High pressure flow cytometric sorting damages sperm

Suh, T.K.; Schenk, J.L.; Seidel, G. E.

doi:10.1016/j.theriogenology.2005.02.002

Cited by 149 publications

(100 citation statements)

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“…Moreover, the sorting process is costly and has negative effects on quality of semen. Injuries have been reported because of the passage of semen by flow cytometry and laser incidence in DNA [4,5].…”

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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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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“…Normal operating pressure is 40 psi. Higher pressures (50 psi) have been attributed to increased sperm mortality and membrane impairment as well as lowered fertility and decreased rates of IVF (Suh et al, 2005;Schenk et al, 2009;Barceló-Fimbres et al 2010). Improvements have also been made in the electronics of the system.…”

Section: Historymentioning

confidence: 99%

Removing Seminal Plasma Improves Sex-Sorting of Bovine Sperm.

Burroughs

Graham

Lenz

et al. 2011

Biology of Reproduction

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REMOVING SEMINAL PLASMA IMPROVES SEX-SORTING OF BOVINE SPERMExperiments for this thesis evaluated how characteristics of bovine ejaculates affect efficiency of sorting X-from Y-chromosome bearing sperm by flow cytometry/cell sorting, and further, document that removal of seminal plasma improves efficacy of sexsorting bovine sperm. In Experiment I, ejaculates were collected by artificial vagina, two each from 10 bulls with an average of one hour between collections. Semen was centrifuged to separate sperm from seminal plasma, and then sperm were re-diluted to 160x10 6 sperm per ml with staining TALP (Tyrode's albumin, lactate, pyruvate) and 0, 5, 10 or 20% seminal plasma from the same ejaculate or reciprocally from first/second ejaculates. Following incubation with Hoechst 33342, sperm were sorted and analyzed with a flow cytometer/cell sorter. The % live-oriented sperm was higher for treatments with 0% seminal plasma (64.4%) than 5 (59.6%), 10 (59.0%) and 20% (57.8%) seminal plasma (p<0.01). The % live-oriented sperm was higher for second (63.0%) than first ejaculates (56.2%). Sort rate was higher for samples with 0% seminal plasma (p<0.05).The percentages of membrane-impaired sperm were lower for 0% (16.5%) than 5 (21.9%), 10 (23.6%) or 20% (23.4%) seminal plasma (p<0.003), and for second iii ejaculates (18.2%) compared to first ejaculates (25.9%). Whether seminal plasma was from first versus second ejaculates had no effect (p>0.1). Effects of seminal plasma originating from bulls different from those whose sperm were sorted was evaluated in Experiment II. Semen collection and initial analysis were performed as in Experiment I and for all successive experiments. Seminal plasma from ejaculates of 6 bulls (3 that sorted well; 3 that sorted poorly) used in Experiment I was used with sperm collected for Experiment II as well as seminal plasma from sperm donors. Sperm were stained with Hoechst 33342 as in Experiment I, but with 10% seminal plasma in staining TALP.Only seminal plasma from one bull resulted in differences for any sorting parameters; 31% of sperm were membrane-impaired with seminal plasma from this bull compared to 16-19% for seminal plasma from other bulls (p<0.05). Sort rate was decreased (3.02x10 3 sperm per sec) compared to a range of 3.64-3.97x10 3 sperm per sec with the other seminal plasmas (p<0.05). This may have been due to contamination of the original semen sample, since this effect was only seen with one of six bulls.Experiment III investigated effects of adding bovine serum albumin (BSA) during staining with Hoechst 33342 using semen from 10 bulls. BSA was added at 0, 0.3 or 0.9% to staining TALP with either 0 or 10% seminal plasma. Although BSA has been shown to be beneficial to sperm, there was a possibility that BSA would bind Hoechst 33342, thereby reducing sorting efficiency. There was no evidence of either effect; however, sperm with 0% seminal plasma had higher % live oriented cells (65%) than sperm incubated with 10% seminal plasma (61%; p<0.05). In addition, samples conta...

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“…A diminuição do sistema de pressão de 50 para 40 psi proporcionou uma melhora na qualidade do sêmen, tais como na motilidade, espermatozóides vivos com membrana intacta, um aumento na clivagem e desenvolvimento blastocístico sem diminuir o desempenho das máquinas (CAMPOS- CHILLON;DE LA TORRE, 2003;SUH et al, 2005). Por outro lado Suh et al…”

Section: Resultados Obtidos Com O Uso De Espermatozóides Sexadosunclassified

“…Estes resultados expõem a idéia de que os espermatozóides sexados são afetados por algum procedimento dentro do processo da sexagem e/ou congelação. Com esta teoria vários trabalhos foram realizados com o intuito de avaliar a viabilidade dos espermatozóides sexados após descongelação (PENFOLD et al, 1998;SUH et al, 2005;BOE-HANSON et al, 2005;MOCE et al, 2006;TANNO, 2009) com resultados variádos.…”

Section: N T R O D U ç ã O | 24unclassified

“…Porém com esta metodologia somente 20 a 30% dos espermatozóides poderia ser orientados . Em 1993 MAXWELL et al, 2004;SUH et al, 2005). O laser UV (Vanguard TM ) é um sistema de estado sólido diode-pumped (Newport Corporation, Irvine, CA, USA) que evita os efeitos lesivos de UV de baixo comprimento de onda que são absorvidos pelos ácidos nucléicos e proteínas (SEIDEL; GARNER, 2002).…”

Section: I T E R a T U R A | 39unclassified

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Avaliação das características da motilidade (CASA), morfologia e funcionalidade da membrana plasmática (HOST) de espermatozóides bovinos sexados por cimetria de fluxo

Gallego¹

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A verdade é que, quando sua mente estava completamente alheia, ele tinha o pensamento mais estranho que qualquer lunático no mundo jamais tivera, pois lhe parecia razoável e necessário, tanto por uma questão de honra quanto para servir a nação, se tornar um cavaleiro errante e viajar pelo mundo em sua armadura e seu cavalo para buscar aventuras e envolver-se em tudo que ele havia lido sobre como os cavaleiros errantes se envolviam, aproveitando as oportunidades, colocando-se em perigo e corrigindo todo tipo de mal.-Miguel de Cervantes, Don Quixote. DEDICO.A Dios.A mis Papas Ramiro y Alba Lucia, su amor es la base de todas mis logros, Dios me bendijo al poder tenerlos a mi lado.A mi hermanita María del Pilar, eres la otra faz de una obra de arte, tu alegría y emotividad me llenan de felicidad. No te imaginas como te admiro.A mis abuelitas Ruth y Mary, Las abuelitas más dulces y tiernas que puedan existir, siempre las llevo en mi corazón.A mis Abuelos Libardo y Jorge (In memoriam), Sus vidas son una inspiración para mi. Es un honor llevar el apellido Mejia Gallego. Los Extraño A mi novia Dunia, Porque nuestros sueños se hagan realidad. LOS AMO !!!!! Agradecimientos.A mis tias Angela, Nohora y Liliana: Siempre han creido en mis capacidades, ello me ha llenado de confianza para llegar a donde estoy ahora, de todas he aprendido cosas importantes en la vida.Las llevo y las llevare en mi corazón siempre.A mis tios

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High pressure flow cytometric sorting damages sperm

Cited by 149 publications

References 20 publications

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

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

Removing Seminal Plasma Improves Sex-Sorting of Bovine Sperm.

Avaliação das características da motilidade (CASA), morfologia e funcionalidade da membrana plasmática (HOST) de espermatozóides bovinos sexados por cimetria de fluxo

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