scholarly journals 279VASCULAR MORPHOMETRY OF BOVINE PLACENTOMES IN LATE GESTATION FROM EMBRYOS PRODUCED IN VIVO OR IN VITRO

2004 ◽  
Vol 16 (2) ◽  
pp. 259
Author(s):  
J.R. Miles ◽  
C.E. Farin ◽  
K.F. Rodriguez ◽  
J.E. Alexander ◽  
P.W. Farin
Keyword(s):  
Blood Vessels ◽  
Maternal Blood ◽  
Volume Density ◽  
Late Gestation ◽  
Embryo Production ◽  
Vascular Volume ◽  
Treatment Groups ◽  
In Vitro Embryo Production

The role of the vascular supply in the development of placentas from embryos produced in vitro is poorly understood. The objective of this study was to determine the effects of in vitro embryo production on morphometry of blood vessels within fetal (cotyledonary) and maternal (caruncular) components of the placentome during late gestation. In vivo-produced embryos were recovered from superovulated Holstein cows on Day 7 after estrus. For in vitro embryo production, oocytes were aspirated from the ovaries of Holstein cows, matured in vitro, and then fertilized. Presumptive zygotes with their cumulus cells were transferred into M-199 with 10% estrus cow serum and cultured for 168h post-insemination. Semen from the same Holstein sire was used for the production of in vivo and in vitro embryos. Single blastocysts from each production system were transferred into the uteri of heifers. On Day 222 of gestation, fetuses and placentas were recovered in utero (in vivo, n=12; in vitro, n=12). Placentomes were collected, fixed and sectioned. Fetal and maternal blood vessels were identified within placentome sections using immunocytochemistry for vascular endothelial growth factor (VEGF) protein. A total of 4.8×105μm2 of tissue were examined from each placentome. Stereological methods were used to determine the volume densities of fetal and maternal blood vessels. Data were analyzed by GLM procedures. Fetuses were heavier (P=0.03) in the in vitro group (20.7±1.0kg, LS mean±SEM) compared to the in vivo group (17.3±1.0kg). Placentas were also heavier (P=0.06) for the in vitro group (2.5±0.2kg) compared to the in vivo group (2.0±0.2kg). Placental efficiency, calculated as fetal weight/placental weight, was similar between the two treatment groups (9.0±0.5 and 8.9±0.5 for in vivo and in vitro, respectively). Fetal vascular volume density in placentomes was not different between the two treatment groups (5.4±0.3% and 5.4±0.3% for in vivo and in vitro, respectively). In contrast, maternal vascular volume density was greater (P=0.02) for placentomes in the in vitro group (5.9±0.3%) compared to in vivo controls (4.9±0.3%). In summary, compared to placentomes from embryos produced in vivo, placentomes from embryos produced in vitro had similar volume density of fetal vessels, but had significantly increased volume density of maternal vessels. Supported by the State of North Carolina.

scholarly journals Angiotensin II infusion in vivo does not modulate cortisol secretion in the late-gestation ovine fetus

1998 ◽  
Vol 275 (2) ◽  
pp. R357-R362 ◽  
Author(s):  
Kirsten R. Poore ◽  
I. Ross Young ◽  
Benedict J. Canny ◽  
Geoffrey D. Thorburn
Keyword(s):  
Angiotensin Ii ◽  
Adrenal Gland ◽  
Late Gestation ◽  
Treatment Groups ◽  
Arterial Blood ◽  
Basal Cortisol ◽  
Ovine Fetus

Maturation of the fetal adrenal gland is critical for the onset of ovine parturition. It has long been proposed that the fetal adrenal gland may be under inhibitory influences during late gestation. In vitro evidence has suggested that angiotensin II may be such an inhibitory factor and may help to prevent a premature increase in cortisol concentrations. The aim of this study was to test the effect of angiotensin II infusion in vivo on basal cortisol concentrations and fetal adrenal responsiveness to an ACTH-(1—24) challenge. Fetuses received a continuous infusion of either angiotensin II (100 ng ⋅ min−1 ⋅ kg−1; n = 7) or saline (2 ml/h; n = 4), which commenced at 140 days of gestation (GA) and continued for a total of 50 h. Adrenal responsiveness to the administration of ACTH-(1—24) (5 μg/kg) was determined during angiotensin II or saline infusions at both 2 and 48 h after infusion onset. Angiotensin II had no significant effect on adrenal responsiveness after acute (2 h) or chronic (48 h) infusion. There was no effect of saline or angiotensin II infusion on basal immunoreactive ACTH or cortisol concentrations after 2 h, but there was a significant increase in basal cortisol concentrations in both treatment groups by 48 h, probably reflecting the normal rise in cortisol concentrations at this GA. Mean arterial blood pressure was significantly increased in angiotensin II-infused fetuses only. This study has therefore found no evidence to suggest that angiotensin II infusion in vivo modulates fetal basal cortisol concentrations or adrenal responsiveness in the last week of gestation, in contrast with previous in vitro studies. These results throw into question the proposed role of angiotensin II as a negative modulator of adrenal function in the ovine fetus.

281 EVALUATION OF IN VITRO EMBRYO PRODUCTION EFFICIENCY USING SEXED BULL SPERM SORTED WITH TWO TYPES OF CELL SORTER

2011 ◽  
Vol 23 (1) ◽  
pp. 238
Author(s):  
H. Hayakawa ◽  
T.-I. Hirata
Keyword(s):  
Production Efficiency ◽  
Bos Taurus ◽  
Digital Processing ◽  
Embryo Production ◽  
Cell Sorter ◽  
Treatment Groups ◽  
Fort Collins ◽  
In Vitro Embryo Production ◽  
Bull Sperm

Cell sorting is an important part of the sperm sexing process. The objective of this study was to compare the efficiency of in vitro embryo production using sexed frozen–thawed bull sperm sorted with 2 types of cell sorter. Ejaculates from 2 Bos taurus (Holstein, 5 years old) bulls underwent conventional processing (control) or sorting for X chromosome bearing sperm using MoFlo® SX (SX, Dako, Fort Collins, CO, USA) or MoFlo® XDP-SX (XDP, Beckman Coulter, Fullerton, CA, USA) following XY™ sperm-sorting protocols. Processed sperm samples were cryopreserved in 0.5-mL plastic straws. Cumulus–oocyte complexes obtained from abattoir-derived ovaries were matured for 20 h in HEPES–TCM-199 (Lu and Seidel 2004 Theriogenology 62, 819–830) and randomly assigned to each of 3 sperm treatment groups. Thawed sperm were centrifuged for 20 min at 448 × g through an ISolate® (Irvine Scientific, Santa Ana, CA, USA) gradient (45:90%). Sperm pellets were washed in IVF100 (Hoshi 2003 Theriogenology 59, 675–685) by centrifugation for 5 min at 252 × g. Oocytes were co-incubated with washed sperm (5 to 10 × 106 sperm mL–1) in IVF100 (Hoshi 2003 Theriogenology 59, 675–685) for 8 h at 38.5°C in 5% CO2 and 95% air (Day 0). Presumptive zygotes were cultured for 90 h in CDM-1 (Lu and Seidel 2004 Theriogenology 62, 819–830) and then washed and cultured in IVD101 (Hoshi 2003 Theriogenology 59, 675–685) at 38.5°C in 5% CO2, 5% O2, and 90% N2. Cleavage rates on Day 2 and blastocyst rates on Day 7 to 9 were recorded after insemination. Two-way ANOVA was used for data analysis, followed by Fisher’s PLSD test. Experiments were replicated 4 times for bull A (total of 1 350 oocytes used) and 5 times for bull B (total of 1 529 oocytes used). The data are summarised in Table 1. No interaction was observed between the treatments and bulls. Cleavage rates were not significantly different in the 3 treatment groups. However, blastocyst rates were significantly lower in both SX (P < 0.001) and XDP (P < 0.002) groups than in control groups for both bulls but not different between SX and XDP (P > 0.8). Bull B showed significantly poorer results than bull A regarding both cleavage (P < 0.003) and blastocyst (P < 0.02) rates. MoFlo® SX (analogue processing) has been used for a decade, and XDP (digital processing) is the replacement model with its accelerated sorting speed. The current results indicated that the in vitro embryo production efficiency did not differ between sperm sorted with either SX or XDP. We suggest that sperm can be sorted using XDP without compromising sperm health. Table 1.Cleavage and blastocyst rates after IVF with 2 Holstein bulls for three sperm treatments

Predictive value of in vitro embryo production characteristics for in vivo fertility of bulls

1997 ◽  
Vol 47 (1) ◽  
pp. 259 ◽  
Author(s):  
L.M.T.E. Lansbergen ◽  
E.H.A.T. Hanenberg ◽  
A.M. van Wagtendonk-de Leeuw
Keyword(s):  
Predictive Value ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
Production Characteristics

97 IN VIVO AND IN VITRO EMBRYO PRODUCTION WITH Y-SEXED SORTED OR CONVENTIONAL SEMEN IN BEEF CATTLE

2014 ◽  
Vol 26 (1) ◽  
pp. 162
Author(s):  
H. Tribulo ◽  
J. Carcedo ◽  
R. Tribulo ◽  
J. Menajovsky ◽  
B. Bernal ◽  
...  
Keyword(s):  
Animal Health ◽  
T Test ◽  
High Fertility ◽  
In Vitro Production ◽  
Embryo Production ◽  
Sexed Semen ◽  
Chi Squared ◽  
In Vitro Embryo Production

An experiment was designed to evaluate in vivo and in vitro embryo production following the use of frozen–thawed conventional or Y-sexed semen from a Brangus bull with known high fertility. For in vivo embryo production, Brangus heifers (n = 12) were superovulated twice in a crossover design and inseminated with sexed or conventional semen. On Day 0, all heifers received an intravaginal progesterone device (DIB 1 g, Syntex S.A., Buenos Aires, Argentina) and 2.5 mg oestradiol benzoate and 50 mg progesterone (Progestar, Syntex S.A.) by intramuscular injection (IM). On Day 4, heifers were superstimulated with 200 mg of NIH-FSH-P1 Folltropin-V (Bioniche Animal Health, Belleville, Ontario, Canada) in twice-daily decreasing doses over 4 days. In the a.m. and p.m. of Day 6, all heifers received PGF2a (Ciclase, Syntex) and DIBs were removed in the p.m.. In the a.m. of Day 8, heifers received 100 μg de Gonadolerin (Gonasyn, Syntex S.A.) and were randomly allocated to receive either one straw of conventional semen (24 × 106 sperm per dose) 12 and 24 h later or two straws of sexed semen (2.4 × 106 sperm per dose) 18 and 24 h after GnRH. Ova/embryos were collected nonsurgically on Day 15 and evaluated following IETS recommendations. Means were compared by t-test. Mean ( ± s.e.m.) number of ova/embryos, fertilized ova, and transferable embryos were 14.8 ± 2.7, 9.4 ± 1.8, and 7.1 ± 1.7 v. 16.8 ± 3.1, 9.9 ± 2.5, and 8.1 ± 2.0 for donors inseminated with conventional or sexed semen, respectively (P > 0.6). For in vitro production, oocytes were obtained from 50 ultrasound-guided follicle aspiration (OPU) sessions that was performed at random stages of the oestrous cycle and without superstimulation in 22 Brangus cows and heifers. Oocytes were classified and matured in TCM-199 medium with NaHCO3 and supplemented with 1% fetal bovine serum. Semen samples from the same bull used for in vivo embryo production were selected using Percoll and capacitated in Fert medium and used at a final concentration of sperm/mL for nonsexed semen and 2 × 106 sperm mL–1 for sexed semen. After 16 h (sexed) or 18 h (conventional) in Fert medium, zygotes were denuded and cultured in SOF supplemented with 0.4% BSA under oil at 37°C, 5% CO2 and saturated humidity for 7 days. The total number of oocytes matured and fertilized was 528 and 318 for conventional and sexed semen, respectively. Means were compared by t-test and proportions by chi-squared test. Mean (± s.e.m.) number of cleaved zygotes and blastocysts produced per OPU session did not differ between conventional (11.0 ± 1.4 and 7.1 ± 1.0) and sexed (8.7 ± 0.8 and 4.9 ± 0.7; P > 0.2) semen. However, the proportion of cleaved zygotes and blastocysts produced were significantly higher (P < 0.05) with conventional semen (61.2%; 329/538 and 39.4%; 212/538) than with sexed semen (54.4%; 173/318 and 30.8%; 98/318), respectively. In conclusion, comparable number of embryos can be obtained in vivo with sexed or conventional semen from a bull with proven high fertility. However, the proportion of blastocysts produced in vitro is likely to be reduced following the use of sexed as compared with conventional semen from the same bull.

135 Use and dose of porcine follicle-stimulating hormone for ovarian superstimulation prior to ovum pickup and in vitro embryo production in pregnant Holstein heifers

2019 ◽  
Vol 31 (1) ◽  
pp. 192
Author(s):  
R. V. Sala ◽  
L. C. Carrenho-Sala ◽  
M. Fosado ◽  
E. Peralta ◽  
D. C. Pereira ◽  
...  
Keyword(s):  
Limited Information ◽  
Dominant Follicle ◽  
Blastocyst Development ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Oocyte Competence ◽  
Treatment Groups ◽  
Oocyte Recovery ◽  
In Vitro Embryo Production

The benefit of superstimulation with exogenous FSH before ovum pickup for in vitro embryo production has been the subject of significant controversy. In addition, there is limited information on different dose regimens. Thus, the objective of the present study was to evaluate the effect of dose of porcine (p)-FSH during superstimulation before ovum pickup (OPU) on in vitro embryo production in pregnant heifers. Pregnant Holstein heifers (n=36) were assigned to a complete 3×3 crossover design. Three treatment groups were evaluated as follows: p-FSH 0mg (FSH0), p-FSH 160mg (FSH160) and p-FSH 300mg (FSH300). Three sessions of OPU were performed on each animal at 48, 62 and 76 days of gestation, with a washout interval between sessions of 14 days. Follicular wave emergence was synchronized by dominant follicle removal. Heifers in the FSH0 group received no further treatment, whereas the remaining groups received a total of 4 injections 12h apart as follows: FSH160 (48.0, 42.7, 37.3 and 32.0mg) or FSH300 (90.0, 80.0, 70.0 and 60.0mg), beginning 36h after dominant follicle removal. Ovum pickup was performed in all heifers 40h after the last p-FSH injection. Heifers were subjected to OPU for oocyte recovery, and number of follicles was determined. Recovered oocytes were processed and in vitro embryo production performed. Differences between treatment groups were evaluated by generalized linear mixed models. Data are presented (Table 1) as mean±standard error of the mean. There was no effect of days in gestation for any of the outcomes evaluated (P&gt;0.05). Follicle numbers at the time of oocyte recovery were different (P&lt;0.01) between groups. Heifers in the FSH300 group had a greater (P&lt;0.05) number of medium, large and total follicles than heifers in the FSH0 group, whereas heifers in the FSH160 were intermediate. Total number of recovered, viable and cleaved oocytes were greater (P&lt;0.01) in FSH300- than in FSH160- and FSH0-treated heifers. Cleavage rate and blastocyst development rate were not different (P&gt;0.10) between groups. The number of grade 1 and 2 blastocysts was greater in FSH300- than in FSH160- and FSH0-treated heifers (P&lt;0.03). In summary, the use of 300mg of p-FSH before OPU in pregnant heifers increases the number of follicles, oocytes and blastocysts produced per heifer with no detrimental effect on oocyte competence. Table 1.Ovum pickup and in vitro embryo production in pregnant heifers treated with different doses of porcine FSH

Pregnancy obtained in a late gestational mare by in vitro embryo production

2019 ◽  
Vol 31 (12) ◽  
pp. 1926
Author(s):  
Lino Fernando Campos-Chillon ◽  
Jan Martin ◽  
Joy L. Altermatt
Keyword(s):  
Transvaginal Ultrasound ◽  
Experimental Studies ◽  
Late Gestation ◽  
Follicular Growth ◽  
Ultrasound Guided ◽  
Embryo Production ◽  
In Vitro Embryo Production

Recently, the demand for invitro embryo production in the horse has increased worldwide. Most clinical transvaginal ultrasound-guided ovum pick-up (OPU) procedures are performed in non-pregnant donor mares, and few experimental studies have described invitro embryo production from oocytes of pregnant donors 21–150 days in gestation. This report discusses OPU, follicular growth and invitro embryo production in a pregnant mare during late gestation.

In vivo oocyte recovery and in vitro embryo production from bovine donors aspirated at different frequencies or following FSH treatment

1999 ◽  
Vol 51 (5) ◽  
pp. 951-961 ◽  
Author(s):  
K.L Goodhand ◽  
R.G Watt ◽  
M.E Staines ◽  
J.S.M Hutchinson ◽  
P.J Broadbent
Keyword(s):  
Embryo Production ◽  
Oocyte Recovery ◽  
In Vitro Embryo Production

In vitro embryo production compared to in vivo production using gamete recovery and follicular transfer (GRAFT)

1997 ◽  
Vol 47 (1) ◽  
pp. 287
Author(s):  
P. Davies ◽  
J. Singh ◽  
J. Thundathil ◽  
G. Brogliatti ◽  
D. Bergfelt ◽  
...  
Keyword(s):  
Embryo Production ◽  
In Vitro Embryo Production
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