scholarly journals Effect of Adding Insulin Transferrin Selenium (ITS) in the Medium on Maturation and Fertilization Rates of Bali Cattle Oocytes

2019 ◽  
Vol 24 (3) ◽  
pp. 95
Author(s):  
Hikmayani Iskandar ◽  
H Sonjaya ◽  
M Yusuf
Keyword(s):  
Germinal Vesicle ◽  
Fertilization Rate ◽  
Fertilization Rates ◽  
Randomized Design ◽  
Maturation Rate ◽  
Bali Cattle ◽  
Germinal Vesicle Break Down ◽  
Metaphase I

The aim of this study was to determine the effect of adding Insulin Transferrin Selenium (ITS) in the medium on maturation and fertilization rates of Bali cattle oocytes. The ovarium of Bali cows were sliced to produce oocytes, then the collected oocytes were subsequently selected based on their quality. Those oocytes were maturated for 24 hours and then fertilized for 18 hours in the incubators of 5% CO2 with temperature of 38.5ºC. Oocytes were stained using 2% of aceto orcein, and observed under a microscope. This study was divided into four treatments at different addition of ITS in the medium and replicated four times, and analized using a Factorial Complete Randomized Design (CRD)at each treatment. The treatments consisted of P0 as control, P1 (5 ng/ml), P2 (10 ng/ml), and P3 (15 ng/ml). Parameters observed in this study were the stages of oocytes maturation rate that consisted of germinal vesicle (GV), germinal vesicle break down (GVBD), metaphase I (M-I), and metaphase II (M-II), and fertilization rate that consisted of absence of pronucleus (P0), 1 pronucleus (PN1), 2 pronucleus (PN2), and more than two pronucleus (>2PN). The results of this study showed that addition of ITS at 5-15 ng/ml did not effect maturation and fertilization rates of Bali cows oocytes.The conculision of the study is addition of ITS at 5 ng / ml produces the best level of maturation and for the best level of fertilization with addition ITS at 15 ng / ml.

Fertilization and early embryonic development of in vitro matured metaphase I oocytes in patients with unexpected low oocyte maturity rate

Zygote ◽  
2021 ◽  
pp. 1-5
Author(s):  
Nafiye Yılmaz ◽  
Şebnem Özyer ◽  
Derya Taş ◽  
Mehmet Caner Özer ◽  
Ayten Türkkanı ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Fertilization Rate ◽  
Early Embryonic Development ◽  
Oocyte Maturity ◽  
Significant Difference ◽  
Maturation Rate ◽  
Developmental Capacity ◽  
Metaphase I

Summary To determine the fertilization and embryonic potential of immature metaphase I (MI) oocytes in patients with low oocyte maturity rate in whom the percentage of mature oocytes obtained was less than 75% of the total retrieved ones. In vivo matured metaphase II (MII) oocytes (MII-ICSI, n = 244), and in vitro matured MI oocytes (MI-MII-ICSI, n = 202) underwent an intracytoplasmic sperm injection (ICSI) procedure. Maturation rate, fertilization rate and early embryonic development were compared in both groups. In total, 683 oocytes were collected from 117 ICSI cycles of 117 patients. Among them, 244 (35.7%) were mature MII and 259 (37.9%) were MI after the denudation process. Of those 259 MI oocytes, 202 (77.9%) progressed to MII oocytes after an incubation period of 18–24 h. The maturation rate was 77.9%. Fertilization rate was found to be significantly higher in the rescued in vitro matured MI oocyte group when compared with the in vivo matured MII oocyte group (41.6% vs 25.8%; P = 0.0006). However, no significant difference was observed in terms of cleavage rates on days 2 and 3 between the groups (P = 0.9126 and P = 0.5031, respectively). There may be unidentified in vivo factors on the oocyte maturation causing low developmental capacity in spite of high fertilization rates in the group of patients with low oocyte maturity rate. Furthermore, studies are needed to determine the appropriate culture characteristics as well as culture period and ICSI timing of these oocytes.

scholarly journals Effects of Epidermal Growth Factor and Myo-Inositol on Nuclear Configuration and Subsequent Embryonic Development of Sheep Oocytes

2021 ◽  
Vol 9 (12) ◽  
pp. 2147-2152
Author(s):  
Omar Mardenli ◽  
Hadi Awad Hassooni ◽  
Mahdi Saleh Mohammad Alkerwi
Keyword(s):  
Growth Factor ◽  
Epidermal Growth Factor ◽  
Culture Media ◽  
Calf Serum ◽  
Germinal Vesicle ◽  
Cellular Level ◽  
Biological Interactions ◽  
Epidermal Growth ◽  
Germinal Vesicle Break Down ◽  
Metaphase I

Growth factors and vitamin-like substances have great positive importance in most biological interactions in the cellular level. The addition of these elements in the culture media will increase the yield of the resulting embryos and improve quality. We examined the effects of epidermal growth factor (EGF) and myo-inositol (MI) on meiotic maturation and yields of blastocyst of Awassi sheep oocyte across two experiments. The oocytes obtained were subjected into three treatments: A (without EGF nor MI), B (10 ng/ml EGF + 20 mmol/l MI) and C (50 ng/ml EGF +40 mmol/l MI). Oocytes were then cultured in Ham's F-10 medium supplemented with 5% (v: v) fetal calf serum and 40 ng/ml follicle - stimulating hormone. In the first experiment, during the 27-h culture period, the oocytes were assessed for germinal vesicle break down, metaphase-I and metaphase-II stages across three-time intervals (9, 21 and 27-h). Results of the experiment showed that EGF and MI enhanced the rates of germinal vesicle break down phase (1.53%; 27-h interval; lowest value), metaphase-I (33.87%; 21-h interval) and metaphase-II (89.23%; 27-h interval). In the second experiment, the oocytes incubated in treatment B achieved the highest rates of cleavage (81.96%), 2-8 cell (62.35%) and blastocyst (45.09%). It is concluded from the present study that incubating sheep oocytes in culture media containing a cocktail of EGF (10 ng/ml) and MI (20 mmol/l) significantly improves the rates of metaphase-II, fertilization and blastocyst rates.

Cleavage Rate of in vitro Matured Oocytes Fertilized with Conventional Semen in Buffaloes

2021 ◽  
Author(s):  
M.H. Pitroda ◽  
K.P. Khillare ◽  
M.B. Amle ◽  
M.D. Meshram ◽  
A.B. Mali ◽  
...  
Keyword(s):  
Fertilization Rate ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Quick Recovery ◽  
Slaughter House ◽  
Maturation Rate ◽  
Aspiration Technique ◽  
Current Scenario ◽  
In Vitro Embryo Production

Background: In vitro embryo production in buffaloes has gained much importance in this current scenario due to ever increasing population and high demand of milk and meat. Slaughter house derived bubaline ovaries are a cheap and abundant source of cumulus oocyte complexes.Methods: Oocytes from the buffalo ovarian follicles were recovered by aspiration technique as it facilitates quick recovery. Total 155 ovaries were used in the present study. Surface follicles were measured using vernier calliper and categorized into three groups viz. less than 3 mm, 3-5 mm and greater than 5 mm based on follicular diameter and oocytes were processed for IVM, IVF and IVC using conventional non sorted semen.Result: Overall percentage of small, medium and large follicles in the ovaries were recorded as 16.29 ± 0.94%, 8.14±0.60%, 5.35 ± 0.76%, respectively. Overall recovery rate of COCs was 38%. The percentage of these oocytes were 16.74% (A), 15.25% (B), 25.26% (C), 18.33% (D) and 29.87% (E) respectively. Maturation rate of oocytes were 81.96 ± 2.70%. Fertilization rate was 74.98 ± 3.87%, Cleavage rate % was 40.84±2.51% and Blastocyst percentage was 21.57±1.75% respectively. Application of in vitro embryo production technique using slaughter house ovaries can salvage the genetic potential of bubaline species.

P–279 Effect of Ca2+ ionophore A23187 (calcymicin) on fertilization rates, embryo development and pregnancy in human assisted reproduction cycles

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
G Lópe. Ruiz ◽  
C Olmed. Illueca ◽  
M Bare. Gómez ◽  
S Roy. Bolea ◽  
L Aba. d. Velasco ◽  
...  
Keyword(s):  
Assisted Reproduction ◽  
Fertilization Rate ◽  
Control Group ◽  
Ionophore A23187 ◽  
Oocyte Activation ◽  
Fertilization Failure ◽  
Reproductive Outcomes ◽  
Blastocyst Formation ◽  
Fertilization Rates ◽  
Embryo Kinetics

Abstract Study question Does Calcymicin improve reproductive outcomes of ICSI cycles in cases of fertilization failure and/or embryo blockage indications? Summary answer The application of the Calcymicin after ICSI improves reproductive outcomes, especially in cases with clinical indication of fertilization failure. What is known already According to the bibliography, deficiencies in the oocyte activation process frequently lead to failed ICSI cycles, and these can be corrected by increasing initial levels of calcium (Ca2+) in the oocyte using assisted oocyte activation techniques (AOA), such as the use of Ca2+ ionophores. Ca2+ Ionophores have been shown to trigger an initial Ca2+ spike in the ooplasm that activates Ca2+/Calmodulin dependent protein kinase II, which initiates the cascade of cellular events leading to oocyte activation. Previous results suggest that Ca2+ ionophore treatment can give live offspring after failed ICSI cycles. Study design, size, duration 270 oocytes collected from 17 patients who presented cycles with low fertilization rates and/or embryo blockage or poor quality embryos (according to ASEBIR’s embryo classification criteria) were retrospectively analyzed. Oocytes were divided into two groups, a control group that underwent conventional IVF/ICSI and another group that underwent an ICSI cycle with AOA. Study groups were defined according to clinical indications and subgroups according to AOA or control. All data were collected from 2017 until 2020. Participants/materials, setting, methods Among the 270 oocytes of the study sample, 142 belonged to the control group and 128 belonged to the AOA group. The AOA group oocytes were activated for 15 minutes immediately after ICSI using a prepared solution containing the Ca2+ ionophore A23187, CultActive© (Gynemed, Germany). Fertilization rate and type, blastocyst formation rate, blastocyst quality, embryo kinetics, and pregnancy rates were analyzed, all of them were compared to FIV/ICSI cycles without oocyte activation (control group). Main results and the role of chance In the analyses of the whole sample of oocytes, the AOA treatment gave a fertilization rate of 72.5%, which was significantly higher compared to 53.8% of the control cycles (p = 0.002). Good quality blastocysts and pregnancy rates were also significantly higher than the control (p = 0.01). In the group with an indication of fertilization failure, a significantly higher fertilization rate was recorded compared to the control (65% and 33%, respectively). A higher rate of abnormal embryos with three pronuclei was also found compared to the control (p < 0.001). There were no significant differences in blastocyst formation rates, quality, or embryo kinetics (p > 0.05). In the group with an indication of embryo blockage/poor embryo quality, a significantly higher rate of good quality blastocysts and lower blastulation time were recorded compared to the control (p < 0.05). Limitations, reasons for caution The safety of the AOA technique with Ca2+ ionophore has not been fully demonstrated. In our study, none of the newborns had malformations, and gestational weeks and birth weights were normal. However, further studies on the safety of this technique are needed to implement it routinely in human reproduction clinics. Wider implications of the findings: According to these findings, an increase in the initial levels of calcium in the oocyte through the application of the Ca2+ ionophore A23187 after ICSI improves the results of failed assisted reproduction cycles, especially in the case of those diagnosed with fertility failure, which is a clear indication for AOA. Trial registration number Not applicable

scholarly journals Inseminating dose and water volume applied to the artificial fertilization of Steindachneridion parahybae (Steindachner, 1877) (Siluriformes: Pimelodidae): Brazilian endangered fish

2016 ◽  
Vol 14 (1) ◽  
Author(s):  
Eduardo Antônio Sanches ◽  
Danilo Caneppele ◽  
Renan Yoshiharu Okawara ◽  
Danielle Zanerato Damasceno ◽  
Robie Allan Bombardelli ◽  
...  
Keyword(s):  
Significant Interaction ◽  
Water Volume ◽  
Experimental Unit ◽  
Vitro Fertilization ◽  
Endangered Fish ◽  
Fertilization Rates ◽  
Randomized Design ◽  
Artificial Fertilization ◽  
Hatching Rates

Abstract The Steindachneridion parahybae is an endangered catfish from Brazil and strategies applied for gametes optimization are necessary. The aim of this study was to assess inseminating doses and water volume upon the fertilization, hatching rates and percentage of normal larvae in S. parahybae . Was used a randomized design in factorial scheme (4×4) with four inseminating doses: 1.0×104, 1.0×105, 1.0×106, 1.0×107spermatozoa oocyte-1 and four volumes of water: 1, 35, 65 and 95mL of water g-1 of oocytes. The combination of doses and volumes were performed in triplicates (n=48). Each incubator (1.5L of useful volume) with 1g of oocytes was considered as an experimental unit. Significant interaction between inseminating doses and volumes of water to the values of the fertilization rates and quadratic effect of doses and volume for the values of hatching rates were observed. The doses and volumes did not influence the percentage of normal larvae (87.70±5.06%). It is recommended the use of 5.5×106 spermatozoa oocyte-1 and 1mL of water g-1 of oocytes during in vitro fertilization procedure. These results allowed us to develop new biotechnological strategies applied to the conservation of S. parahybae .

scholarly journals Cytoplasmic Transfer Improves Human Egg Fertilization and Embryo Quality: an Evaluation of Sibling Oocytes in Women with Low Oocyte Quality

2020 ◽  
Author(s):  
Ales Sobek ◽  
Emil Tkadlec ◽  
Eva Klaskova ◽  
Martin Prochazka
Keyword(s):  
Embryo Quality ◽  
Oocyte Quality ◽  
Fertilization Rate ◽  
Early Embryo ◽  
Vitro Fertilization ◽  
Sibling Oocytes ◽  
Fertilization Rates ◽  
Group B ◽  
Cytoplasmic Transfer

Abstract The aim of this study was to evaluate if cytoplasmic transfer can improve fertilization and embryo quality of women with oocytes of low quality. During ICSI, 10–15% of the cytoplasm from a fresh or frozen young donor oocyte was added to the recipient oocyte. According to the embryo quality, we defined group A as patients in which the best embryo was evident after cytoplasmic transfer and group B as patients in which the best embryo was evident after a simple ICSI. We investigated in the period of 2002–2018, 125 in vitro fertilization cycles involving 1011 fertilized oocytes. Five hundred fifty-seven sibling oocytes were fertilized using ICSI only and 454 oocytes with cytoplasmic transfer. Fertilization rates of oocytes were 67.2% in the cytoplasmic transfer and 53.5% in the ICSI groups (P < 0.001). A reduction in fertilization rate was observed with increased women age in the ICSI but not in the cytoplasmic transfer groups. The best embryo quality was found after cytoplasmic transfer in 78 cycles (62.4%) and without cytoplasmic transfer in 40 cycles (32%, P < 0.001). No significant differences were detected between the age, hormonal levels, dose of stimulation drugs, number of transferred embryos, pregnancy rate and abortion rate between A and B groups. Cytoplasmic transfer improves fertilization rates and early embryo development in humans with low oocyte quality. All 28 children resulting from cytoplasmic transfer are healthy.

scholarly journals Prospective Randomized Study on the Influence of Myoinositol in PCOS Women Undergoing IVF in the Improvement of Oocyte Quality, Fertilization Rate, and Embryo Quality

2016 ◽  
Vol 2016 ◽  
pp. 1-5 ◽  
Author(s):  
Bernd Lesoine ◽  
Pedro-Antonio Regidor
Keyword(s):  
Placebo Group ◽  
Folic Acid ◽  
Embryo Quality ◽  
Randomized Study ◽  
Oocyte Quality ◽  
Fertilization Rate ◽  
Clear Trend ◽  
Fertilization Rates ◽  
Group B

Polycystic ovarian syndrome (PCOS) is one of the pathological factors involved in the failure of in vitro fertilization (IvF). The aim of the present study was to investigate if the combination of myoinositol + folic acid was able to improve the oocyte quality, the ratio between follicles and retrieved oocytes, the fertilization rate, and the embryo quality in PCOS patients undergoing IvF treatments. 29 patients with PCOS underwent IvF protocols for infertility treatment and were randomized prospectively into two groups. Group A (placebo) with 15 patients and group B (4000 mg myoinositol + 400 μg folic acid per day) with 14 patients. The patients of group B used for two months myoinositol + folic acid before starting the IvF protocol and data were obtained concerning number of follicles, number of oocytes, quality of oocytes, fertilization rates, and embryo quality in both groups. The ratio follicle/retrieved oocyte was better in the myoinositol group (= group B). Out of the 233 oocytes collected in the myoinositol group 136 were fertilized, whereas only 128 out of 300 oocytes in the placebo group were fertilized. More metaphase II and I oocytes were retrieved in relation to the total amount of oocytes in the myoinositol. More embryos of grade I quality were obtained in the myoinositol. The duration of stimulation was 9,7 days (±3,3) in the myoinositol group and 11,2 (±1,8) days in the placebo group and the number of used FSH units was lower in the myoinositol group: 1750 FSH units (mean) versus 1850 units (mean). Our evidence suggests that myoinositol therapy in women with PCOS results in better fertilization rates and a clear trend to a better embryo quality. As the number of retrieved oocytes was smaller in the myoinositol group, the risk of hyper stimulation syndrome can be reduced in these patients.

Towards the enhancement of pregnancy rate: The effect of insemination time on sperm transport, fertilization rate and embryo quality in dairy cattle

2001 ◽  
Vol 26 (1) ◽  
pp. 161-174 ◽  
Author(s):  
J.C. Dalton ◽  
S. Nadir ◽  
J. Bame ◽  
M. Noftsinger ◽  
R.G. Saacke
Keyword(s):  
Dairy Cattle ◽  
Pregnancy Rate ◽  
Embryo Quality ◽  
Fertilization Rate ◽  
Pregnancy Rates ◽  
Sperm Number ◽  
Sperm Transport ◽  
Fertilization Rates ◽  
Natural Service

AbstractTo further identify factors which influence pregnancy rates, three experiments were conducted to determine the effect of insemination time on sperm transport, fertilization rate, and embryo quality. All cows were continuously monitored for behavioural oestrus by HeatWatch®, and received AI at heat onset (0 h after the first standing event), 12 h after onset, or received natural service at 0 hfrom one of three bulls (Exp. 1). In Exp. 2, cows received AI at 0 h, 12 h, or 24 h after the first standing event. On d 6 after insemination 115 embryos(ova) (Exp. 1) and 117 embryos(ova) (Exp. 2) were recovered from single-ovulating cows. For Exp. 1, median accessory sperm values were: 1 (0 h), 10 (12 h), 27 (natural service O h) (P < 0.05). For Exp. 2, median accessory sperm values were: 1 (0 h), 2 (12 h), 4 (24 h) (P < 0.05). Fertilization rates were: 67% (0 h), 79% (12 h), 98% (natural service O h) (P < 0.05)(Exp. 1); and did not differ in Exp. 2. Embryo quality was not different in Exp. 1. In Exp. 2, percentages of excellent and good fair and poor, and degenerate embryos were: 77, 15, 8 (0 h), 52, 38, 10 (12 h), 47, 19, 34 (24 h) (P < 0.05). In Exp. 3, 30 cows were superovulated and were inseminated once at either 0 h, 12 h, or 24 h after the first standing event. On d 6 after insemination, 529 embryos(ova) were recovered. Fertilization rates were: 29% (0 h); 60% (12 h); 81% (24 h)(P < 0.01). Percentages of embryos with accessory sperm were: 5 (0 h); 8 (12 h); and 41(24 h) (P < 0.01). Embryo quality was not affected by time of AI. We conclude that the time of insemination affects: 1) sperm transport as measured by median accessory sperm number (Exp. 1 and 2) and the percentage of embryos with accessory sperm (Exp. 3); 2) fertilization rate (Exp. 1 and 3); and embryo quality (Exp. 2).

Overheating is detrimental to meiotic spindles within in vitro matured human oocytes

Zygote ◽  
2004 ◽  
Vol 12 (1) ◽  
pp. 65-70 ◽  
Author(s):  
Xiao-Fang Sun ◽  
Wei-Hua Wang ◽  
David L. Keefe
Keyword(s):  
High Temperature ◽  
Light Microscope ◽  
Polarized Light ◽  
Germinal Vesicle ◽  
Meiotic Spindle ◽  
Human Oocytes ◽  
Living State ◽  
Meiotic Spindles ◽  
Metaphase I

The present study was designed to examine the effects of overheating on meiotic spindle morphology within in vitro matured human oocytes using a polarized light microscope (Polscope). Immature human oocytes at either germinal vesicle or metaphase I stage were cultured in vitro for 24–36 h until they reached metaphase II (M-II) stage. After maturation, oocytes at M-II stage were imaged in the living state with the Polscope at 37, 38, 39 and 40 °C for up to 20 min. After heating, oocytes were returned to 37 °C and then imaged for another 20 min at 37 °C. The microtubules in the spindles were quantified by their maximum retardance, which represents the amount of microtubules. Spindles were intact at 37 °C during 40 min of examination and their maximum retardance (1.72–1.79) did not change significantly during imaging. More microtubules were formed in the spindles heated to 38 °C and the maximum retardance was increased from 1.77 before heating to 1.95 at 20 min after heating. By contrast, spindles started to disassemble when the temperature was increased to 39 °C for 10 min (maximum retardance was reduced from 1.76 to 1.65) or 40 °C for 1 min (maximum retardance was reduced from 1.75 to 1.5). At the end of heating (20 min), fewer microtubules were present in the spindles and the maximum retardance was reduced to 0.8 and 0.78 in the oocytes heated to 39 °C and 40 °C, respectively. Heating to 40 °C also induced spindles to relocate in the cytoplasm in some oocytes. After the temperature was returned to 37 °C, microtubules were repolymerized to form spindles, but the spindles were not reconstituted completely compared with the spindles imaged before heating. These results indicate that spindles in human eggs are sensitive to high temperature. Moreover, maintenance of an in vitro manipulation temperature of 37 °C is crucial for normal spindle morphology.

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