PSX-A-23 Late-Breaking: Supplementation of Nerve Growth Factor (β-NGF) in the maturation medium and efficiency of in vitro embryo production in cattle

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 365-365
Author(s):  
Lucas Gonçalves ◽  
Muller C Martins ◽  
Natalia Arle ◽  
Rafaela T Torres ◽  
Luisa Migilo ◽  
...  
Keyword(s):  
Nerve Growth Factor ◽  
Growth Factor ◽  
Oocyte Maturation ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Nerve Growth ◽  
Maturation Medium ◽  
In Vitro Embryo Production ◽  
Blastocyst Rate

Abstract The aim of this study was to evaluate the supplementation of Nerve Growth Factor (β-NGF) in the maturation medium in in vitro embryo production routines. Antral follicles were aspirated from ovaries of cows obtained from slaughterhouses and then oocytes were selected for quality (grades I and II) for in vitro maturation and subjected to 4 successive in vitro embryo production routines (IVEP). Supplementation of 100 ng of β-NGF was performed in the oocyte maturation medium 22 hours before in vitro fertilization. 48 hours after fertilization of the oocytes, an analysis was made of their cleavage rate by counting blastomeres with the aid of a stereoscopic microscope (cleavage rate = number of embryos / number of initial oocytes). Seven days after fertilization, the blastocyst rate was determined through the relation to the number of oocytes that started cleavage and reached this stage of development (blastocyst rate = number of blastocyst / number of oocytes that started cleavage). To verify the existence of a difference between the supplemented and the non-supplemented groups, the paired T test was applied, using the Excel / Action software (Microsoft). In vitro embryo production routines supplemented with β-NGF in the maturation medium had, on average, a higher cleavage rate (P = 0.0072) and a higher blastocyst rate (P = 0.0033) compared to non-supplemented routines with β-NGF. In this study was demonstrated that Nerve Growth Factor supplementation in the maturation medium improves the efficiency of in vitro embryo production in cattle, and this protein has a probable action in the oocyte maturation process.

scholarly journals Effect of deslorelin acetate treatment in oocyte recovery and in vitro embryo production in domestic cats

2016 ◽  
Vol 19 (10) ◽  
pp. 1091-1095
Author(s):  
Camila Louise Ackermann ◽  
Eduardo Trevisol ◽  
Leticia Ferrari Crocomo ◽  
Tatiana da Silva Rascado ◽  
Rodrigo Volpato ◽  
...  
Keyword(s):  
Treated Group ◽  
Control Group ◽  
Domestic Cats ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Oocyte Recovery ◽  
Significant Difference ◽  
In Vitro Embryo Production ◽  
Blastocyst Rate

Objectives The present study investigated the effect of contraceptive treatment with deslorelin acetate on in vitro embryo production and oocyte recovery in domestic queens. Methods Twenty-one mature domestic cats were used. Eleven queens (treated group) and one tom were kept in an experimental cattery, and 10 queens were privately owned (control group). When in interestrus or diestrus (day 0) a deslorelin acetate implant (Suprelorin, 4.7 mg/animal) was inserted into the subcutaneous tissue of the interscapular region in all queens in the treated group. After 6 months of treatment, all animals were ovariohysterectomized, and the ovaries were used for in vitro embryo production. Percentage of cleavage was determined 18 h after oocyte insemination and blastocyst formation was assessed on the eighth day of culture. The rate of cumulus-oocyte complexes (COCs) recovery was analyzed by an unpaired t-test. The cleavage and blastocyst rates were expressed as percentages and analyzed by Fisher’s exact test. All analyses were performed using GraphPad Prism v5.0, with P <0.05 set as the level of significance. Results In the treated group, we recovered 8.3 ± 1.15 grade I COCs per queen; the cleavage rate was 60% and the blastocyst rate was 36%. In the control group, we recovered 18.4 ± 3.21 grade I COCs per queen; the cleavage rate was 55.97% and the blastocyst rate was 34%. Forty percent of treated females did not produce any blastocysts. In the treated group, we observed a significant decrease in COC recovery. Although there was no significant difference in cleavage and blastocyst rates between groups, 40% of treated females did not produce any blastocysts. Conclusions Recovery of grade I COCs is negatively affected by deslorelin treatment in domestic cats. Regarding embryo production, new studies are still necessary to evaluate the success of this technique owing to the individual effect caused by deslorelin acetate.

177 PERFORMANCE OF Gyr PREPUBERTAL HEIFERS IN IN VITRO EMBRYO PRODUCTION

2016 ◽  
Vol 28 (2) ◽  
pp. 219
Author(s):  
P. M. S. Rosa ◽  
A. J. R. Camargo ◽  
R. V. Serapião ◽  
L. S. A. Camargo ◽  
C. S. Oliveira
Keyword(s):  
Cleavage Rate ◽  
Embryo Production ◽  
Exact Test ◽  
Follicular Wave ◽  
Lactating Cows ◽  
In Vitro Embryo Production ◽  
Blastocyst Rate ◽  
Oocyte Donors ◽  
La Jolla

Bovine in vitro embryo production is highly relevant for dairy systems in Brazil, and Gyr dams are commonly used as oocyte donors. The aim of this study was to evaluate the use of prepubertal Gyr heifers as oocyte donors, an alternative to anticipate reproduction of those animals. For that, 11 Gyr [4 prepubertal (PP) donors and 7 adult cows © donors] were used in ovum pickup (OPU) sessions. The PP cows presented an average of 282.5 kg and 26.75 months, and had never displayed oestrous. Non-lactating cows presenting an average of 492 kg and 136 months were selected for C. Five replicates were performed, totaling 27 OPU sessions (C-17, PP-10) and 2–3 sessions per animal. Follicular wave was synchronised by aspiration of follicles larger than 8 mm 96 h before OPU. Cumulus-oocyte complexes (COC) were classified accordingly to their quality in viable (G1, G2, and G3) or non-viable (G4). Viable oocytes were matured and fertilized, and the presumptive zygotes were cultured in SOF medium at 38.5°C and 5% CO2 in air. Cleavage rate was assessed 48 to 72 h post-insemination (hpi) and blastocyst rate at 168 hpi. Mean number of structures was analysed by t-test, and percentage of viable, G1, G2, G3, G4, cleavage, and blastocyst rates were compared among groups by Fisher’s exact test (GraphPadInstat, La Jolla, CA, USA; P = 0.05). Results are followed by standard error values. All procedures were approved by a local ethics committee. We found that despite higher (P < 0.05) numbers for both viable oocytes (PP: 15 ± 2.6; C: 6.11 ± 0.76) and total oocytes (PP: 23.70 ± 2.83; C: 8.82 ± 1.19) in the PP group, the rate of viable oocytes was similar (P > 0.05) among PP and C groups (PP: 61.5 ± 6.51%, C: 66.79 ± 3.79%). Mean numbers of G1, G2, G3, and G4 oocytes were higher (P < 0.05) in the PP group (G1 = 7.1 ± 1.18; G2 = 4.9 ± 1.74; G3 = 3.9 ± 1.09; G4 = 7.8 ± 1.38) than in the C group (G1 = 2.70 ± 0.740; G2 = 2.47 ± 0.44; G3 = 1.11 ± 0.31; G4 = 2.52 ± 0.39). However, the proportion was similar (P > 0.05) among PP and C groups (PP: G1 = 29.5 ± 4.21%; G2 = 19.5 ± 2.85%; G3 = 15.9 ± 13.5%; G4 = 35.1 ± 6.33%; and C: G1 = 27.24 ± 4.44%; G2 = 29.60 ± 5.08%; G3 = 12.34 ± 3.01%, G4 = 30.79 ± 4.93%). Cleavage rate (PP: 91.3 ± 17.94%; C: 74.09 ± 4.65%), mean blastocyst number per OPU session (PP: 3.3 ± 1.29; C: 1.76 ± 0.28), and blastocyst rate (PP: 19.74 ± 7.40%; C: 27.03% ± 4.07%) were similar (P > 0.05) among groups. We conclude that prepubertal heifers presented increased numbers of viable oocytes per OPU session, but blastocyst yield was similar to adult cows. This data suggests that prepubertal Gyr heifers can be used as oocyte donors. Support from FAPERJ and Embrapa is acknowledged.

scholarly journals Exposure to follicular fluid during oocyte maturation and oviductal fluid during post-maturation does not improve in vitro embryo production in the horse

Zygote ◽  
2017 ◽  
Vol 25 (5) ◽  
pp. 612-630 ◽  
Author(s):  
Cécile Douet ◽  
Olivia Parodi ◽  
Nicola Antonio Martino ◽  
Giovanni Michele Lacalandra ◽  
Michele Nicassio ◽  
...  
Keyword(s):  
Oocyte Maturation ◽  
Follicular Fluid ◽  
Culture Conditions ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Development Rates ◽  
Maturation Medium ◽  
In Vitro Embryo Production ◽  
Oviductal Fluid

SummaryMost wild equids and many domestic horse breeds are at risk of extinction, so there is an urgent need for genome resource banking. Embryos cryopreservation allows the preservation of genetics from male and female and is the fastest method to restore a breed. In the equine, embryo production in vitro would allow the production of several embryos per cycle. Intracytoplasmic sperm injection (ICSI) is used to generate horse embryos, but it requires expensive equipment and expertise in micromanipulation, and blastocyst development rates remain low. No conventional in vitro fertilization (IVF) technique for equine embryo production is available. The development of culture conditions able to mimic the maturation of the oocyte in preovulatory follicular fluid (pFF) and the post-maturation in oviductal fluid (OF) may improve embryo production in vitro. Our aim was to analyse the effect of in vitro maturation in pFF and incubation in OF on in vitro maturation of equine oocytes, fertilization using conventional IVF or ICSI, and embryo development after culture in synthetic oviductal fluid (SOF) or DMEM-F12. Oocytes collected from slaughtered mares or by ovum pick up were matured in vitro in pFF or semi-synthetic maturation medium (MM). The in vitro maturation, fertilization and development rates were not statistically different between pFF and MM. After in vitro maturation, oocytes were incubated with or without OF. Post-maturation in OF did not significantly improve the fertilization and development rates. Thus, in our study, exposure to physiological fluids for oocyte maturation and post-maturation does not improve in vitro embryo production in the horse.

111 EFFECT OF ETHYLENE GLYCOL IN VITRIFICATION MEDIUM ON BOVINE OOCYTE ACTIVATION AND IN VITRO EMBRYO PRODUCTION

2010 ◽  
Vol 22 (1) ◽  
pp. 214
Author(s):  
G. L. Rios ◽  
G. G. Kaiser ◽  
N. C. Mucci ◽  
R. H. Alberio
Keyword(s):  
Ethylene Glycol ◽  
Control Group ◽  
Oocyte Activation ◽  
Bovine Oocyte ◽  
Cleavage Rate ◽  
Hoechst 33342 ◽  
Embryo Production ◽  
Maturation Medium ◽  
In Vitro Embryo Production

In this study the effect of increasing ethylene glycol (EG) concentrations in the vitrification media (VM) and its relation with oocyte activation and in vitro embryo production were evaluated. Cumulus-oocytes complexes (COC) were matured in vitro for 22 h and then partially denuded by gently pipetting with hyaluronidase, and randomly assigned to 4 treatments: T1 = control group; T2 = COC exposed to 10% EG + 10% DMSO (VM1), 20% EG + 20% DMSO (VM2); T3 = 15% EG + 5% DMSO (VM1), 30% EG + 10% DMSO (VM2); T4 = 20% EG + 0% DMSO (VM1), 40% EG + 0% DMSO (VM2). Exposition to VM1 and VM2 lasted 3 min and 30 s, respectively. After treatment COC were incubated in maturation medium up to 24 h. In Experiment 1 COC were cultured for 24 h in fertilization TALP supplemented with 50 μg mL-1 of heparin, then completely denuded and cultured 24 h in CR1-aa. After this, oocytes were stained with bisbenzimide (Hoechst 33342) and the number of nuclei and cells were recorded. Structures presenting 2 cells or 2 nuclei were considered as activated oocytes. In Experiment 2 matured COC exposed to cryoprotectants as in Experiment 1 were fertilized and cultured for 6 days as previously described (Mucci et al. 2006). Variables analyzed included oocyte activation, cleavage rate at 48 h, and percentage of viable embryos at Day 7. Data were analyzed by PROC GENMOD (SAS Institute Inc., Cary, NC, USA). Activated oocytes percentage did not differ between EG concentrations in VM and were higher (T2, 24.7%, n = 101; T3, 25.0%, n = 96; T4, 30.2%, n = 119) compared with controls (T1, 9.8%, n = 61). In Experiment 2 no differences were found in cleavage rates (T1, 81.9%, n = 68; T2, 87%, n = 67; T3, 85.9%, n = 61; T4, 84.2%, n = 64) and Day 7 percentage of viable embryos (T1, 34.9%, n = 29; T2, 28.6%, n = 22; T3, 26.8%, n = 19; T4, 27.6%, n = 21) in treated COC. The exposition of COC to cryoprotectants per se could trigger oocyte activation in the range of 10 to 40%. Thanks toAdriana Cano (Instituto Nacional de Tecnología Agropecuaria) for contributions in statistical analysis.

67 SUPPLEMENTATION WITH EPIDERMAL GROWTH FACTOR AND GLIAL CELL-DERIVED NEUROTROPIC FACTOR DURING PORCINE OOCYTE MATURATION STIMULATES BLASTOCYST HATCHING RATE AND QUALITY

2012 ◽  
Vol 24 (1) ◽  
pp. 145
Author(s):  
M. Vafaye Valleh ◽  
M. Rasmussen ◽  
O. Oestrup ◽  
M. Schmidt ◽  
P. Hyttel
Keyword(s):  
Growth Factors ◽  
Growth Factor ◽  
Oocyte Maturation ◽  
Cleavage Rate ◽  
Gene Expression Analyses ◽  
Porcine Oocyte ◽  
Epidermal Growth ◽  
Blastocyst Rate ◽  
Neurotropic Factor

The growth factors glial cell-derived neurotropic factor (GDNF) and epidermal growth factor (EGF) have previously been shown to elicit a functional effect on oocyte competence and embryo development; however, their effects on embryo quality remain elusive. The objective of the present study was to investigate whether supplementation with EGF, GDNF, or a combination of the 2 growth factors during in vitro porcine oocyte maturation has a beneficial effect on the rate of embryonic development, cell number and relative expression of genes related to apoptosis (anti-apoptotic: BCLX; pro-apoptotic: BAK, CASPASE), cellular stress (HSP70), mitochondrial function (TFAM) and pluripotency (TERT). Cumulus-oocyte complexes (COC) aspirated from slaughterhouse ovaries were matured for 42 h in maturation medium supplemented with either 50 μg mL–1 of EGF (n = 3398), 50 μg mL–1 of GDNF (n = 1870), or a combination of both growth factors [EGF (50 μg mL–1) + GDNF (50 μg mL–1); EGF+GDNF-group; n = 1993). Subsequently, the oocytes were inseminated, the cleavage rate scored and the resultant embryos cultured in vitro for 7 days. The experiments were replicated a total of 7 times and zona-enclosed blastocysts were collected for Hoechst staining (cell counting) and for gene expression analyses. For gene expression analyses, total RNA was isolated from pools of embryos (n = 10), reverse transcribed into cDNA and subjected to comparative real-time PCR using the delta-delta Ct method (2–ΔΔCT) and statistical method with a significance level of P < 0.05. The blastocyst rate was significantly lowest in the GDNF group (P < 0.05), whereas the hatched blastocyst rate (P < 0.05) was highest in the EGF+GDNF-group (Table 1). The GDNF-group displayed 2-fold up-regulation of BAK compared with the EGF group (P < 0.05), 1.75-fold up-regulation of TFAM compared with the EGF+GDNF group (P < 0.05) and 2.4-fold up-regulation of CASPASE compared with the EGF+GDNF group (P < 0.05). In contrast, an 11-fold decrease of TERT compared with the EGF+GDNF and EGF group was observed (P < 0.05). The EGF+GDNF-group displayed 1.16 and 1.56-fold up-regulation of BCLX compared with the EGF and GDNF (P < 0.05) groups, respectively and a 1.1-fold up-regulation of TERT compared with the EGF group; in contrast, there was a 1.35 and 1.25 fold down-regulation of CASPASE and TFAM compared with the EGF group, respectively. The EGF-group displayed the lowest (P < 0.05) expression level of BAK. In conclusion, compared among treatments, supplementation with a combination of EGF and GDNF during porcine in vitro oocyte maturation resulted in an increased rate of hatched blastocysts and increased expression of anti-apoptotic (BCLX) and pluripotency related (TERT) genes, whereas it resulted in decreased expression of the pro-apoptotic gene (CASPASE). The present study may help to improve the culture conditions, which are important for optimizing blastocyst rates and quality. Table 1.Effect of using growth factor in in vitro maturation on the cleavage rate and blastocyst yield (mean ± standard error of the means) in vitro

122 A New Maturation Medium Improves Porcine Embryo Production In Vitro

2018 ◽  
Vol 30 (1) ◽  
pp. 200 ◽  
Author(s):  
A. Lucas-Hahn ◽  
B. Petersen ◽  
M. Nowak-Imialek ◽  
U. Baulain ◽  
R. Becker ◽  
...  
Keyword(s):  
Growth Factor ◽  
Cell Number ◽  
Developmental Competence ◽  
Gestation Length ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Vitro Fertilization ◽  
Maturation Medium

Recently (Spate et al. 2017 Reprod. Fertil. Dev. 29, 150), a new medium [TCM-199 supplemented with hCG 10 IU, pregnant mare serum gonadotropin (PMSG) 10 IU mL−1, fibroblast growth factor (FGF) 40 ng mL−1, leukemia inhibitory factor (LIF) 2000 U mL−1, IGF-1 20 ng mL−1, epidermal growth factor (EGF) 10 ng mL−1], termed FLI medium, was demonstrated to improve porcine oocyte maturation in vitro. The effects on embryo development and quality have not yet been investigated. The purpose of the present study was to compare the FLI medium in porcine in vitro embryo production (IVP) with our standard maturation medium (DMEM supplemented with 10 IU mL−1 PMSG and hCG, 50 ng mL−1 EGF, 100 ng mL−1 IGF1, and 5 ng mL−1 FGF). Briefly, gilt oocytes were collected via aspiration of follicles from abattoir ovaries and matured for 44 h in either FLI or standard DMEM medium at 39°C, 5% CO2 in humidified air. In vitro fertilization was performed with freshly ejaculated sperm (250,000 mL−1) of a multi-transgenic boar (GGTA1-KO/hCD46/hCD55/hCD59/hHO-1/hA20) by co-incubation with the matured oocytes in PGMTac4 medium for 4 h. Zygotes were washed twice and then cultured for 6 days in PZM3 medium. Development to the blastocyst stage was recorded at Day 6 of culture. Blastocysts were fixed and Hoechst33342 stained for counting the nuclei. Each of the experiments was repeated 3 times. In a second step, Day 5 blastocysts derived from the FLI medium were transferred to synchronized pubertal gilts to test the in vivo developmental competence of the IVF embryos. Maturation of oocytes in FLI medium resulted in a significantly higher blastocyst rate (49.3 vs. 13.5; P ≤ 0.001, Chi-squared test) and nuclei number (41.3 ± 12.2 vs. 35.3 ± 10.8; P ≤ 0.001, one-way ANOVA) compared with the standard medium, whereas the cleavage rate was not affected. Transfer of Day 5 blastocysts (average 35 embryos/recipient) derived from the FLI system using 8 recipients resulted in 7 pregnancies (87.5%) as determined by ultrasound scanning on Day 25 of gestation. At the time of writing, one recipient had delivered 5 healthy piglets after a gestation length of 114 days. Results indicate that the FLI medium significantly improves blastocyst rates and the cell number of the resulting blastocysts (Table 1) and yields pig IVF embryos with a high developmental capacity in vivo. By producing high-quality porcine embryos, this FLI-based IVF system provides an efficient method to modify the porcine genome by cytoplasmic microinjection of CRISPR/Cas molecules into IVF-derived zygotes. Table 1.Results of maturation of oocytes in FLI medium compared with DMEM

178 FIRST COMMERCIAL CATTLE IN VITRO EMBRYO PRODUCTION AND PREGNANCY RATES OF BOTH FRESH AND FROZEN IN VITRO EMBRYOS IN THE NORTH COAST OF PERU

2016 ◽  
Vol 28 (2) ◽  
pp. 220
Author(s):  
H. W. Vivanco-Mackie ◽  
R. D. Navarro ◽  
M. D. P. Salazar ◽  
E. A. Aguirre ◽  
G. B. Saldaña ◽  
...  
Keyword(s):  
Growth Factor ◽  
Pregnancy Rate ◽  
Artificial Insemination ◽  
North Coast ◽  
Embryo Production ◽  
The North ◽  
In Vitro Embryo Production ◽  
Blastocyst Rate ◽  
North Coast Of Peru

The objective of the study was to determine the pregnancy rate of fresh and frozen-thawed in vitro produced embryos transferred into recipients in the north coast of Peru. Artificial insemination results of frozen-thawed semen inseminated to cows in the same herd and season (summer) where the embryo transfers were performed was evaluated as control. For the in vitro embryo production, the rate obtained was 374 oocytes from 21 ovum pickup sessions (15.24 ± 8.91 oocytes/session). Of these oocytes, 246 were matured in bicarbonate buffered TCM-199 supplemented with 10% heat inactivated FCS as well as epidermal growth factor (EGF), insulin-like growth factor-1 (IGF-1), FSH, LH, oestradiol, and cysteamine for 24 h of incubation at 38.5°C, 5% CO2, and 90% humidity. The oocytes selected post-maturation were fertilised with the frozen-thawed sperm that was subjected post-thawing to Percoll gradient (90 and 45% Percoll), centrifugated, and resuspended in a TALP-IVF medium supplemented with 20 μM D-penicillamine, 10 μM hypotaurine, and 1 μM epinephrine. The oocytes were then inseminated with a concentration of 1.5 million sperm mL–1 in TALP IVF fertilization medium and incubated for 24 h at 38.5°C, 5% CO2, and 90% humidity. Subsequently, the presumptive zygotes were transferred to medium of 50 μL drops of SOFaa supplemented with 5% heat-inactivated FCS which was later replaced by SOFaa and 1% heat-inactivated FCS on Day 5 after fertilization. The embryos were inspected and graded on Days 7 and 8 post-fertilization and incubation at 38.5°C, 5% CO2 and 90% humidity. The blastocyst rate was evaluated on Day 7 post-fertilization. The blastocyst rate was 25.3% (21/83) and 4.19 ± 3.37 embryos per ovum pickup session were obtained. The embryo freezing media contained 1.5 M ethylene glycol as a cryo-protectant, and the method of thawing the embryo was the direct method (1 step). The pregnancy rate was compared by chi-squared analysis. The pregnancy rate for artificial insemination was 23.9% (1103/4612), and the pregnancy rate of fresh and frozen-thawed in in vitro embryos was 30% (13/43) and 20% (8/40), respectively (P > 0.05). Overall the pregnancy rates in the herd were relatively low, probably due the high environmental temperature during the season when the embryos were transferred and the semen was inseminated. Under those conditions, pregnancy rate was not affected by the use of fresh and frozen-thawed in vitro embryo transfer in comparison to the conventional artificial insemination of frozen semen in the coast north of Peru.

140 A comparison of in vitro embryo production between heifers and lactating Holstein donors without superstimulation

2019 ◽  
Vol 31 (1) ◽  
pp. 195
Author(s):  
E. P. Silva ◽  
M. K. Sermersheim ◽  
P. V. Marchioretto ◽  
R. Della Mea ◽  
L. M. Naves ◽  
...  
Keyword(s):  
Donor Age ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Exact Test ◽  
Lactating Cows ◽  
Cumulus Oocyte Complex ◽  
Sexed Semen ◽  
In Vitro Embryo Production ◽  
Blastocyst Rate

Bovine oocyte donor age seems to have an important role on in vitro embryo production. Therefore, the aim of this study was to compare cumulus-oocyte complex (COC) recovery, cleavage, and blastocyst rates between heifers and lactating Holstein donors. A total of 89 animals (heifers: n=60, 11 to 17 months of age; lactating cows: n=29, 60 to 180 days in milk) were used in this experiment. Ovum pickup (Day −1) was performed on a random day of the oestrous cycle without superstimulation by a single technician. Conventional or sexed semen were used for IVF (Day 0). Cleavage and blastocyst rates were evaluated on Days 3 and 7, respectively. Cleavage was determined when structures had 4 or more cells and blastocyst rate included grades I and II blastocysts. Continuous data were analysed by t-test, and binomial data were analysed by Fischer’s exact test. A total of 1,289 COC were recovered from lactating cows (n=509) and heifers (n=777). Total and viable COC per donor were higher (P&lt;0.05) in lactating cows (total=17.55±13.08; viable=11.24±8.9) than heifers (total=12.95±5.39; viable=7.88±4.08). The percentage of viable COC was similar (P&gt;0.05) in lactating cows (64.04%; 326/509) and heifers (60.87%; 473/777). Cleavage rate was higher (P&lt;0.0001) in lactating cows (58.74%; 299/509) than in heifers (45.05%; 350/427). Cleavage rate was similar (P&gt;0.05) with conventional and sexed semen in heifers (conventional=37.68%; 52/138; sexed=46.63%; 298/639) and cows (conventional=55.05%; 49/89; sexed=59.52%; 250/470). Blastocyst rate was higher (P&lt;0.0001) in lactating cows (19.84%; 101/509) than in heifers (8.49%; 66/777). Conventional semen had a higher (P&lt;0.0001) blastocyst rate (18.11%; 24/138) than sexed semen (6.57%; 42/639) in heifers. However, there was no difference (P&gt;0.05) in blastocyst rate between conventional (24.71%; 22/89) and sexed semen (18.8%; 79/420) in lactating cows. Blastocyst/cleaved structures ratio was higher (P&lt;0.05) in heifers using conventional (46.15%; 24/52) compared with sexed (14.06%; 42/298) semen. However, in lactating cows there was a tendency (P&lt;0.1) for higher blastocyst/cleaved structures ratio using conventional (44.89%; 22/49) compared with sexed semen (31.6%; 79/250). In conclusion, a higher number of total and viable COC were aspirated from Holstein lactating cows than heifers. Furthermore, cleavage and blastocyst rates were higher in lactating cows than in heifers. Conventional semen gave more embryos than did sexed semen in heifers but not in cows. Overall, oocytes from Holstein lactating donors are more suitable for in vitro embryo production than are those from heifer donors.

Neuroprotective and anti-amnestic effects of a combination therapy in a model of photochemical ischemic damage in the prefrontal cortex

2018 ◽  
pp. 39-45
Author(s):  
Ф.М. Шакова ◽  
Т.И. Калинина ◽  
М.В. Гуляев ◽  
Г.А. Романова
Keyword(s):  
Prefrontal Cortex ◽  
Nerve Growth Factor ◽  
Growth Factor ◽  
Combination Therapy ◽  
Neuroprotective Effect ◽  
Regulatory Protein ◽  
Human Growth ◽  
Neuroprotective Effects ◽  
Nerve Growth

Цель исследования - изучение влияния комбинированной терапии (мутантные молекулы эритропоэтина (EPO) и дипептидный миметик фактора роста нервов ГК-2H) на воспроизведение условного рефлекса пассивного избегания (УРПИ) и объем поражения коры мозга у крыс с двусторонним ишемическим повреждением префронтальной коры. Методика. Мутантные молекулы EPO (MЕРО-TR и MЕPО-Fc) с значительно редуцированной эритропоэтической и выраженной цитопротекторной активностью созданы методом генной инженерии. Используемый миметик фактора роста нервов человека, эндогенного регуляторного белка, в экспериментах in vitro проявлял отчетливые нейропротективные свойства. Двустороннюю фокальную ишемию префронтальной коры головного мозга крыс создавали методом фотохимического тромбоза. Выработку и оценку УРПИ проводили по стандартной методике. Объем повреждения мозга оценивался при помощи МРТ. MEPO-TR и MEPO-Fc (50 мкг/кг) вводили интраназально однократно через 1 ч после фототромбоза, ГК-2Н (1 мг/кг) - внутрибрюшинно через 4 ч после фототромбоза и далее в течение 4 послеоперационных суток. Результаты. Выявлено статистически значимое сохранение выработанного до ишемии УРПИ, а также значимое снижение объема повреждения коры при комплексной терапии. Полученные данные свидетельствуют об антиамнестическом и нейропротекторном эффектах примененной комбинированной терапии, которые наиболее отчетливо выражены в дозах: МEPO-Fc (50 мкг/кг) и ГК-2Н (1 мг/кг). Заключение. Подтвержден нейропротекторный эффект и усиление антиамнестического эффекта при сочетанном применении мутантных производных эритропоэтина - MEPO-TR и MEPO-Fc и дипептидного миметика фактора роста нервов человека ГК-2H. The aim of this study was to investigate the effect of combination therapy, including mutant erythropoietin molecules (EPO) and a dipeptide mimetic of the nerve growth factor, GK-2H, on the conditioned passive avoidance (PA) reflex and the volume of injury induced by bilateral ischemia of the prefrontal cortex in rats. Using the method of genetic engineering the mutant molecules of EPO, MERO-TR and MEPO-Fc, with strongly reduced erythropoietic and pronounced cytoprotective activity were created. The used human nerve growth factor mimetic, an endogenous regulatory protein based on the b-bend of loop 4, which is a dimeric substituted dipeptide of bis- (N-monosuccinyl-glycyl-lysine) hexamethylenediamine, GK-2 human (GK-2H), has proven neuroprotective in in vitro experiments. Methods. Bilateral focal ischemic infarction was modeled in the rat prefrontal cortex by photochemically induced thrombosis. The PA test was performed according to a standard method. Volume of brain injury was estimated using MRI. MEPO-TR, and MEPO-Fc (50 mg/kg, intranasally) were administered once, one hour after the injury. GK-2Н (1 mg/kg, i.p.) was injected four hours after the injury and then for next four days. Results. The study showed that the complex therapy provided statistically significant retention of the PA reflex developed prior to ischemia and a significant decrease in the volume of injury. The anti-amnestic and neuroprotective effects of combination therapy were most pronounced at doses of MEPO-Fc 50 mg/kg and GK-2H 1 mg/kg. Conclusion. This study has confirmed the neuroprotective effect and enhancement of the anti-amnestic effect exerted by the combination of mutant erythropoietin derivatives, MEPO-TR and MEPO-Fc, and the dipeptide mimetic of human growth factor GK-2H.

Sign in / Sign up

Export Citation Format

Share Document