159 Effect of Commercial Embryo Holding Medium on Development and Quality of Immature Bovine Oocytes

2018 ◽  
Vol 30 (1) ◽  
pp. 219
Author(s):  
M. Catteeuw ◽  
O. B. Pascottini ◽  
G. Opsomer ◽  
A. Van Soom
Keyword(s):  
Low Cost ◽  
Laboratory Work ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Immature Oocytes ◽  
Developmental Capacity ◽  
In Vitro Embryo Production ◽  
Assess Quality ◽  
Bovine Oocytes

Bovine in vitro embryo production following ovum pick-up (OPU) in the field is hampered due to large time gaps between first and last OPU sessions. As oocytes will start immediate maturation, scheduling further manipulations makes the laboratory work laborious. There is a need for an easy and low-cost method that conserves the oocytes with full developmental capacity and allows scheduling laboratory work. In this regard, a commercial embryo holding medium (EHM; Syngro®; Bioniche Inc., WA, USA) was evaluated for conserving immature oocytes. Bovine immature oocytes (n = 2160) were collected from slaughterhouse animals by follicle aspiration and grouped per 60 and stored in 1 mL of EHM in 1-mL sterile glass osmometer tubes. Then, different temperatures [4°C, room temperature (RT), and 38.5°C] and different storage times (6, 10, 14 h) were assessed. After storage, oocytes were matured in TCM-199 supplemented with epidermal growth factor and gentamycin for 22 h. Fertilization was performed and zygotes (n = 1786) were cultured per 25 in 50 µL of SOF medium supplemented with 0.4% BSA and insulin, transferrin, and selenium. Control groups were included: immature oocytes (n = 1080) were not stored in EHM but immediately matured; fertilized and zygotes (n = 896) were cultured. Further, differential apoptotic staining was performed on a random subgroup of blastocysts to assess quality. Generalized fixed effect models were computed using R studio. Storage for 6 h showed a decrease in cleavage and blastocyst rate at 38.5°C (50 ± 3.9%; 11 ± 1.8%) compared with the control (78 ± 3.0%; 36 ± 2.8%). When increasing storage time, 38.5°C was not included; here, 4°C had a lower cleavage and blastocyst yield (47 ± 2.9%; 20 ± 3.3%) compared with the control (75 ± 2.5%; 41 ± 4.6%). For both 6 and 10 h, storage at RT resulted in similar cleavage (76 ± 3.4%; 74 ± 2.6%) and blastocyst rates (35 ± 2.7%; 40 ± 4.5%) as the control (P > 0.05). However, increasing storage to 14 h at RT decreased cleavage (61 ± 2.8%) and blastocyst yield (26 ± 2.5%) compared with the control (78 ± 2.4%; 39 ± 2.8%; P < 0.05). Evaluating embryo quality in all groups, no significant differences were found for any holding time or temperature of the EHM. To simulate OPU settings, EHM was also tested in 38 small groups of 10 immature oocytes that were subsequently matured, fertilized, and cultured. Based on the previous results, EHM storage was performed for 6 and 10 h at RT. Blastocyst development was not different between RT (19.8 ± 3.5%; 18.8 ± 3.6%) and the control (20.6 ± 3.6%; 18.3 ± 3.4%; P ≥ 0.05). To conclude, a commercial EHM can be used to conserve immature bovine oocytes without losing developmental capacity. Storage is recommended for no longer than 10 h and at RT in EHM. It opens new perspectives for practitioners, because this method is simple and low-cost; moreover, the start of maturation and subsequent in vitro embryo production process can be scheduled to avoid evening or night work at the laboratory.

PSX-A-10 Late-Breaking: Effects of paternal high energy diets on blastocyst development during in vitro embryo production in the bovine

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 363-363
Author(s):  
Dylan B Davis ◽  
Zachary Seekford ◽  
Mackenzie Dickson ◽  
Lucas Gonçalves ◽  
Samir Burato ◽  
...  
Keyword(s):  
High Gain ◽  
High Energy ◽  
Carcass Composition ◽  
Adaptation Period ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Ad Libitum ◽  
In Vitro Embryo Production ◽  
To Receive

Abstract The objective of this study was to evaluate the effect of paternal high energy diets on blastocyst development during in vitro embryo production (IVP). Eight sires were stratified by body weight (initial BW = 946 ± 85 kg) and randomly assigned to the same diet (NEm = 2.10, NEg = 1.44, CP = 14.1%, NDF = 16.6%, DM basis) fed at two different inclusion rates while having ad libitum access to bermudagrass hay (NEm = 1.02, NEg = 0.45, CP = 10.2%, NDF = 71.6). After a 10-d adaptation period, sires were individually fed to receive 0.5% (MAINT) or 1.25% [High gain (HG)] of their BW daily for 67 days. At the end of the feeding period, semen was collected through electroejaculation and frozen. Antral follicles were aspirated from ovaries obtained from a slaughterhouse and utilized for IVP in 4 independent replicates (n = 2,227 total oocytes). Cleavage rates were evaluated 48 h after fertilization and blastocyst development rates were evaluated after 7 days of embryo culture. The proposed treatments successfully induced differences in BW gain (P &lt; 0.01; 2.28 vs -0.04 kg/d) and carcass composition (Rump fat: 1.63 vs. 0.41 cm, P = 0.08; Rib fat: 1.06 vs. 0.41 cm, P = 0.02; intramuscular fat: 3.5 vs. 3.0%, P = 0.36; for HG vs. MAINT sires, respectively). There was a significant decrease in cleavage rates (69.9 ± 2.5 vs. 65.0 ± 2.7; P &lt; 0.04), blastocyst rate as a percentage of oocytes (16.7 ± 2.9 vs. 11.5 ± 2.1; P &lt; 0.01), and blastocyst rates as a percentage of cleaved structures (24.1 ± 3.8 vs. 11.5 ± 2.1; P &lt; 0.01) for HG compared with MAINT sires. In conclusion, sires fed diets that induce highly anabolic conditions had impaired blastocyst development compared to sires fed a maintenance diet.

242 USE OF BRAIN-DERIVED NEUROTROPHIC FACTOR IN IN VITRO PREMATURATION OF BOVINE OOCYTES SUBJECTED TO PARTHENOGENETIC ACTIVATION

2008 ◽  
Vol 20 (1) ◽  
pp. 200
Author(s):  
T. H. C. De Bem ◽  
R. Rochetti ◽  
P. R. L. Pires ◽  
F. F. Bressan ◽  
P. R. Adona ◽  
...  
Keyword(s):  
In Vitro Culture ◽  
Embryo Development ◽  
Polar Body ◽  
Hatching Rate ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Parthenogenetic Activation ◽  
Bovine Oocytes ◽  
Hatching Rates

Prematuration provides an additional time for oocyte capacitation and maturation in an attempt to improve in vitro embryo production (IVP) rates and allows media supplementation during this period for IVP. The aim of this study was to use brain-derived neurotropic factor (BDNF) in prematuration to improve maturation of bovine oocytes subjected to parthenogenetic activation and cultured with different media. Oocytes were subjected to prematuration in TCM-199 medium supplemented with 10 µm butyrolactone I, 2.0 mm pyruvate, and 10 µg mL–1 gentamicin for 24 h in the absence of BDNF (control) or in the presence of 10 ng mL–1 BDNF (BD). Oocytes were then in vitro-matured (IVM) in TCM-199 medium supplemented with 10% FCS, 0.5 µg mL–1 FSH, 5.0 µg mL–1 LH, 2.0 mm pyruvate, and 10 µg mL–1 gentamicin at 38.5�C under 5% CO2 in air. After 19 h oocytes were denuded using hyaluronidase and vortexing for 3 min for the 1st polar body (1PB) selection. Those which extruded the 1PB were maintained in IVM until 26 h, when parthenogenetic activation was performed (5 min in 5 µm ionomycin, followed by 3 h in 2 mm 6-DMAP). Activated oocytes were then transferred to in vitro culture (IVC) for embryo development evaluation. Embryos from both groups were cultured in SOF medium with 2.5% FCS, 0.05 g mL–1 BSA, 0.2 mm pyruvate, and 10 mg mL–1 gentamicin. Cleavage rates on the second day of in vitro culture (D2), embryo production at Days 7 and 8 (D7 and D8), and hatching rate at Day 8 were evaluated. Data regarding 1PB extrusion, cleavage, blastocyst development on D7 and D8, and blastocyst D8 hatching rates of three replicates were analyzed by chi-square test at 5% significance using the BIOESTATS 4.0 software. Control and BD, respectively, did not show differences (P > 0.05) regarding 1PB extrusion (n = 164, 63.81%, and n = 175, 66.79%) or cleavage (n = 117, 71.34%, and n = 138, 78.86%). However, for control and BD, respectively, blastocyst development on D7 (n = 63, 38.41%, and n = 89, 50.86%), D8 (n = 63, 38.41%, and n = 91, 52.00%), and hatching on D8 (n = 22, 34.92%, and n = 39, 43.82%) were all significantly higher for BD when compared with control (P < 0.05). In conclusion, BDNF during prematuration improved in vitro embryo development by increasing blastocyst and hatching rates of parthenogenetic embryos.

scholarly journals 287IN VITRO PRODUCTION OF HOLSTEIN EMBRYOS USING SEX SORTED SEMEN

2004 ◽  
Vol 16 (2) ◽  
pp. 263
Author(s):  
R.D. Wilson ◽  
K.A. Weigel ◽  
P.M. Fricke ◽  
M.L. Leibfried-Rutledge ◽  
D.L. Matthews ◽  
...  
Keyword(s):  
Low Cost ◽  
Laboratory Data ◽  
Blastocyst Stage ◽  
Embryo Production ◽  
Cull Cows ◽  
Sexed Semen ◽  
Dairy Cattle Breeding ◽  
In Vitro Embryo Production ◽  
Freeze Date

Our objective was to explore the synergy between sexed semen and in vitro embryo production and assess benefits of these technologies on commercial farms. Ovaries were collected from high genetic merit Holstein cull cows via colpotomy or at the time of slaughter. Oocytes were aspirated from the ovaries, fertilized 20–24h later, and matured to the morula or blastocyst stage. Embryos were transferred into recipient Holstein cows and heifers on the same farms. Seven Wisconsin herds participated, and 365 embryos were produced from 104 donor cows. Only 272 of these embryos were transferred due to limited availability of recipients. Sexed semen from three Holstein sires was used. On average, 3.5±0.37 transferable embryos were produced per donor, including 1.4±0.18 grade 1 embryos and 1.5±0.20 grade 2 embryos. Individual farms averaged from 1.6 to 5.8 transferable embryos per donor. Laboratory data also revealed interesting results. On average 43.7±4.0 oocytes were collected per donor, and the number of usable oocytes (33.9±3.4), and percent embryos cleaved (52.1±1.9), were significant predicators of the number of blastocysts developed. We divided the usable oocytes and embryos cleaved per donor into quartiles. The fourth quartile for embryos cleaved was significantly greater (P&lt;0.05) than the lower three quartiles, and the usable oocyte quartiles all significantly differed from each other. Semen freeze date was also a significant predicator of the number of blastocysts developed, suggesting significant variation in the quality of sorted semen per ejaculate. To preliminarily test the effect of sorting on the percentage of embryos developing to blastocyst stage, oocytes were recovered from ovaries collected at a slaughterhouse and fertilized using non-sorted semen or sex-sorted semen from the same sires. Oocytes (n=3312) fertilized using non-sorted semen tended (P=0.06) to produce more embryos developing to blastocyst stage than oocytes (n=1577) fertilized using sex-sorted semen (20.1±2.9% v. 12.2±2.3%, respectively). Preliminary pregnancy results show strong farm and sire effects. Overall conception rate was 36% for heifer recipients and 18milking cow recipients. These results suggest that low cost in vitro embryo production may have promise as an early system for utilizing sexed semen in dairy cattle breeding programs.

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

scholarly journals Oocyte Source and Hormonal Stimulation forIn VitroFertilization Using Sexed Spermatozoa in Cattle

2011 ◽  
Vol 2011 ◽  
pp. 1-8 ◽  
Author(s):  
Giorgio A. Presicce ◽  
Jie Xu ◽  
Guochun Gong ◽  
Juan F. Moreno ◽  
Sanjeev Chaubal ◽  
...  
Keyword(s):  
Hormone Treatment ◽  
Intramuscular Administration ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Hormonal Stimulation ◽  
In Vitro Embryo Production ◽  
Donor Oocytes

The aim of this study was to investigate the efficiency of in vitro embryo production in cattle utilizing sexed sperm from two bulls and oocytes recovered by OPU. Twenty donor animals were employed in eight OPU replicates: the first four OPU trials were conducted on animals without hormone treatment, and the last four were run on the same animals, following FSH subcutaneous and intramuscular administration. A higher rate of blastocyst development was recorded in stimulated, as compared to nonstimulated animals, (25.2% versus 12.8%, ). Ocytes derived from slaughterhouse (SH) ovaries were also fertilized with sperm from the same bulls. Overall, non-sexed sperm used with oocytes derived from SH ovaries was significantly more efficient for blastocyst development than was sexed sperm with these same SH derived oocytes and sexed sperm with stimulated donor oocytes (39.8% versus 25.0% and 25.2%, ). In conclusion, the use of sexed sperm with OPU-derived oocytes resulted in a significantly higher blastocyst development when donors were hormonally stimulated; furthermore, the level of efficiency achieved was comparable to that attained when the same sexed sperm was tested on oocytes derived from SH ovaries.

scholarly journals Use of strontium in the activation of bovine oocytes reconstructed by somatic cell nuclear transfer

Zygote ◽  
2005 ◽  
Vol 13 (4) ◽  
pp. 295-302 ◽  
Author(s):  
Walt Yamazaki ◽  
Christina Ramires Ferreira ◽  
Simone Cristina Méo ◽  
Cláudia Lima Verde Leal ◽  
Flávio Vieira Meirelles ◽  
...  
Keyword(s):  
Somatic Cell ◽  
Nuclear Transfer ◽  
Standard Treatment ◽  
Oocyte Activation ◽  
Blastocyst Development ◽  
Reconstructed Embryos ◽  
Developmental Capacity ◽  
Bovine Oocytes

As an important step in the nuclear transfer (NT) procedure, we evaluated the effect of three different treatments for oocyte activation on the in vitro and in vivo developmental capacity of bovine reconstructed embryos: (1) strontium, which has been successfully used in mice but not yet tested in cattle; (2) ionomycin and 6-dimethylaminopurine (6-DMAP), a standard treatment used in cattle; (3) ionomycin and strontium, in place of 6-DMAP. As regards NT blastocyst development, no difference was observed when strontium (20.1%) or ionomycin/6-DMAP (14.4%) were used. However, when 6-DMAP was substituted by strontium (3), the blastocyst rate (34.8%) was superior to that in the other activation groups (p <0.05). Results of in vivo development showed the possibility of pregnancies when NT embryos activated in strontium were transferred to recipient cows (16.6%). A live female calf was obtained when ionomycin/strontium were used, but it died 30 days after birth. Our findings show that strontium can be used as an activation agent in bovine cloning procedures and that activation with a combination of strontium and ionomycin increased the in vitro developmental capacity of reconstructed embryos. This is the first report of a calf produced by adult somatic cell NT in Latin America.

365 QUALITY ASSESSMENT OF BOVINE CRYOPRESERVED SPERM AFTER SEXING BY FLOW CYTOMETRY AND ITS USE FOR IN VITRO EMBRYO PRODUCTION

2010 ◽  
Vol 22 (1) ◽  
pp. 339
Author(s):  
J. O. Carvalho ◽  
R. Sartori ◽  
G. M. Machado ◽  
G. B. Mourão ◽  
M. A. N. Dode
Keyword(s):  
Flow Cytometry ◽  
Sperm Motility ◽  
Membrane Integrity ◽  
Fertilization Rate ◽  
Computer Assisted ◽  
Blastocyst Development ◽  
Embryo Production ◽  
Acridine Orange Staining ◽  
In Vitro Embryo Production

Several studies using sex-sorted sperm by flow cytometry have shown that its fertility is reduced. Therefore, this study evaluated structural and functional characteristics of sperm sexed by flow cytometry. In addition, in vitro embryo production (IVP) and development was assessed when frozen-thawed unsorted and sex-sorted sperm from 4 Nellore bulls. Each ejaculate was separated into three fractions: non-sexed (NS), sexed for X-sperm (SX), and sexed for Y-sperm (SY). After thawing, each sample was analyzed for sperm motility by computer-assisted semen analysis (CASA, Berkeley, CA), sperm head agglutination, sperm morphology, membrane integrity by propidium iodide (PI) and 6-carboxy-fluorescein diacetate (CFDA) staining, acrosome integrity by peanut agglutinin (PNA), capacitation by chlortetracycline (CTC), and chromatin integrity by acridine orange staining. Then, the samples were placed in 45 : 90% (NS90) or 45 : 60% (NS60, SX, and SY) Percoll™ gradients. After Percoll™ centrifugation, sperm pellets were analyzed or used for IVP. All analyses were replicated independently three times. For IVP, 2,271 in vitro matured oocytes were used. To assess fertilization rate, presumptive zygotes were fixed and stained with lacmoid at 18 h post-insemination (hpi). Cleavage was evaluated at Day 2 (48 hpi) and blastocyst development at Days 6, 7, 8, and 9 of culture. Data were analyzed using generalized linear models. No differences (P > 0.05) were observed between SX and SY groups for e sperm variables evaluated either before or after Percoll™. However, non-sexed sperm had higher sperm motility, greater percentage of sperm with intact membranes, and greater percentage of live sperm with intact acrosomes than sexed sperm (P < 0.05). An effect of Percoll™ was observed in the non-sexed samples, with those submitted to 45 : 90% gradient having higher motility, greater percentage of cells with intact membrane, and lower recovery rate than those submitted to a 45 : 60% gradient. No differences among groups were observed for fertilization rate, being 74.0 ± 5.7, 63.2 ± 5.1, 67.2 ± 5.7, and 55.4 ± 5.9% for NS90, NS60, SX, and SY, respectively. Group NS90 showed a greater cleavage rate than did the SY group, while groups NS60 and SX had similar rates to the others. Blastocyst development rates on Day 6 to Day 9 were greater for group NS90. For example, on Day 8 the blastocyst rate was 34.9 ± 3.6, 22.2 ± 3.2, 18.1 ± 3.3, and 14.8 ± 2.9% forNS90, NS60, SX, and SY groups, respectively. All groups showed similar embryonic developmental stages on Day 6 to Day 9. Although sex-sorting affected sperm characteristics, it did not cause a decrease with in vitro fertility. However, differences in blastocyst rates between groups NS60 and NS90 indicated that the sperm selection protocol affected embryo production. Financial support: Embrapa Genetic Resources and Biotechnology.

207 DEVELOPMENT OF A ROUTINE IN VITRO EMBRYO PRODUCTION SYSTEM USING SINGLY IN VITRO-MATURED, FERTILIZED, AND CULTURED BOVINE OOCYTES

2009 ◽  
Vol 21 (1) ◽  
pp. 201
Author(s):  
I. G. F. Goovaerts ◽  
J. L. M. R. Leroy ◽  
J. B. P. De Clercq ◽  
S. Andries ◽  
P. E. J. Bols
Keyword(s):  
Production System ◽  
Cumulus Cell ◽  
Developmental Competence ◽  
Control Individual ◽  
Embryo Production ◽  
Group Culture ◽  
In Vitro Embryo Production ◽  
Bovine Oocytes ◽  
Hatching Rates

An in vitro embryo production system (IVP), in which a single oocyte can be tracked from the moment of retrieval up to the blastocyst stage, would be a valuable tool for studies linking developmental competence and embryo metabolism to oocyte quality and follicular environment. Unfortunately, to date, data on individual IVP are inconsistent, and in most cases show unsatisfactory blastocyst rates. Earlier studies revealed that individual culture on a cumulus cell (CC) monolayer resulted in comparable numbers of good-quality embryos as obtained after regular group culture (Goovaerts et al. 2008 Reprod. Dom. Anim. 43 (Suppl. 3), 190). Because, in the latter study, single culture was performed after group maturation and fertilization, the aim of this study was to develop and test an individual IVP system using bovine oocytes or zygotes obtained after single maturation and single fertilization. Therefore, 532 grade I COC from slaughterhouse ovaries (3 replicates) were randomly assigned to 1 of 2 treatments: a complete individual IVP protocol, or a routine group IVP as a control. Individual maturation (TCM-199 + 20% serum) and fertilization were performed in 20-μL droplets under oil in 24-well plates. Subsequently, each zygote was cultured in 20 μL of medium (SOF + 5% serum, 90% N2, 5% CO2, 5% O2) on a 6-day-old monolayer of matured CC (5% CO2 in air). Group maturation and fertilization were carried out per 100 COC in 500 μL, whereas group culture was performed per 25 zygotes in 50-μL droplets under oil. Cleavage, blastocyst, and hatching rates were assessed 2, 8, and 10 days postfertilization, respectively. Possible effects of individual and group culture were evaluated with binary logistic regression (SPSS 15.0). No interactions between replicate and treatment could be found (P > 0.05). Cleavage and blastocyst rates were significantly lower after individual IVP, compared with group IVP, whereas the blastocyst rates on cleaved zygotes and the hatching rates did not differ significantly (Table 1). In conclusion, acceptable blastocyst rates (25.1%) could be obtained after individual IVP. The lower blastocyst rates were associated with the lower cleavage rates, and no effect of the individual embryo culture system on embryo development could be found. Table 1.Cleavage, blastocyst, and hatching rates after individual and group in vitro embryo production (IVP)

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.

Sign in / Sign up

Export Citation Format

Share Document