Preliminary results from the use of FACS-sorted bull sperm in an in vitro embryo production system

1994 ◽  
Vol 41 (1) ◽  
pp. 210
Author(s):  
M.P. Gurnsey ◽  
L.J. Hagemann ◽  
Dingyi Xu ◽  
R.A.S. Welch
Keyword(s):  
Production System ◽  
Embryo Production ◽  
Preliminary Results ◽  
In Vitro Embryo Production ◽  
Bull Sperm

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

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

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

Virus screening of a bovine in vitro embryo production system

1993 ◽  
Vol 132 (26) ◽  
pp. 660-660 ◽  
Author(s):  
B. Avery ◽  
T. Greve ◽  
L. Ronsholt ◽  
A. Botner
Keyword(s):  
Production System ◽  
Embryo Production ◽  
In Vitro Embryo Production ◽  
Virus Screening

scholarly journals 245 OOCYTE RECOVERY BY OVUM PICK UP AND EMBRYO PRODUCTION IN MURRAH AND NILI-RAVI BUFFALOES (BUBALUS BUBALIS) IMPORTED IN CHINA

2005 ◽  
Vol 17 (2) ◽  
pp. 273 ◽  
Author(s):  
Y. Huang ◽  
X. Zhang ◽  
B. Gasparrini ◽  
G.A. Presicce
Keyword(s):  
Essential Amino Acids ◽  
Reproductive Technologies ◽  
Meat Production ◽  
Embryo Production ◽  
Vitro Fertilization ◽  
Preliminary Results ◽  
Swamp Buffaloes ◽  
In Vitro Embryo Production ◽  
Humidified Air

In this preliminary study, in vitro embryo production and cryopreservation in two river type buffaloes (Murrah and Nili-Ravi) imported into China have been carried out. The objective of the study was enhancement of the genetic merit and productive performances of imported river buffaloes in conjunction with the utilization of local swamp buffaloes. In order to improve milk and meat production in China local swamp buffaloes (2n = 48), which are the predominant subspecies, have been crossbred with imported river buffaloes (Murrah and Nili-Ravi: 2n = 50). At present, several hundred thousand crossbred heads have been produced, and although both males and females can reproduce with 2n = 49 crossbred buffaloes, their reproductive performances are significantly reduced when compared to 2n = 50 buffaloes. As an alternative approach, a program of embryo production in river buffaloes and transfer into both river and swamp buffaloes has been implemented at the Guangxi Buffalo Research Institute, in Nanning, P.R. China. Some preliminary results are presented: from a start-up experiment, a total of 46 river buffaloes were subjected to 2 to 3 ovum pickup sessions at 4-day intervals. A total of 750 antral follicles were punctured and 495 (66%) cumulus-oocyte complexes (COCs) were retrieved. Only COCs characterized by at least one layer of granulosa cells (n = 451; 91.1%) were considered for in vitro maturation (IVM). COCs were matured in TCM 199 + 10% FCS, 0.5 μg/mL FSH, 5 μg/mL LH, and 1 μg/mL estradiol in the presence of cysteamine (50 μM) at 39°C under 5% CO2 in humidified air for 24 h. Of the initial 451 COCs matured, only 277 could be considered for in vitro fertilization (IVF). IVF was performed at 39°C under CO2 in humidified air in TALP medium supplemented with 0.2 mM penicillamine, 0.1 mM hypotaurine, and 0.01 mM heparin. Frozen/thawed sperm from a tested bull was treated by swim-up procedure and used at a final concentration of 20 million/mL. Following 20 to 22 h of co-incubation, presumptive zygotes were cultured in SOF medium, supplemented with essential and non-essential amino acids and 8 mg/mL BSA, in a gas atmosphere of 5% CO2, 7% O2, and 88% N2. A total of 41 (14.8%) blastocysts were produced, of which 33 were vitrified and 8 transferred immediately into available swamp and river buffalo recipients. Two calves were born (25%) from the transfer of fresh embryos into one river and one swamp buffalo. In vitro embryo production in the buffalo species is still characterized by a high degree of variable results. However, these preliminary results reinforce the need to implement newly developed reproductive technologies not only for speeding up genetic gain of already productive species, but also for the utilization of local breeds characterized by reduced productive performance.

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)

Fingerprints on oocyte and embryo lipid profiles caused by in vitro embryo production system and fatty acid diet supplementation in bovine

2014 ◽  
Author(s):  
Serrano Andres Felipe Gonzalez ◽  
Christina R Ferreira ◽  
Valentina Pirro ◽  
Andrea Lucas-Hahn ◽  
Julia Heinzmann ◽  
...  
Keyword(s):  
Fatty Acid ◽  
Production System ◽  
Lipid Profiles ◽  
Embryo Production ◽  
Acid Diet ◽  
In Vitro Embryo Production ◽  
Diet Supplementation

297 ADDITION OF GLUTATHIONE TO THAWING MEDIUM FOR BULL SPERMATOZOA IMPROVES THE IN VITRO EMBRYO PRODUCTION

2007 ◽  
Vol 19 (1) ◽  
pp. 264
Author(s):  
L. Grullón ◽  
J. C. Gardó ◽  
F. García-Vázquez ◽  
J. Gadea
Keyword(s):  
Oxidative Stress ◽  
Defense Mechanism ◽  
Developmental Stages ◽  
Ros Generation ◽  
Cleavage Rate ◽  
Embryo Production ◽  
Short Period ◽  
In Vitro Embryo Production ◽  
Bull Sperm

The cold shock of spermatozoa is associated with oxidative stress induced by reactive oxygen species (ROS) generation (Chatterjee et al. 2001 Mol. Reprod. Dev. 60, 498–506). Glutathione (l-�-glutamyl-l-cysteinylglycine; GSH) plays an important role as an intracellular defense mechanism against oxidative stress. The process of freezing is associated with a significant reduction in GSH content in novine spermatozoa (Bilodeau et al. 2000 Mol. Reprod. Dev. 55, 282–288). The main objective of this study was to evaluate the effect of GSH supplementation of the thawing extender on bull sperm used for in vitro embryo production. To examine the effect of GSH supplementation during the thawing process, spermatozoa from 6 different bulls were incubated without addition of GSH (control), and with addition of 1 mM or 5 mM GSH to the sperm-TALP medium, and maintained 30 min at 37�C in these media before assay. IVM and IVF were performed as described Coy et al. (2005 Reproduction 129, 19–26, 747–755). Oocytes were fertilized with frozen–thawed semen (106 total sperm/mL). After 18 h co-incubation, they were cultured in KSOM medium for 7 days. Embryos were examined for developmental stages at Day 2 (2–4 cells), Day 3 (8–16 cells), and Day 7 (morula–blastocysts). Finally, the total number of nuclei of blastocysts was determined by staining with Hoechst 33342 (10 mg mL-1; 20 min). The percentage of cleavage in the different embryo stages was higher in the 5 mM GSH group [n = 233, Day 2 (64.63%), Day 3 (56.92%), and Day 7 (31.28%)] than in control [n = 229, Day 2 (47.64%), Day 3 (42.16%), and Day 7 (22.32%)], and the 1 mM group maintained an intermediate position with [n = 246, Day 2 (54.88%), Day 3 (49.56%), and Day 7 (28.98%)]. The bull sperm affected significantly the cleavage parameters studied. The number of nuclei within the blastocysts ranged from 39.93 to 47.71, and no differences were observed between experimental groups. It is known that addition of GSH produces a reduction in ROS generation (Gadea et al. 2005 J. Androl. 26, 749–756). The results showed that in the treatment group there is a reduction in ROS generation in spermatozoa and an improvement in the cleavage rate and blastocyst formation when GSH was used. This study demonstrates that only a short period of incubation of the spermatozoa in an antioxidant could improve the spermatozoa functionality of producing more and better embryos. This work was supported by AGL2003-03144, BIOCARM 10BIO2005/01-6463 and Seneca 03018/PI/05.

The contribution of the male to ovine embryogenesis in an in vitro embryo production system

2003 ◽  
Vol 75 (1-2) ◽  
pp. 9-26 ◽  
Author(s):  
Lee H.A Morris ◽  
Adrienne E Randall ◽  
W.A King ◽  
W.H Johnson ◽  
B.C Buckrell
Keyword(s):  
Production System ◽  
Embryo Production ◽  
In Vitro Embryo Production

scholarly journals Laparoscopic Ovum Pick-Up Followed by In Vitro Embryo Production and Transfer in Assisted Breeding Programs for Ruminants

Animals ◽  
2021 ◽  
Vol 11 (1) ◽  
pp. 216
Author(s):  
Hernan Baldassarre
Keyword(s):  
Pregnancy Rate ◽  
Commercial Application ◽  
Embryo Production ◽  
Breeding Programs ◽  
Sheep And Goats ◽  
Embryo Recovery ◽  
Marker Selection ◽  
Holstein Calves ◽  
In Vitro Embryo Production

The potential of laparoscopic ovum pick-up (LOPU) followed by in vitro embryo production (IVEP) as a tool for accelerated genetic programs in ruminants is reviewed in this article. In sheep and goats, the LOPU-IVEP platform offers the possibility of producing more offspring from elite females, as the procedure is minimally invasive and can be repeated more times and more frequently in the same animals compared with conventional surgical embryo recovery. On average, ~10 and ~14 viable oocytes are recovered by LOPU from sheep and goats, respectively, which results in 3–5 transferable embryos and >50% pregnancy rate after transfer. LOPU-IVEP has also been applied to prepubertal ruminants of 2–6 months of age, including bovine and buffalo calves. In dairy cattle, the technology has gained momentum in the past few years stemming from the development of genetic marker selection that has allowed predicting the production phenotype of dairy females from shortly after birth. In Holstein calves, we obtained an average of ~22 viable oocytes and ~20% transferable blastocyst rate, followed by >50% pregnancy rate after transfer, declaring the platform ready for commercial application. The present and future of this technology are discussed with a focus on improvements and research needed.

Sign in / Sign up

Export Citation Format

Share Document