scholarly journals 233.Ammonium affects mitochondrial distribution and function in mouse 2-cell embryos

2004 ◽  
Vol 16 (9) ◽  
pp. 233 ◽  
Author(s):  
D. L. Zander ◽  
D. A. Froiland ◽  
M. Lane
Keyword(s):  
Amino Acids ◽  
Membrane Potential ◽  
Intensity Ratio ◽  
Culture Media ◽  
Blastocyst Development ◽  
Cell Stage ◽  
Pixel Intensity ◽  
Essential Components ◽  
Mitochondrial Distribution

Amino acids are key regulators of embryo function and are essential components in embryo culture media. Amino acids spontaneously breakdown and are metabolised by embryos resulting in ammonium build-up in the medium. While ammonium does not affect blastocyst development, the ability of these blastocysts to implant was reduced along with subsequent fetal growth rates. However, the mechanism for the inhibitory effect of ammonium is currently not known. It has been demonstrated in other tissues that mitochondrial bioenergetics can be disrupted by the presence of ammonium in the media which subsequently affects cellular viability. Therefore, the aim of this study was to examine the effects of ammonium on the mitochondria of mouse embryos cultured in the presence of ammonium. Mouse zygotes from superovulated females were cultured in medium G1.2 with or without 300 μM ammonium for 22 h at 37oC in 6%CO2�:�5%O2�:�89%N2. In vivo-developed 2-cell embryos were flushed from the reproductive tract and assessed immediately. At the 2-cell stage mitochondrial distribution (Mitotracker) and membrane potential (JC-1) were assessed using confocal microscopy and images were quantitated using IP Lab software package. Differences between treatments were determined using ANOVA and Bonferroni's multiple comparison procedure. Culture of zygotes to the 2-cell stage in medium G1.2 did not affect mitochondrial distribution compared to in vivo controls. However, 2-cell embryos cultured with ammonium had a decrease in their mitochondrial nuclear�:�cortical ratio (97���1 compared to 106���1; P�<�0.05) indicating that mitochondria were dispersing away from the nuclei. Culture with ammonium also significantly decreased the mitochondrial membrane potential (0.50���0.01 mean pixel intensity ratio) compared to those cultured without ammonium (0.72���0.3 mean pixel intensity ratio, P�<�0.001). The data presented demonstrates that culture for only 24�h with ammonium disrupts both mitochondrial distribution and membrane potential and supports our hypothesis that mitochondria are an early target for the inhibitory action of ammonium.

257. Addition of glycine to vitrification solutions protects oocyte and embryo physiology and health

2005 ◽  
Vol 17 (9) ◽  
pp. 104
Author(s):  
K. S. Cashman ◽  
D. A. Froiland ◽  
J. G. Thompson ◽  
M. Lane
Keyword(s):  
Gene Expression ◽  
Membrane Potential ◽  
Mitochondrial Membrane ◽  
Multiple Comparisons ◽  
Developmental Competence ◽  
Rt Pcr ◽  
Blastocyst Development ◽  
Pixel Intensity ◽  
Mitochondrial Distribution

Cryopreservation procedures for oocytes result in a significant reduction in viability. Although cryopreservation procedures cause dehydration and therefore osmotic stress, the role of osmolytes in solutions has not been considered and they have therefore not been included for routine use. The aim of this study was to assess the effects of the addition of the osmolyte glycine to vitrification solutions on the health and developmental competence of mouse oocytes. Oocytes were collected from F1 female mice and cryopreserved using cryoloop vitrification with or without glycine, with fresh oocytes examined as controls (n = 2086). Mitochondrial distribution and membrane potential as well as the morphology of the spindles and chromosomes were assessed. Oocytes were fertilised to assess their ability to develop into blastocysts, which were then assessed for their expression of Glut1, Glut3 and IGF2 by real-time RT-PCR. Statistical analysis was performed using a generalised linear model followed by multiple comparisons using an LSD test. Vitrification without glycine perturbed mitochondrial distribution (mean pixel intensity of outer region:inner region, 1.58±0.20, P<0.01) and mitochondrial membrane potential (mean pixel intensity 0.56±0.01, P<0.01) compared to control oocytes (2.34±0.24 and 0.52±0.01, respectively). The addition of glycine prevented these changes (1.97±0.16 and 0.53±0.01, respectively). Vitrification without glycine resulted in 52% of spindles and chromosomes appearing normal while this was increased to 69% with the addition of glycine, however in both treatments these abnormalities appeared to recover after culture for 2 h. Vitrification did not affect fertilisation and blastocyst development however expression of Glut3 was decreased 2.9 fold in blastocysts resulting from oocytes vitrified in the absence of glycine (P<0.01). The data presented suggests that the addition of glycine results in fewer perturbations in oocyte physiology and gene expression of the subsequent blastocysts and should therefore be considered for routine inclusion in solutions for the cryopreservation of oocytes.

Insertions of up to 17 amino acids into a region of alpha-tubulin do not disrupt function in vivo

1987 ◽  
Vol 7 (10) ◽  
pp. 3799-3805
Author(s):  
P J Schatz ◽  
G E Georges ◽  
F Solomon ◽  
D Botstein
Keyword(s):  
Amino Acids ◽  
Variable Region ◽  
Yeast Cells ◽  
Meiotic Spindle ◽  
Nuclear Movement ◽  
Yeast Saccharomyces Cerevisiae ◽  
Alpha Tubulin ◽  
Essential Components ◽  
Altered Proteins

Microtubules in yeasts are essential components of the mitotic and meiotic spindle and are necessary for nuclear movement during cell division and mating. The yeast Saccharomyces cerevisiae has two alpha-tubulin genes, TUB1 and TUB3, either of which alone is sufficient for these processes when present in a high enough copy number. Comparisons of sequences from several species reveals the presence of a variable region near the amino terminus of alpha-tubulin proteins. We perturbed the structure of this region in TUB3 by inserting into it 3, 9, or 17 amino acids and tested the ability of these altered proteins to function as the only alpha-tubulin protein in yeast cells. We found that each of these altered proteins was sufficient on its own for mitotic growth, mating, and methods of yeast. We conclude that this region can tolerate considerable variation without losing any of the highly conserved functions of alpha-tubulin. Our results suggest that variability in this region occurs because it can be tolerated, not because it specifies an important function for the protein.

scholarly journals Mitochondrial membrane potential in 2-cell stage embryos correlates with the success of preimplantation development

Reproduction ◽  
2014 ◽  
Vol 147 (5) ◽  
pp. 627-638 ◽  
Author(s):  
Kouji Komatsu ◽  
Akira Iwase ◽  
Miki Mawatari ◽  
Jingwen Wang ◽  
Mamoru Yamashita ◽  
...  
Keyword(s):  
Membrane Potential ◽  
Mitochondrial Membrane Potential ◽  
Mitochondrial Membrane ◽  
Developmental Process ◽  
Preimplantation Embryos ◽  
Cell Stage ◽  
Hormonal Stimulation ◽  
Maternal Mrna

Hormonal stimulation in superovulation induces female mice to ovulate more oocytes than spontaneous ovulation. Because the superovulated oocytes contain a number of oocytes that normally regress before spontaneous ovulation or immature oocytes, the development of some embryos that derive from these oocytes by IVF is prevented. Therefore, the quality of superovulated oocytes should differ from that of spontaneously ovulated oocytes. In this study, we evaluated the quality of superovulated oocytes, by examining 1- and 2-cell stage embryos, in which the development mainly depends on the maternal mRNA, proteins, and mitochondria that are contained in the oocytes, and we then measured the mitochondrial membrane potential (ΔΨm) of the 1- and 2-cell stage,in vivo-fertilized, and IVF embryos. The ΔΨmof 1-cell stage IVF embryos was lower than that ofin vivo-fertilized embryos; however, there was no difference between IVF embryos. During the developmental process from 1- to 2-cell stage, the ΔΨmofin vivo-fertilized embryos was highly upregulated, whereas a number of IVF embryos remained unchanged. As a result, 2-cell stage embryos were divided into two groups: high- and low- ΔΨm2-cell stage IVF embryos. The development of low-ΔΨm2-cell stage IVF embryos tended to be arrested after the 2-cell stage. These results indicated that the upregulation of ΔΨmduring the 1- to 2-cell stage was important in the development of early preimplantation embryos; there were some defects in the mitochondria of superovulated oocytes, which prevented their development.

Expression of mRNA encoding leukaemia inhibitory factor (LIF) and its receptor (LIFRβ) in buffalo preimplantation embryos produced in vitro: markers of successful embryo implantation

Zygote ◽  
2012 ◽  
Vol 21 (2) ◽  
pp. 203-213 ◽  
Author(s):  
S. Eswari ◽  
G. Sai Kumar ◽  
G. Taru Sharma
Keyword(s):  
Culture Media ◽  
Total Cell ◽  
Inhibitory Factor ◽  
Preimplantation Embryos ◽  
Leukaemia Inhibitory Factor ◽  
Hatching Rate ◽  
Blastocyst Development ◽  
Cell Stage ◽  
Significant Difference

SummaryThe objective of this study was to evaluate the effect of supplementation of recombinant leukaemia inhibitory factor (LIF) in culture media on blastocyst development, total cell number and blastocyst hatching rates and the reverse transcription-polymerase chain reaction analysis of preimplantation buffalo embryos to determine whether they contain the LIF-encoding mRNA and its beta receptor (LIFRβ) genes in different stages of preimplantation buffalo embryos. Cumulus–oocyte complexes retrieved from slaughterhouse buffalo ovaries were matured in vitro and fertilized using frozen buffalo semen. After 18 h of co-incubation with sperm, the presumptive zygotes were cultured in modified synthetic oviductal fluid without (control) or with rhLIF (100 ng/ml). There was no significant difference in the overall cleavage rate up to morula stage however the development of blastocysts, hatching rate and total cell numbers were significantly higher in the LIF-treated group than control. Transcripts for LIFRβ were detected from immature, in vitro-matured oocytes and in the embryos up to blastocyst stage, while transcripts for the LIF were detected from 8–16-cell stage up to blastocyst, which indicated that embryo-derived LIF can act in an autocrine manner on differentiation process and blastocyst formation. This study indicated that the addition of LIF to the embryo culture medium improved development of blastocysts, functional (hatching) and morphological (number of cells) quality of the blastocysts produced in vitro. The stage-specific expression pattern of LIF and LIFRβ mRNA transcripts in buffalo embryos indicated that LIF might play an important role in the preimplantation development and subsequent implantation of buffalo embryos.

scholarly journals Amino Acid Metabolism in Human Embryos

2016 ◽  
pp. 823-832 ◽  
Author(s):  
P. DRÁBKOVÁ ◽  
L. ANDRLOVÁ ◽  
R. HAMPL ◽  
R. KANĎÁR
Keyword(s):  
Amino Acids ◽  
Amino Acid ◽  
Amino Acid Metabolism ◽  
Culture Medium ◽  
Acid Metabolism ◽  
Culture Media ◽  
Time Lapse ◽  
Human Embryos ◽  
Strongly Correlated ◽  
Cell Stage

The aim of this study was to find some relationship between amino acid metabolism and the embryo morphokinetic parameters studied via time-lapse analysis. Study included 48 human embryo samples and their culture media. Two groups of embryos were identified: embryos reached the 8-cell stage on day 3 (n=34) and embryos failed to develop at any point during the incubation (n=14). Amino acids levels were measured on day 3 of embryo development; using time-lapse analysis, the precise timing of embryo cleavage, synchrony of division, grade of fragmentation etc. were established. No statistically significant differences between dividing and arresting embryos were observed in terms of amino acids production/consumption and turnover. Amino acids which were part of the culture medium did not exhibit any statistically significant correlation with kinetic parameters with the exception of the grade of fragmentation on day 3; there were negative correlation with glutamate, and positive with glutamine, glycine and taurine. In some dividing and in some arresting embryos appeared new amino acids which strongly correlated with each other, with methionine, but not with any other amino acid that is a regular part of the culture medium.

179 EFFECT OF SERUM DURING CULTURE ON DAY 14 ELONGATED BOVINE EMBRYOS

2011 ◽  
Vol 23 (1) ◽  
pp. 191 ◽  
Author(s):  
J. Angulo ◽  
G. T. Gentry ◽  
R. A. Godke ◽  
K. R. Bondioli
Keyword(s):  
Gene Expression ◽  
Messenger Rna ◽  
Culture Media ◽  
Calf Serum ◽  
Blastocyst Development ◽  
Cleavage Rate ◽  
Expression Levels ◽  
The Mean

It has been reported that the addition of serum to embryo culture media alters gene expression and triggers the development of large offspring syndrome. The objectives of this study were to determine gene expression levels in embryos cultured in the absence or presence of 5% calf serum and in vivo-derived (IVD) embryos and to determine the effects of serum on the length of elongated embryos. Abattoir-derived oocytes were obtained from a commercial provider and fertilized at 24 h of maturation with semen from a bull previously used for IVF. At 18 h post-insemination (hpi), embryos were denuded and groups of 15 presumptive zygotes were cultured in 30-μL drops of modified SOF medium with amino acids and 6 mg mL–1 of BSA (mSOFaa). At 72 hpi, cleavage rate was assessed and embryos were randomly allocated into 2 treatments: mSOFaa without and with 5% calf serum. Embryos were then cultured to 168 hpi and blastocyst rates were assessed and recorded. Blastocysts (n = 5 to 10) from each treatment were transferred into synchronized recipients, and Day 14 embryos were recovered 7 days post-transfer. Embryos were photographed, measured, and immediately stored at –80°C in a minimal volume of PBS + 0.1% polyvinyl alcohol. Messenger RNA was isolated using a Dynabeads mRNA Direct Kit™ (Invitrogen, Carlsbad, CA), and reverse transcription was performed using an iScript™ cDNA Synthesis Kit (Bio-Rad Laboratories, Inc., CA). Quantitative PCR was performed to determine the transcript abundance for COX6A, IFNT1a, PLAC8, IGF2R, and GAPDH for each sample. The GAPDH was used as a reference gene, and gene expression was calculated as a ratio of expression levels between each gene of interest and GAPDH. Expression levels for each gene were determined from standard curves generated by serial dilutions of PCR amplicons starting with 0.4 pg/reaction. Blastocyst development rates were higher in embryos cultured with serum compared with the nonserum treatment (14.9 and 7.4% respectively; chi-square, P < 0.001). Lengths of elongated embryos from the serum (3395.3 ± 414.7 μm) and nonserum (2784 ± 741.8 μm) culture treatments differed from the IVD (6297.7 ± 677.2 μm) treatment (mean ± SE; ANOVA, P < 0.0052). There were no differences in the mean expression levels for COX6A, IFNT1a, PLAC8, and IGF2R across treatment groups, but in the serum treatment, 3 out 11 overexpressed IFNT1a, 4 out of 11 overexpressed IGF2R, and 2 out of 11 overexpressed PLAC8, defined as being 2 standard deviations above the mean of the IVD treatment for each respective gene. In the in vitro-produced nonserum and IVD treatments, overexpression by this definition was not observed. Although mean expression levels were not affected by culture with serum under these conditions, very high expression of IFNT1a, IGF2R, and PLAC8 was observed in some embryos cultured with serum, but not in embryos cultured without serum or IVD embryos.

261. Disruption of mitochondrial function in the blastocyst alters expression of the chromatin remodeler ATRX

2008 ◽  
Vol 20 (9) ◽  
pp. 61
Author(s):  
S. L. Wakefield ◽  
A. N. Filby ◽  
M. Lane ◽  
M. Mitchell
Keyword(s):  
Mitochondrial Function ◽  
Metabolic Regulation ◽  
Culture Media ◽  
Maternal Diet ◽  
Cell Number ◽  
Early Embryo ◽  
Placental Development ◽  
Blastocyst Development ◽  
Cell Stage ◽  
Embryo Metabolism

Exposure of an embryo to suboptimal environments, including poor embryo culture media or inadequate maternal diet, can disrupt fetal and placental development and whilst the exact mechanisms responsible remain unknown, perturbed embryo metabolism has been implicated. We propose that stress applied to an early embryo causes mitochondrial dysfunction, resulting in a permanent epigenetic change. Thus the aim of this study was to determine the affect of directly perturbing mitochondria in the embryo, on development, metabolism and expression of the ATP-dependant chromatin remodelling protein, ATRX. Zygotes collected from gonadotrophin stimulated C57BL/6xCBA mice were cultured to the two-cell stage and then exposed to one of three treatments; control medium (C), medium lacking pyruvate (-P; embryos dependant on the mitochondrial Malate Aspartate Shuttle, MAS) or medium lacking pyruvate plus 5µM amino-oxyacetate (AOA), a specific MAS inhibitor (-P+AOA). Blastocyst development and metabolism were assessed by determining cell number and allocation, glycolysis, and ATP:ADP ratio. Relative gene expression of ATRX, was examined using RT PCR. Embryos dependant on the MAS alone (-P) had significantly decreased blastocyst development (87.1% v. 98.2%, P < 0.05), with a compensatory increase in glycolysis (0.20 v. 0.07 pmol/cell/hr, P < 0.001) despite a decrease in ATP:ADP (0.10 v. 0.13, P < 0.06), relative to the control. Inhibition of the MAS (-P+AOA) further reduced blastocyst development,(77.3%, P < 0.001) and decreased ATP:ADP (0.08, P < 0.004), but there was no change in glycolysis relative to control embryos (0.09 pmol/cell/hr, P = 0.3). Expression of ATRX was significantly increased for –P+AOA embryos relative to the control (1.63 v. 1.0, P < 0.007) but did not differ for –P embryos (1.1). This study demonstrates that direct perturbations of mitochondrial function in the embryo compromises its metabolic regulation and blastocyst development, and the expression of the epigenetic modulator ATRX. Further studies are underway to elucidate the implications of disrupted metabolic control and this epigenetic modulator on pregnancy outcomes.

Alteration of extracellular cation concentrations and ratios in culture medium does not affect first cleavage division of hamster zygotes in vitro nor overcome the 'two-cell block'

1989 ◽  
Vol 1 (3) ◽  
pp. 231 ◽  
Author(s):  
BD Bavister ◽  
M Golden
Keyword(s):  
Culture Medium ◽  
Culture Media ◽  
Cell Block ◽  
Cleavage Division ◽  
Cell Stage ◽  
Wide Range ◽  
Standard Culture ◽  
Simple Medium

In vivo fertilized hamster one-cell eggs (embryos) were cultured in a simple medium that was modified to provide a wide range of concentrations and ratios of the four major cation components (sodium, potassium, calcium and magnesium) while maintaining total osmotic pressure at 290 +/- 5 mosm. Embryos were cultured in these media to find the optimum cation concentrations for supporting the first cleavage division in vitro and to determine if physiologically abnormal cation concentrations and/or ratios in standard culture media could account for the 'two-cell block' to development in vitro in this species. Despite using a broad range of ratios for sodium:potassium (from 45:1 to 5:1) and for calcium:magnesium (from 17:1 to 1:1), there were no significant differences in the proportions of fertilized eggs that underwent the first cleavage division (approx. 60-80% across all treatments), and none of the two-cell embryos underwent further cleavage during extended culture. These data demonstrate that the first cleavage division of hamster embryos in vitro is insensitive to extracellular concentrations and ratios of the major cations, and that the non-physiological concentrations and/or ratios of these cations in the culture medium are not the primary reason for the failure of hamster zygotes to develop past the two-cell stage in vitro.

103 BIRTH OF CLONED PIGLETS DERIVED FROM AN OPTIMIZED IN VITRO BLASTOCYST PRODUCTION SYSTEM BY TRICHOSTATIN A TREATMENT

2007 ◽  
Vol 19 (1) ◽  
pp. 168
Author(s):  
Y.-H. Zhang ◽  
Y.-T. Du ◽  
K. Zhang ◽  
J. Li ◽  
P. M. Kragh ◽  
...  
Keyword(s):  
Culture Media ◽  
Trichostatin A ◽  
Blastocyst Stage ◽  
Control Group ◽  
Blastocyst Development ◽  
Cell Stage ◽  
Deacetylase Inhibitor ◽  
The One ◽  
Effective Group

The present study was designed to examine the effect of trichostatin A (TSA, a histone deacetylase inhibitor) treatment on in vitro developmental ability of pig cloned embryos and to evaluate the feasibility of producing piglets from these embryos. Cell lines were established from 40-day-old fetuses, and adult ear skin was used as nuclear donor. In vitro-matured oocytes from abattoir-derived sow ovaries were used as cytoplast recipients for micromanipulator-assisted somatic cell nuclear transfer (SCNT). Data were analyzed by using SPSS (11.0) with one-way ANOVA, and each experiment was replicated at least 3 times. In Experiment 1, immediately after simultaneous fusion and activation, the reconstructed couplets were randomly cultured in porcine zygote medium 3 (PZM3; Yoshioka et al. 2002 Biol. Reprod. 66, 112–119) with 10 �g mL-1 cytochalasin B (CB), 10 �g mL-1 cycloheximide (CHX), and 0 nM, 5 nM, or 50 nM TSA for the first 4 h. Cloned embryos (fused reconstructed couplets) were moved to the same culture media but without CB and CHX and further cultured at 38.5�C, under 5% CO2, 5% O2, 90% N2 and 100% humidity. After incubation for a total of 8–14 h in 50 nM, 19–24 h in 50 nM or 5 nM, and 31–36 h in 50 nM TSA in PZM3 (0 nM TSA serves as control for each group), the embryos were further cultured in vitro without TSA in PZM3 for up to 168 h. Cleavage and blastocyst development rates, based on embryos cultured, were recorded at 48 and 168 h of IVC, respectively. Results showed that 50 nM TSA treatment for 19-24 h supported a higher blastocyst development rate than the control group [No. blastocysts/No. embryos cultured (mean � SEM): 107/258, 47.4 � 5.9% vs. 65/324, 20.0 � 2.3%, respectively; P &lt; 0.05], whereas similar pre-implantation development was obtained between the other 3 test groups and the control. In Experiment 2, TSA-treated cloned embryos at the one-cell stage or blastocyst stage were transferred to recipients to examine the possibility of producing piglets. Ten cloned piglets (2 are healthy and 8 died shortly after birth) and one ongoing pregnancy were obtained from 3 recipients who received an average of 110 one-cell stage embryos, whereas 4 piglets originating from traditional cloning were produced from one recipient which received 28 traditional cloned blastocysts (produced from the effective group in Experiment 1) and 30 handmade but non-TSA-treated ones. Our data demonstrate that TSA treatment after SCNT in porcine can significantly improve the in vitro blastocyst production, and embryos treated with TSA could support full-term development and result in healthy offspring.

86 IN VITRO-MATURED GILT OOCYTES CAN HAVE EQUAL OR BETTER DEVELOPMENTAL COMPETENCE THAN SOW OOCYTES WITH NEW MATURATION MEDIA

2017 ◽  
Vol 29 (1) ◽  
pp. 150 ◽  
Author(s):  
L. D. Spate ◽  
S. L. Murphy ◽  
J. A. Benne ◽  
A. Giraldo ◽  
D. Hylan ◽  
...  
Keyword(s):  
Growth Factor ◽  
Somatic Cell ◽  
Culture Media ◽  
Blastocyst Stage ◽  
Developmental Competence ◽  
Blastocyst Development ◽  
Development In Vitro ◽  
Humidified Air

It has long been thought that oocytes obtained from sows yielded a higher level of developmental competence compared with oocytes obtained from prepubertal gilts. Because gilt-derived oocytes are more readily available to our laboratory and they are less developmentally competent, we hypothesised that by making alterations to our maturation system we could improve the developmental competence of the gilt-derived oocytes to that of their sow-derived counterparts. We performed 2 experiments that evaluated the ability of each source of oocyte to develop to the blastocyst stage, using altered maturation media. The first experiment focused on the developmental ability of each source of oocytes, through IVF and culture. The second experiment again focused on the developmental competence of each oocyte source but through somatic cell NT. For both experiments, the sow-derived oocytes were obtained from Desoto Biosciences and the gilt ovaries were collected from Smithfield Inc. in Milan, Missouri. Both sets of oocytes were in vitro matured in M199 supplemented with 0.57 mM cysteine, 5 μg mL−1 LH and FSH, and 10 ng mL−1 epidermal growth factor; however, the gilt derived media was altered to contain 40 ng mL−1 fibroblast growth factor 2 and 20 ng mL−1 insulin-like growth factor and leukemia inhibitory factor. Additionally, the maturation media for the sow-derived oocytes contained the addition of 5 μg mL−1 insulin and 10% follicular fluid. In the first experiment we performed IVF on oocytes from the 2 sources as per our laboratory standard IVF procedure, co-incubating the oocytes with 0.25 × 106 porcine semen for 4 h, followed by washing and moving the oocytes to MU2 culture media at 38.50°C in 5% CO2, humidified air overnight. After overnight culture the presumptive zygotes were transferred to the same conditions with 5% CO2, 5% O2, and 90% N2. After an additional 5 days, blastocyst development was assessed. The gilt oocytes yielded 39.3a ± 7.2% blastocyst, and the sow oocytes had a blastocyst rate of 24.9b ± 6.9%, with an n of 389 and 313, respectfully. Statistical analysis was performed by using Genmod in SAS 9.4. In the second experiment, using standard laboratory protocol for somatic cell NT, we activated both sets of oocytes with 200 μM thimerosal for 10 min followed by 30-min incubation with 4 mM dithiothreitol. The embryos were co-incubated for 15 h with 500 nM Scriptaid in the MU2 culture media in 5% CO2, humidified air; then these embryos were also moved to 5% CO2, 5% O2, and 90% N2 and cultured to Day 6. The sow oocytes produced a blastocyst percentage of 38.6%, and the gilt oocyte group had a blastocyst percentage of 43.5%, with an n of 290 and 285, respectfully. There was no difference statistically between these treatments. Both gilt and the sow oocyte sources have yielded live piglets at this time. We concluded that the maturation system used for our gilt-derived oocytes resulted in equal or better development in vitro compared with the sow-derived oocytes. Follow-up experiments evaluating in vivo development are needed for a complete comparison. This work was funded by Food for the 21st Century University of MO, and the NIH U42OD011140.

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