Effect of Percoll volume, duration and force of centrifugation, on in vitro production and sex ratio of bovine embryos

Machado, G. M.; Carvalho, J. F.; Filho, E. Siqueira; Caixeta, Ester Siqueira; Franco, M. M.; Rumpf, R.; Dode, M. A. N.

doi:10.1016/j.theriogenology.2009.01.002

Cited by 122 publications

(98 citation statements)

References 37 publications

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“…However, all four gradients preserved the expected 50:50 (male:female) ratio, consistent with previous studies using Percoll (Cesari et al 2006;Machado et al 2009). Although other studies have reported a difference in the sex ratio depending on the stage of development, since females develop slower than males (Rheingantz et al 2006), no such difference was proved in the present study.…”

Section: Vianna Et Al * Sperm Selection In Bovine Embryo Production 39supporting

confidence: 91%

Isolate^® and Optiprep^® minigradients as alternatives for sperm selection in bovine in vitro embryo production

et al. 2014

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. 2014. Isolate † and Optiprep † minigradients as alternatives for sperm selection in bovine in vitro embryo production. Can. J. Anim. Sci. 94: 35Á42. The objective of this study was to evaluate alternatives in small volumes to conventional gradient of Percoll † on semen quality, in vitro embryo production, sex ratio and embryo survival after vitrification. Thawed semen was randomly allocated to one of four density gradient selection methods: (1) conventional Percoll † (P), (2) MiniPercoll (MP), (3) MiniIsolate (MI), and (4) MiniOptiprep (MO). Sperm kinetics and quality were evaluated. Use of P, MP and MI gradients did not affect sperm motility (P!0.05). However, there was a decrease in total and progressive sperm motility in MO (70.8 and 51.3% vs. 87.3 and 69.5% for P; 87.3 and 73% for MP; 92.3 and 78.8% for MI; PB0.05). The MO had lower membrane integrity compared with P, MP and MI (39.7 vs. 70.5, 72.3, 63.8%, respectively, PB0.05). The percentage of blastocysts produced was higher in MI than in MP and MO (21.1 vs. 16.1 and 16.9%, PB0.05) and similar to P (18.4%; P!0.05). Sex ratio and embryo survival after vitrification were similar among groups (P!0.05). Semen selected by Isolate and Optiprep gradient, at the concentrations and small volumes used, demonstrated similar characteristics and in vitro embryo production to conventional Percoll † gradient. † , en petits volumes, sur la qualite´de la semence, la production in vitro d'embryons, le rapport des sexes et la survie de l'embryon apre`s vitrification. Des semences de´congele´es ont e´te´attribue´es ale´atoirement a`une de quatre me´thodes de se´lection par gradient de densite´: (1) Percoll † traditionnel (P), (2) MiniPercoll (MP), (3) MiniIsolate (MI), et (4) MiniOptiprep (MO). La cine´tique et la qualite´des spermatozoı¨des ont e´te´e´value´es. L'utilisation des gradients P, MP et MI n'ont eu aucun effet sur la motilite´des spermatozoı¨des (P!0,05). Il y avait, cependant, une diminution des motilite´s totales et progressives dans le gradient MO (70,8 et 51,3 % c. 87,3 et 69,5 % pour P; 87,3 et 73 % pour MP; 92,3 et 78,8 % pour MI; PB0,05). Le gradient MO avait une inte´grite´de membrane infe´rieure aux gradients P, MP et MI (39,7 c. 70,5; 72,3; 63,8 % respectivement, PB0,05). Le pourcentage de blastocystes produits e´tait plus e´leve´apre`s gradient MI qu'apre`s les gradients MP et MO (21,1 c. 16,1 et 16,9 %, P B0,05) et similaire apre`s gradient P (18,4 %; P!0,05). Le rapport des sexes et la survie de l'embryon apre`s vitrification e´taient similaires parmi tous les groupes (P !0,05). Les semences choisies par gradients Isolate et Optiprep, aux concentrations et aux petits volumes utilise´s, de´montrent des caracte´ristiques et une production in vitro d'embryons similaires a`celles choisies par gradients Percoll † traditionnels.

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Section: Vianna Et Al * Sperm Selection In Bovine Embryo Production 39supporting

confidence: 91%

Isolate^® and Optiprep^® minigradients as alternatives for sperm selection in bovine in vitro embryo production

et al. 2014

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“…Factors that have been reported to influence sex ratio in cattle are the timing of insemination in vivo [35], the maturation stage of the oocyte at the time of in vitro fertilization [15], modifications of the fertilization procedure [20,36], and the postfertilization culture conditions [37].…”

Section: Discussionmentioning

confidence: 99%

“…Thus, in a recent large field trial of AI using sexed spermatozoa from seven Holstein bulls, Frijters et al [17] reported a 13.6% decline in the 56-day no return rate; many works also reported decreases in blastocysts rates after IVF of oocytes with sex-sorted sperm [18,19]. Proposed factors for the reduced fertility rates in vivo include not only the damage to the spermatozoa caused by the sorting procedure [11], but also the lower doses of spermatozoa used [9], and sire effects [20] (for review, see Rath et al [21]). …”

Section: Introductionmentioning

confidence: 99%

In vitro and in vivo quality of bovine embryos in vitro produced with sex-sorted sperm

et al. 2012

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In this work we analyzed the effects of three culture systems on developmental ability of bovine embryos in vitro produced with sexed sperm, the survival to vitrification (cryologic vitrification method) of such blastocysts, and their pregnancy rates after embryo transfer to recipients, both as fresh and after vitrification/warming. Finally, we measured the accuracy of the sorting protocol by a polymerase chain reaction-based method to validate the embryo sex at blastocyst stages. We confirmed an individual effect of the bull as well as development rates of embryos produced with sorted sperm lower than embryos with unsorted sperm, independent of the culture system used. The cryoresistance to vitrification of embryos produced with sexed sperm did not differ from that of conventionally produced embryos (re-expansion rates at 24 and 48 h: 74.6% vs. 75.5%, and 64.5% vs. 68.1% for embryos produced with conventional and sorted sperm, respectively; hatching rates at 48 h: 63.55% vs. 55.5% for embryos produced with conventional and sorted sperm, respectively). Finally, no significant differences were found in pregnancy rates after the embryo transfer of fresh and vitrified/warmed blastocysts (52.8% vs. 42.0%, respectively; P Ͼ 0.05). Male and female embryos produced with sorted sperm showed the same quality in terms of developmental ability, cryoresistance, and pregnancy rates after transfer. Our culture system, coupled with the vitrification in fiber plugs, provides good quality sex-known embryos which survive vitrification at similar rates than embryos produced with conventional unsorted sperm; also it produces good pregnancy rates after transfer of sexed embryos both fresh and after vitrification and warming.

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“…Several authors observed higher percentage of cells with intact plasma membrane in Percoll centrifuged semen (LEE et al, 2009;MACHADO et al, 2009;OLIVEIRA et al, 2012b), indicating that most dead spermatozoa present in frozen-thawed semen were retained in the upper layers of the gradient. Damaged spermatozoa are removed by a particular mechanism in Percoll separation methods (OSHIO, 1988;RHEMREV et al, 1989) and sperm progressive motility is generally improved after Percoll centrifugation (CESARI et al, 2006;LEE et al, 2009;MACHADO et al, 2009;PETYIM et al, 2009). In the present study the small amount of semen (150 µL) being deposited in the upper layer of this density gradient might have avoided this phenomenon to happen.…”

Section: Resultsmentioning

confidence: 99%

“…Table 3 -Mean (± SD) of plasma membrane integrity (PMI; %), sperm morphology (major, minor and total defects; %), sperm concentration (CONC; x10 6 sptz/mL), and number of sperm presenting membrane integrity (PMI*CONC; x10 6 sptz/ mL) after different laboratory challenges using different batches from a Brangus bull -Sertãozinho, SP -January 2016 Percoll centrifugation is a widely used method for bovine sperm selection (MCCANN;CHANTLER, 2000;PETYIM et al, 2009) mainly before IVP (CESARI et al, 2006;LEE et al, 2009;MACHADO et al, 2009). Several authors observed higher percentage of cells with intact plasma membrane in Percoll centrifuged semen (LEE et al, 2009;MACHADO et al, 2009;OLIVEIRA et al, 2012b), indicating that most dead spermatozoa present in frozen-thawed semen were retained in the upper layers of the gradient. Damaged spermatozoa are removed by a particular mechanism in Percoll separation methods (OSHIO, 1988;RHEMREV et al, 1989) and sperm progressive motility is generally improved after Percoll centrifugation (CESARI et al, 2006;LEE et al, 2009;MACHADO et al, 2009;PETYIM et al, 2009).…”

Section: Resultsmentioning

confidence: 99%

Assessment of different in vitro sperm challenges and in vivo fertility of bovine semen batches

Oliveira

Ribeiro

Silva

et al. 2017

Braz. J. Vet. Res. Anim. Sci.

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The aim of this work was to submit sperm cells to different laboratory challenges and to compare in vitro results with in vivo semen fertility. Four different batches from the same Brangus bull were used in a timed-AI program of 332 Brangus cows. Each batch (B) was submitted to the following procedure: semen sample was thawed at 36°C for 30 seconds (control). Sperm motility parameters, plasma membrane integrity, sperm morphology, and concentration were assessed. Then, an aliquot of thawed sample was incubated in a water bath at 45°C for 40 min (thermal challenge group; TCG) and another aliquot was centrifuged at 500 xg (Percoll gradient 45%/90%) for 15 min (centrifugation challenge group; CCG). Centrifuged semen was also submitted to another thermal challenge, being incubated (water bath) at 45°C for 40 min (centrifugation + thermal challenge group; CTCG). At the end of each challenge (CCG, TCG, and CTCG), the same laboratory tests used for control group were repeated. The following conception rates (CR) were observed for each batch: B1 = 48.9% (44/90); B2 = 44.2% (23/52); B3 = 55.5% (40/72); B4 = 43.2% (51/118); (p < 0.10). In the lab, B3 presented higher (p ≤ 0.05) progressive motility (PM) than B4 after thawing (control group) and after all sperm challenges (TCG, CCG, and CTCG). However, despite B3 and B4 having demonstrated a similar percentage of plasma membrane integrity (PMI) to the control group (B3 = 66.7 ± 1.3 and B4 = 65.2 ± 3.3), B3 demonstrated higher (P ≤ 0.05) percentage of PMI (37.2 ± 2.5) than B4 (26.7 ± 3.3) after passing through the most stressing in vitro challenge (CTCG). The semen batch presenting the highest resistance to in vitro challenges was the one that presented a trend for higher in vivo fertility, suggesting that submitting semen samples to laboratory challenges may be an interesting alternative for selecting batches with greater field fertility. Keywords: Conception rate. In vitro sperm resistance. Timed-AI. ResumoO objetivo deste estudo foi estressar células espermáticas em diferentes desafios laboratoriais e comparar os resultados in vitro com a fertilidade in vivo do sêmen. Quatro partidas de um mesmo touro Brangus foram utilizadas em um programa de IATF de 332 vacas Brangus. Cada partida foi submetida ao seguinte procedimento: a amostra de sêmen foi descongelada a 36°C por 30 segundos (grupo controle). Foram avaliados parâmetros de motilidade espermática (CASA), integridade da membrana plasmática (PMI), morfologia e concentração espermática. Em seguida, uma alíquota da amostra descongelada foi incubada em banho-maria a 45°C durante 40 minutos (grupo de desafio térmico, TCG) e outra alíquota foi centrifugada a 500 xg (gradiente de Percoll 45%/90%) durante 15 min (grupo desafio de centrifugação, CCG). Uma aliquota do sêmen centrifugado foi ainda submetida ao desafio térmico, sendo incubado a 45°C durante 40 min (grupo de desafio térmico + centrifugação, CTCG). No final de cada desafio (CCG, TCG e CTCG), os mesmos testes laboratoriais utilizados para o grupo de controle f...

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Effect of Percoll volume, duration and force of centrifugation, on in vitro production and sex ratio of bovine embryos

Cited by 122 publications

References 37 publications

Isolate^® and Optiprep^® minigradients as alternatives for sperm selection in bovine in vitro embryo production

Isolate^® and Optiprep^® minigradients as alternatives for sperm selection in bovine in vitro embryo production

In vitro and in vivo quality of bovine embryos in vitro produced with sex-sorted sperm

Assessment of different in vitro sperm challenges and in vivo fertility of bovine semen batches

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Effect of Percoll volume, duration and force of centrifugation, on in vitro production and sex ratio of bovine embryos

Cited by 122 publications

References 37 publications

Isolate® and Optiprep® minigradients as alternatives for sperm selection in bovine in vitro embryo production

Isolate® and Optiprep® minigradients as alternatives for sperm selection in bovine in vitro embryo production

In vitro and in vivo quality of bovine embryos in vitro produced with sex-sorted sperm

Assessment of different in vitro sperm challenges and in vivo fertility of bovine semen batches

Contact Info

Product

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About

Isolate^® and Optiprep^® minigradients as alternatives for sperm selection in bovine in vitro embryo production

Isolate^® and Optiprep^® minigradients as alternatives for sperm selection in bovine in vitro embryo production