STUDY FUNDING/COMPETING INTEREST(S) This work was supported by the National Natural Science Foundation of China (No. 81671423 and No. 81402130), the National Key Research and Development Program of China (No. 2018YFC1003100), Liaoning Provincial Key Research and Development Program (No. 2018225090), the Fok Ying Tung Education Foundation (No. 151039) and Distinguished Talent Program of Shengjing Hospital (No. ME76). No competing interests declared.

scholarly journals Non-Invasive Prediction of Blastocyst Formation by Day Three Embryo Culture Medium Mass Spectrometry Lipid Fingerprinting

2015 ◽  
Vol 19 (3) ◽  
pp. 119-124
Author(s):  
Daniela Paes de Almeida Ferreira Braga ◽  
Amanda Souza Setti ◽  
Elaine Cristina Cabral ◽  
Marcos Eberlin ◽  
Edson Guimarães Loturco ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Embryo Culture ◽  
Culture Medium ◽  
Blastocyst Formation ◽  
Non Invasive ◽  
Medium Mass ◽  
Embryo Culture Medium

scholarly journals Evaluation of in vitro production rates of bovine embryos using melatonin-supplemented culture medium

2021 ◽  
Vol 10 (6) ◽  
pp. e19010615544
Author(s):  
Ricardo Magalhães ◽  
Carlos Renato de Freitas Guaitolini ◽  
Marcio Luiz Denck Tramontin ◽  
Danielle Andressa Oliveira Sestari ◽  
Bruno Argenton de Barros ◽  
...  
Keyword(s):  
Culture Medium ◽  
Statistical Significance ◽  
Gradient Centrifugation ◽  
Control Group ◽  
Bovine Embryo ◽  
Blastocyst Formation ◽  
In Vitro Production ◽  
Embryo Production ◽  
Vitro Fertilization

In this study, we aimed to evaluate the rate of bovine embryo production by using 50 ng/mL melatonin supplementation in in vitro culture medium. For this, oocytes from slaughterhouse ovaries were matured in vitro in TCM-199 medium with Earle’s balanced salt solution + 10% SFB, FSH, and LH in an atmosphere of 5% CO2. Twenty-four hours after IVM, the oocytes underwent in vitro fertilization in human tubal fluid under the same conditions as above, for 18 h. Semen was fractionated by Percoll gradient centrifugation and the concentration of sperm was adjusted to 1 × 106/mL. Probable zygotes were then divided into two groups: the control group grown in drops of 90 μL SOFaa medium + 0.6% BSA + 2.5% SFB, in an atmosphere of 5% CO2, 90% N2, and a melatonin group (Mel), similarly cultured in 90 μL drops of SOFaa medium + 0.6% BSA + 2.5% SFB + 50 ng/mL melatonin. Cleavage rates were assessed on day 3 (D3). On D7, blastocyst formation rates were evaluated. Eight routines were performed (320 oocytes per routine). Data were analyzed with ANOVA, followed by Tukey’s range test using a general linear model. The level of statistical significance was set at 5%. There were no differences in the rates of cleavage or blastocyst formation between the control and melatonin groups (P > 0.05). Thus, under the conditions used in this study, supplementation with melatonin did not yield benefits in increasing the rate of in vitro bovine embryo production.

291 EFFECTS OF ANTIOXIDANT AND CULTURE MEDIUM ON THE DEVELOPMENT OF PORCINE IN VITRO-MATURED - IN VITRO-FERTILIZED EMBRYOS

2007 ◽  
Vol 19 (1) ◽  
pp. 261
Author(s):  
C. Choe ◽  
D.-S. Son ◽  
S.-H. Choi ◽  
S.-R. Cho ◽  
H.-J. Kim ◽  
...  
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Culture Conditions ◽  
Free Oxygen Radicals ◽  
Blastocyst Formation ◽  
Free Oxygen ◽  
Statistical Analysis System ◽  
Developmental Rates ◽  
Analysis System

Most cells cultured in vitro are exposed to the risk of injury by free oxygen radicals (FOR). However, some of FOR-induced injury could be reduced by the antioxidants and culture medium used for in vitro embryos. This study was undertaken to examine the effects of the antioxidant and culture medium on the development of porcine in vitro-matured–in vitro-fertilized embryos. In Experiment 1, we treated the porcine oocytes in NCSU23 medium with various concentrations of β-mercaptoethanol (β-ME) to determine the effective concentration of antioxidants during IVM of porcine oocytes. In Experiment 2, we tested different culture media to find the proper culture conditions for in vitro porcine embryos. The porcine oocytes that were matured in NCSU23 medium and then fertilized in mTBM medium were cultured in NCSU23 or porcine zygote medium-5. All steps (maturation, fertilization, and development) were carried out in vitro. Differences were analyzed among treatments using the general linear model (GLM) procedure in the Statistical Analysis System (SAS Institute, Inc., Cary, NC, USA). The results were summarized as follows. Various concentrations of β-ME showed different developmental rates in porcine embryos. The rates of blastocyst formation at Day 7 after IVF were 9.2 � 1.8 (n = 65), 10.0 � 4.2 (n = 80), 17.5 � 1.1 (n = 63), 20.7 � 1.7 (n = 82), and 14.6 � 1.4 (n = 82) in oocytes treated with β-ME at 0, 10, 25, 50, and 100 �M during IVM, respectively. Of the concentrations of β-ME tested, 50 �M β-ME markedly increased the rates of blastocyst formation at Day 7 (P < 0.05). The rates of blastocyst formation at Day 7 in the NCSU23 and PZM-5 culture media of porcine IVF-derived embryos were 18.8 � 2.6 (n = 96) and 15.6 � 7.1 (n = 77), respectively. The developmental rates were slightly increased in NCSU23, compared with those in PZM-5, but there were no significant differences (P < 0.05) between the NCSU23 and PZM-5 media. In conclusion, these results suggest that the addition of 50 �M β-ME in the IVM medium can improve developmental the rates of porcine embryos in vitro.

Sign in / Sign up

Export Citation Format

Share Document