363 ENHANCEMENT OF COMMERCIAL BOVINE IN VITRO EMBRYO PRODUCTION BY SEXING FROZEN SEMEN PRIOR TO INSEMINATION

2010 ◽  
Vol 22 (1) ◽  
pp. 338
Author(s):  
D. B. Ardais ◽  
L. T. S. Yamazaki ◽  
L. P. Landim Junior ◽  
E. C. D. Benzi ◽  
D. P. Corneglian ◽  
...  
Keyword(s):  
Production Rate ◽  
Culture Media ◽  
Bovine Embryos ◽  
Embryo Production ◽  
Efficient System ◽  
Frozen Semen ◽  
In Vitro Embryo Production ◽  
Female Cattle ◽  
Better Than

The high commercial demand on production of genetically valuable female cattle was emphasized after use of sex-sorted semen in commercial in vitro embryo production (IVEP) programs and a reduction of interest in the use of embryo biopsy followed by PCR. Brazil is a leader in IVEP of bovine embryos, and frozen-thawed, sex-sorted samples become a new option for embryo sexing. There are only a few reports in the literature using this new technique for IVEP. Most reports have involved the use of frozen-thawed then sorted for use with AI in Europe. In the present study, frozen semen from three Nelore bulls was used and the blastocyst production rate was evaluated at Day 7 of IVP and compared to previous data from the same bulls and a batch of non-sorted semen. Cumulus oocyte complexes obtained from high genetic merit donors by ovum pickup were matured (TCM-199, supplemented with FCS, LH, FSH, estradiol, pyruvate, and antibiotics) for 24 h and fertilized (Fert-TALP supplemented with BSA, phenylalanine, and heparin) for 18-22 h (Day 0) in vitro. Frozen semen straws were thawed at 35°C for 30 s in a water bath and then selected by centrifugation at 800 g on discontinuous Percoll™ gradients (45 :90%). Samples of frozen-thawed sex-sorted semen were processed from straws from bulls that in some cases may already be dead. Frequently, 3 to 5 straws were thawed, flow-sorted, and then centrifuged to provide sufficient sperm to fertilize 100 oocytes. On Day 1, presumptive zygotes were transferred to culture media (SOFaa supplemented with BSA and FCS) and on Day 7 blastocyst production rate was evaluated. The present results were commercially satisfactory because the frozen-thawed sex-sorted semen from all three bulls performed comparably and, in one case, better than non-sorted semen from the same bulls. Therefore, its use is extremely attractive in commercial bovine IVEP systems and is a more efficient system than embryo sexing. Table 1.Embryo production rates at 7 days post insemination

178 IMPROVED DEVELOPMENTAL COMPETENCE OF BOVINE EMBRYOS BY PGI2 ANALOG TREATMENT

2006 ◽  
Vol 18 (2) ◽  
pp. 197 ◽  
Author(s):  
B. S. Song ◽  
J. S. Kim ◽  
D. B. Koo ◽  
J. S. Park ◽  
K. K. Lee ◽  
...  
Keyword(s):  
Inner Cell Mass ◽  
Culture Media ◽  
Cell Mass ◽  
Developmental Rate ◽  
Treated Group ◽  
Developmental Competence ◽  
Bovine Embryos ◽  
Frozen Semen ◽  
Oviductal Fluid

The microenvironment of the follopian tube, in which the oviductal fluid contains a variety of cytokines and growth factors, affects pre-implantation development of fertilized embryos in mammals. Prostaglandin I2 (PGI2, prostacyclin) exists in oviductal fluid and is synthesized from arachidonic acid by prostacyclin synthetase. PGI2 also enhances the implantation rate of mouse embryos. In this study, the effect of PGI2 analog on the development of bovine embryos was examined. Bovine cumulus oocytes complexes (COCs) were matured in TCM-199 medium supplemented with 10 IU/mL pregnant mare serum gonadotropin (PMSG), 10 IU/mL hCG, and 10 ng/mL epidermal growth factor (EGF) at 39�C, 5% CO2 in air for 20-22 h. Following in vitro maturation, COCs were fertilized in Fert-TALP medium containing 0.6% BSA using frozen semen. Also, oocytes matured in vitro were enucleated, individually reconstructed with bESF cells, fused, and then activated by treatment with 5 �M ionomycin for 5 min and 2 mM 6-DMAP for 4 h. In vitro-fertilized (IVF) and nuclear-transferred (NT) eggs were cultured in 50 ��L drops of CR1-aa medium supplemented with 0.3% BSA in the absence or presence of 1 �M PGI2 analog at 39�C, 5% CO2 in air, respectively. At 3 days of culture, cleaved embryos were further cultured in the same culture media supplemented with 10% FBS for 4 days. Allocations of blastocysts to inner cell mass (ICM) and trophoblast (TE) cells were investigated to assess embryo quality. All experiments were repeated more than three times. All data were analyzed by using the Duncan test of ANOVA by the Statistical Analysis System (SAS Institute, Inc., Cary, NC, USA) and numbers of nuclei in blastocysts were expressed as mean � SE. No difference was detected in the cleaved rate of the eggs between the treated- and nontreated groups. IVF zygotes treated with PGI2 analog represented a higher developmental rate (33%, 122/418) to the blastocyst stage than nontreated controls (24%, 107/456) (P < 0.05). Among IVF-derived blastocysts, interestingly, the proportion (46%, 84/181) of expanded blastocysts was significantly higher in the PGI2 analog-treated group compared with that in the nontreated group (28%, 46/164). The number of nuclei in (165 � 6.1, n = 15) in blastocysts in the PGI2 analog-treated group was higher than that (146.12 � 5.7, n = 18) in the nontreated group (P < 0.05). No difference was detected in the ratio of ICM to total cells between PGI2 analog-treated (42.0 � 3.0%) and nontreated groups (41.9 � 2.9%). Like the IVF embryos, NT embryos in the PGI2 analog-treated group showed a higher in vitro developmental rate (33.6%, 43/128) than the nontreated embryos (24.2%, 32/132) (P < 0.05). Our results indicate that PGI2 analog improves the kinetics of embryo development in cattle.

238 KNOCKING DOWN OF THYROID HORMONE RECEPTORS INHIBITS DEVELOPMENT OF EARLY BOVINE EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 267
Author(s):  
N. Y. Rho ◽  
F. A. Ashkar ◽  
T. Revay ◽  
P. Madan ◽  
W. A. King
Keyword(s):  
Embryo Development ◽  
Reference Genes ◽  
Messenger Rna ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Blastocyst Formation ◽  
Embryo Production ◽  
Mrna Transcripts ◽  
In Vitro Embryo Production

Thyroid hormones (TH) play an important role in the physiology of vertebrates, ranging from the regulation of metabolic processes to cell proliferation, differentiation, and embryo development. We have previously shown a beneficial effect of supplementing TH in in vitro embryo production media. Recently, detection of TH receptors (TR) in oocytes and early stages of pre-implantation embryos indicated a possible regulatory role for TH in these stages (unpublished data). The objective of this study was to investigate the importance of TR expression in the pre-attachment bovine embryo in vitro. Bovine embryos, produced by standard in vitro embryo production procedures, were microinjected at the zygote stage with small interfering RNA (siRNA) specifically designed for knocking down either TR-α or TR-β. In addition, groups of zygotes were microinjected with scrambled siRNA (SI) or were not injected (NI), and these groups served as controls. Embryo developmental rates were assessed using light microscopy for blastocyst formation rates and expression of TR messenger RNA (mRNA) transcripts at the blastocyst stage was assessed by quantitative PCR across all groups. Expression of TR mRNA was normalized against glyceraldehyde 3-phosphate dehydrogenase, H2a, and 18S as reference genes. There was a significant decrease in blastocyst formation rates in both embryo groups injected with either TR-α (P < 0.002) and TR-β (P < 0.001) siRNA compared with the NI and SI groups. Moreover, the TR-β knockdown group exhibited a lower developmental rate than the TR-α knockdown group, which indicates a stronger inhibitory role for TR-β. Quantification of the level of TR mRNA expression in four groups normalized with three different reference genes shows a consistent significant reduction in the levels of TR-α (P < 0.05) and TR-β (P < 0.02) mRNA transcripts compared with the NI and SI groups. However, TR-β expression was inhibited more than was TR-α expression. In conclusion, the results indicate that knocking down either TR-α or TR-β restrains embryo development. This suggests that TH play a vital role in the regulation of embryo development through their receptors during bovine early embryogenesis. The specific role of each of these receptors and their mechanism of action in mediating development needs to be further elucidated. Funding was provided by CRC, NSERC, and the EmbryoGENE network.

72 Effect of incubation temperature and of CO2 concentration during early cleavage on equine in vitro embryo production

2019 ◽  
Vol 31 (1) ◽  
pp. 161
Author(s):  
J. Brom-de-Luna ◽  
R. Salgado ◽  
H. Canesin ◽  
M. Diaw ◽  
K. Hinrichs
Keyword(s):  
Room Temperature ◽  
Incubation Temperature ◽  
Culture Media ◽  
Co2 Concentration ◽  
Mixed Gas ◽  
Early Cleavage ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
Co2 Treatment

Intracytoplasmic sperm injection (ICSI) is currently being used for equine in vitro embryo production for both research and clinical purposes. However, some basic parameters for in vitro embryo production, such as the optimum incubator temperature and the optimum CO2 concentration/pH of medium for early embryo development, have not yet been established in the horse. The incubation temperature used by many laboratories for equine in vitro embryo production is 38.2°C, whereas the range of normal rectal temperature in the horse is 37.2 to 38.3°C. In Exp. 1, we evaluated maturation, cleavage, and blastocyst rates under 3 different culture temperatures. Cumulus-oocyte complexes were recovered from slaughterhouse-derived ovaries and shipped overnight at room temperature. Oocyte maturation was performed concurrently in separate incubators set to 37.2, 37.7, or 38.2°C. Mature oocytes were subjected to ICSI, then cultured in mixed gas (6% CO2, 5% O2, and remainder N2) at the same temperature at which they were matured. Embryo culture media used were a commercial human medium (global) for Days 0 to 5, then DMEM/F-12 from Day 5 to 10, both with 10% fetal bovine serum (FBS). In Exp. 2, we evaluated 3 different CO2 concentrations (6, 6.5, or 7% CO2) in mixed gas for the Day 0 to 5 culture in global+FBS, at 38.2°C. Cumulus-oocyte complexes were recovered from live mares by transvaginal aspiration and held overnight at room temperature; all other parameters remained the same as for Exp. 1. Data were analysed by Fisher’s exact test. In Exp. 1, a total of 280 oocytes were utilised; the outcomes for the 37.2, 37.7, and 38.2°C treatments were, respectively: maturation rates of 33, 38, and 42%; cleavage rates of 84, 86, and 88%; and blastocyst rates per injected oocyte of 35, 44, and 44%. There were no significant differences among the 3 temperature treatments for any parameter (P&gt;0.2). In Exp. 2, the pH of the global+FBS medium was 7.38, 7.35, and 7.3 for 6, 6.5, and 7% CO2, respectively. A total of 106 mature oocytes underwent ICSI; the outcomes for the 3 CO2 atmospheres, respectively, were cleavage rates of 68, 80, and 70% and blastocyst rates per injected oocyte of 42, 54, and 27%. The blastocyst rate for the 7% CO2 treatment was significantly lower than that for the 6.5% CO2 treatment (P&lt;0.05). These results indicate that equine in vitro embryo production is equally effective throughout the range of normal equine body temperature, and that equine blastocyst production is sensitive to small changes in CO2 atmosphere/pH of medium during early cleavage-stage development. Research was supported by the Clinical Equine ICSI Program, Texas A&amp;M University, the Link Equine Research Endowment, Texas A&amp;M University, and Fonds en Santé Équine (FSÉ), Faculté de Médecine Vétérinaire, and Université de Montréal.

292 EVALUATION OF THREE PROTEIN SOURCES FOR THE IN VITRO PRODUCTION OF BOVINE EMBRYOS

2010 ◽  
Vol 22 (1) ◽  
pp. 302
Author(s):  
E. A. Ordoñez-Leon ◽  
G. Cancino ◽  
J. Hernandez-Ceron ◽  
J. A. Medrano ◽  
Y. C. Ducolomb ◽  
...  
Keyword(s):  
Embryo Development ◽  
Culture Media ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
In Vitro Production ◽  
Embryo Production ◽  
Serum Free ◽  
Serum Replacement ◽  
Knockout Serum Replacement

Bovine embryo development in vitro can be affected by many factors, including protein source, which can cause embryo development failure. The use of in vitro culture media supplemented with serum-free compounds could allow a better understanding of embryo requirements during the preimplantation stages by eliminating a highly variable and undefined compound such as serum. The objective of this study was to evaluate the effect of 3 different protein supplements used during IVM, IVF, and IVC on embryo production. Ovaries were collected from slaughtered cows and then aspirated to obtain oocytes for in vitro embryo production procedures. A total of 2056 oocytes were used, from which 685 were processed with maturation medium supplemented with 10% serum replacement (SR) (Gibco Knockout Serum Replacement, Invitrogen, Carlsbad, CA, USA), a defined serum-free formulation (TCM-199 + SR), fertilization medium with SR (TALP + SR), and culture medium with SR (SOF + SR). These were compared with 675 and 696 oocytes processed with the same IVM, IVF, and IVC media, but supplemented with 10% FCS or 10% heat-inactivated estrous cow serum (ECS), respectively. Data obtained from the variables studied were processed by analysis of variance and means were compared by Tukey’s test. The percentages of embryos produced with FCS (52.4%) and ECS (52.7%) were significantly higher compared with the percentage obtained with SR (41.5%) (P < 0.05). The percentages of morulae were similar in the groups supplemented with FCS (36.5%) and SR (36.7%), but significantly higher than the percentage in the ECS group (26.9%) (P < 0.05). For blastocysts, the percentages of embryos developed with FCS (35.2%) and ECS (35.6%) were significantly higher than that obtained with SR (29.2%) (P < 0.05). When evaluating expanded blastocysts, the percentage obtained in the FCS (45.9%) group was significantly higher than that in the ECS group (33.2%), and this was significantly higher than that obtained in SR (21%), with all these differences being significant (P < 0.05). It is concluded that it is possible to produce bovine embryos in vitro using FCS, ECS, or SR as supplements in IVM, IVF, and IVC media. Significant differences were found in different embryo stages, with the highest proportion of embryos developing with the addition of FCS, whereas supplementation with SR only improved the production of morulae. We thank Consejo Nacional de Ciencia y Tecnologia (CONACYT-Mexico) for the graduate student’s scholarship.

130 In vitro embryo production using frozen semen from cloned and non-cloned Bos indicus bulls

2019 ◽  
Vol 31 (1) ◽  
pp. 190
Author(s):  
S. Romo ◽  
O. Sebastián ◽  
F. Guerrero ◽  
R. Romero ◽  
F. Muñoz ◽  
...  
Keyword(s):  
Bos Indicus ◽  
The State ◽  
Assisted Reproductive Techniques ◽  
Embryo Production ◽  
Humid Atmosphere ◽  
Frozen Semen ◽  
Significant Difference ◽  
Reproductive Techniques ◽  
In Vitro Embryo Production

Reproductive biotechnology has continued to evolve rapidly, allowing the development of techniques to increase reproductive efficiency and contribute to the genetic improvement of cattle. Some of these techniques include the in vitro maturation and IVF of oocytes, sperm sexing and cloning. These modern assisted reproductive techniques can help produce offspring of desired genetic characteristics and of a pre-determined sex. However, studies of the bull’s contribution to in vitro reproductive performance are scarce in the Brahman breed. The aim of this study was to compare oocyte maturation and embryo production in vitro using frozen semen from 5 Brahman bulls (Bos indicus), cloned (n=1) and non-cloned (n=4), with characteristics and genetics of high commercial value. The age of the bulls at the time of semen collection and cryopreservation ranged from 2 to 7 years. The oocytes were obtained on 2 different dates (45 days between collections) using pooled oocytes collected by ovum pickup at random stages of the oestrous cycle, from a total of 15 Brahman donor cows. Oocytes were transported to a laboratory in the State of Chiapas, Mexico (Genemex Internacional). The oocytes were cultured in maturation medium for 24h. For IVF, conventional semen was used from one bull (B1) and his clone (B12), the grandson of B1 (B2), and 2 non-related bulls (B3 and B4). The gametes were co-incubated for 22h and afterward placed in medium for embryo development and cultured for 7 days in a humid atmosphere with 5% CO2 in air. Of the matured oocytes, 36/43 (84%), 16/32 (50%), 101/143 (70%), 46/67 (68%) and 53/65 (81%) were fertilized using semen from B1, B12, B2, B3 and B4, respectively. Of the fertilized oocytes, 15/30 (50%), 8/16 (50%), 45/101 (44%), 21/46 (45%) and 18/53 (34%) resulted in transferrable embryos, corresponding to semen from the same bulls, respectively. This would appear to be the first scientific report in Mexico about the use of semen from a cloned bull for in vitro embryo production. In IVF, similar results were observed between B1 and a non-related bull (B4). Similar results in transferrable embryos were observed between B1 and B12 but also similar to a related bull (B3) and a non-related bull (B4). A Fisher’s exact test of the IVF results comparing B1 and B12 found a significantly (P&lt;0.05) higher number of fertilized oocytes for B1. However, a significant difference was not found (P&gt;0.05) concerning the number of transferrable embryos produced by these two bulls. In conclusion, the Brahman bulls in this study differ in their contribution to IVF and embryo production. Further studies are required to determine the factors responsible for such effects, e.g. age differences or clone versus non-clone mosaicism. Results from this research contribute to the study and development of assisted reproductive techniques for increasing in vitro production efficiency in Zebu cattle. We thank the Rosales family, from El Herradero Ranch, in the State of Campeche, Mexico, for allowing the use of their cattle for this project.

Effect of donor stimulation, frozen semen and heparin treatment on the efficiency of in vitro embryo production in goats

2004 ◽  
Vol 62 (3-4) ◽  
pp. 576-586 ◽  
Author(s):  
Lucyna Kątska-Książkiewicz ◽  
Bożenna Ryńska ◽  
Barbara Gajda ◽  
Zdzisław Smorąg
Keyword(s):  
Embryo Production ◽  
Heparin Treatment ◽  
Frozen Semen ◽  
In Vitro Embryo Production

160 EFFECT OF BSA SOURCES AND TYPES ON IN VITRO-PRODUCED LIVESTOCK EMBRYO DEVELOPMENT

2006 ◽  
Vol 18 (2) ◽  
pp. 188
Author(s):  
J. R. Dobrinsky ◽  
A. M. Paprocki ◽  
V. L. Chrostowski ◽  
C. M. Penfield ◽  
K. R. Rozeboom ◽  
...  
Keyword(s):  
Reproductive Biology ◽  
Embryo Development ◽  
Bovine Serum ◽  
Culture Medium ◽  
Production Systems ◽  
Culture Media ◽  
Blastocyst Development ◽  
Embryo Production ◽  
In Vitro Embryo Production

Bovine serum albumin (BSA) is a macromolecule supplement used in embryo and cell culture media. Other chemicals have been used as macromolecule substitutes in embryo culture with variable effectiveness. There are BSA products available that are defined in their disease status, collection method, and manufacturing process. They are compliant for use in raw form or in culture medium in the USA and EU. It was the purpose of this study to compare the effectiveness of highly defined and internationally compliant BSA with typically used BSA on in vitro-produced pig and cow embryo development. Pig oocyte-cumulus complexes were matured in two stages for 44 h in vitro. Semen from one boar of known high fertility was used across the study to fertilize mature oocytes. After 6-h co-incubation with sperm (50 motile sperm/oocyte), presumptive zygotes were cultured for 120 h in modified NCSU-23 containing 4 mg/mL of one of the following heat-shocked BSA fractions: Sigma A-7906 (control), Minitube Reproductive Biology Grade Fraction-V (RBG-V1, RBG-V2, RBG-V3), or Minitube Reproductive Biology Grade Fatty Acid-Free (RBG-FAF1). Across all treatments, Day 5 morulae were removed from BSA culture and placed into modified NCSU-23 with 10% fetal bovine serum (FBS) (no BSA) and cultured for 48 h. After 168-h total culture, the following blastocyst development was observed: Sigma A-7906, 12.3% (40/324); RBG-V1, 21.1% (36/171); RBG-V2, 19.0% (30/158); RBG-V3, 16.8% (27/161); and RBG-FAF1, 13.4% (21/157). These data show that culture medium supplemented with Minitube Reproductive Biology Grade BSA meets or exceeds (P < 0.05; ANOVA-GLM of SAS; SAS Institute, Inc., Cary, NC, USA) blastocyst development potential when compared to culture medium supplemented with undefined standard BSA preparations, such as Sigma A-7906. In vitro-matured cow cumulus-oocyte complexes were fertilized and cultured in one of two CR-1aa-based media. Oocytes were fertilized in IVF medium containing 6 mg/mL Sigma A-8806 (FAF) or 6 mg/mL Minitube RBG-FAF1. After 24 h in IVF medium, presumptive zygotes from a specific BSA-supplemented medium were cultured for 144 h in CR-1aa supplemented with their respective BSA (8 mg/mL). Day 6 morulae were removed from BSA culture and placed into CR-1aa with 10% FBS (no BSA) and cultured for 48 h. After 192-h total culture, the following blastocyst development from oocytes matured was observed: Sigma A-8806, 21.4% (88/411); RBG-FAF1, 18.9% (70/370). These data show that culture medium supplemented with Minitube Reproductive Biology Grade BSA meets blastocyst development potential (P > 0.05) when compared to culture medium supplemented with undefined standard BSA preparations, such as Sigma A-8806. The inclusion of internationally compliant BSA meets or exceeds blastocyst development rates in comparison to standard BSA preparations in common in vitro embryo production systems for swine and cattle. Although manufacturing differences remain the prominent variant in BSA sources and types, continued monitoring and documentation of BSA preparations tested in livestock in vitro embryo production systems will ensure a safe global supply of BSA products for future culture media production.

scholarly journals 320EFFECT OF CYSTEAMINE DURING IN VITRO MATURATION ON FURTHER EMBRYONIC DEVELOPMENT AND POSTTHAW SURVIVAL OF IVP BOVINE EMBRYOS

2004 ◽  
Vol 16 (2) ◽  
pp. 279
Author(s):  
J.S. Merton ◽  
M. Gerritsen ◽  
D. Langenbarg ◽  
Z.L. Vermeulen ◽  
T. Otter ◽  
...  
Keyword(s):  
Embryonic Development ◽  
Production Rate ◽  
In Vitro Maturation ◽  
Survival Rates ◽  
Protective Role ◽  
Bovine Embryos ◽  
Embryo Production ◽  
In Vitro Embryonic Development ◽  
In Vitro Survival

The uptake of cysteamine by immature oocytes may facilitate the synthesis of glutathione (GSH) during in vitro maturation, as reported by Matos et al. (1995 Mol. Reprod. Dev. 42 432–436). GSH plays an important protective role in relation to reactive oxygen species generated by normal oxidative metabolism. This study investigated the effects of the presence of cysteamine during in vitro maturation on subsequent in vitro embryonic development and postthaw in vitro survival. Immature Cumulus-Oocyte-Complexes (COCs) were recovered from ovaries 6 to 8h after slaughter. COCs were matured in vitro for 22 to 24h in TCM199/FCS/LH/FSH supplemented either with or without cysteamine (0.1mM), Subsequently, matured oocytes were fertilized with frozen-thawed Percoll-separated semen and further cultured for seven days in SOFaaBSA. Morulae grade 1 (IETS) and blastocysts grades 1 and 2 (IETS) were frozen on Day 7 in 10% Glycerol using a conventional slow freezing procedure (Wagtendonk-de Leeuw et al. 1995 Cryobiology;; 32 157–167). In vitro survival was measured by rates of blastocyst formation and reexpansion at 24h and hatching/ed blastocysts at 72h in SOFaaBSA supplemented with 5% FCS. Results were analyzed by Chi-square analyses. The presence of cysteamine during in vitro maturation significantly affected the embryo production rate (19.4% and 24.0% for control and cysteamine at Day 7, respectively). The higher number of embryos at Day 7 was totally due to an increased number of blastocysts (Table 1); however, the distribution of embryos among the different quality grades was not affected. Addition of cysteamine did not affect the post thaw survival of the frozen/thawed embryos (85% v. 91% reexpansion and 33% v. 34% hatching/ed for control v. cysteamine, respectively). These results show that the presence of cysteamine during in vitro maturation, does affect further in vitro embryonic development, resulting in a higher embryo production rate. Embryo quality, expressed in morphological grades and postthaw survival rates, were not affected. A field trial will be conducted in order to confirm these results with ovum pick up-derived oocytes. Table 1 Effect of cysteamine during in vitro maturation on subsequent in vitro embryonic development of IVP bovine embryos (number of replicates: 5)

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