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In Vitro Fertilization using Non-sexed and Sexed Bovine Sperm: Sperm Concentration, Sorter Pressure, and Bull Effects

The objective of these experiments was to study bovine in vitro fertilization (IVF) conditions for blastocyst production using non-sexed sperm (Experiment 1) and sexed sperm (Experiment 2). For Experiment 1, in vitro-matured oocytes (N = 707) were allocated to a 2 × 3 × 4 factorial design: time of c...

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Published in:Reproduction in domestic animals 2011-06, Vol.46 (3), p.495-502
Main Authors: Barceló-Fimbres, M, Campos-Chillón, LF, Seidel, GE Jr
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creator Barceló-Fimbres, M
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description The objective of these experiments was to study bovine in vitro fertilization (IVF) conditions for blastocyst production using non-sexed sperm (Experiment 1) and sexed sperm (Experiment 2). For Experiment 1, in vitro-matured oocytes (N = 707) were allocated to a 2 × 3 × 4 factorial design: time of co-incubation of gametes for fertilization (4 and 18 h), sperm dose (1, 0.33, and 0.11 × 10⁶ frozen-thawed sperm/ml, and sperm source (four bulls). Pronuclear status was evaluated for a subset. Experiment 2 (N = 2155 oocytes) was a 2 × 3 × 2 × 6 factorial design: sex of sperm (X and Y), sperm dose (1, 0.33, and 0.11 × 10⁶ frozen-thawed sperm/ml), and sperm-sorting pressures (40 and 50 psi), replicated with sperm of six bulls. Presumptive zygotes were cultured 60 h in chemically defined medium-1 (CDM-1), and for 114 h in CDM-2. For Experiment 1, pronuclear formation, cleavage and blastocysts rates were greater for 1, and 0.33 × 10⁶ than 0.11 × 10⁶ sperm/ml (72 and 62 vs 42%; 89 and 81 vs 58%; and 21 and 17 vs 9%, respectively; all p < 0.01); polyspermy was greater for 1, than 0.33 and 0.11 × 10⁶ sperm concentrations (24 vs 2 and 0%; p < 0.01). There were greater main effects (p < 0.01) of pronuclear formation (69 vs 48%), polyspermy (13 vs 4%), and cleavage (63 vs 54%), at 18 than at 4 h of co-incubation of gametes (all p < 0.01). For Experiment 2, cleavage and blastocyst rates were greater for 1 × 10⁶ sperm/ml vs 0.33 and 0.11 (69%, 47%, and 30% cleavage and 30%, 14%, and 8% blastocysts) and 40 vs 50 psi (54% and 44% cleavage and 18% and 15% blastocysts) (p < 0.01). A marked bull by fertilization sperm dose interaction was found for cleavage (p < 0.05). The main conclusion was that the optimal sperm concentration for cleavage and producing blastocysts via IVF with sexed sperm was considerably higher and more variable among bulls than for unsexed sperm.
doi_str_mv 10.1111/j.1439-0531.2010.01696.x
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For Experiment 1, in vitro-matured oocytes (N = 707) were allocated to a 2 × 3 × 4 factorial design: time of co-incubation of gametes for fertilization (4 and 18 h), sperm dose (1, 0.33, and 0.11 × 10⁶ frozen-thawed sperm/ml, and sperm source (four bulls). Pronuclear status was evaluated for a subset. Experiment 2 (N = 2155 oocytes) was a 2 × 3 × 2 × 6 factorial design: sex of sperm (X and Y), sperm dose (1, 0.33, and 0.11 × 10⁶ frozen-thawed sperm/ml), and sperm-sorting pressures (40 and 50 psi), replicated with sperm of six bulls. Presumptive zygotes were cultured 60 h in chemically defined medium-1 (CDM-1), and for 114 h in CDM-2. For Experiment 1, pronuclear formation, cleavage and blastocysts rates were greater for 1, and 0.33 × 10⁶ than 0.11 × 10⁶ sperm/ml (72 and 62 vs 42%; 89 and 81 vs 58%; and 21 and 17 vs 9%, respectively; all p < 0.01); polyspermy was greater for 1, than 0.33 and 0.11 × 10⁶ sperm concentrations (24 vs 2 and 0%; p < 0.01). 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Psychology ; In vitro fertilization ; Male ; Males ; oocytes ; polyspermy ; Semen Preservation - veterinary ; Sex Preselection - veterinary ; Sperm Capacitation ; sperm concentration ; Sperm Count - veterinary ; spermatozoa ; Spermatozoa - classification ; Spermatozoa - cytology ; Terrestrial animal productions ; Vertebrates</subject><ispartof>Reproduction in domestic animals, 2011-06, Vol.46 (3), p.495-502</ispartof><rights>2010 Blackwell Verlag GmbH</rights><rights>2015 INIST-CNRS</rights><rights>2010 Blackwell Verlag GmbH.</rights><lds50>peer_reviewed</lds50><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c5776-3c25490d8a1a8c201499e7d1e9ad56dd6da1581fc2f74b8337bc1707a80576cd3</citedby><cites>FETCH-LOGICAL-c5776-3c25490d8a1a8c201499e7d1e9ad56dd6da1581fc2f74b8337bc1707a80576cd3</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><link.rule.ids>314,780,784,27924,27925</link.rule.ids><backlink>$$Uhttp://pascal-francis.inist.fr/vibad/index.php?action=getRecordDetail&amp;idt=24141967$$DView record in Pascal Francis$$Hfree_for_read</backlink><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/20946538$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><creatorcontrib>Barceló-Fimbres, M</creatorcontrib><creatorcontrib>Campos-Chillón, LF</creatorcontrib><creatorcontrib>Seidel, GE Jr</creatorcontrib><title>In Vitro Fertilization using Non-sexed and Sexed Bovine Sperm: Sperm Concentration, Sorter Pressure, and Bull Effects</title><title>Reproduction in domestic animals</title><addtitle>Reprod Domest Anim</addtitle><description><![CDATA[The objective of these experiments was to study bovine in vitro fertilization (IVF) conditions for blastocyst production using non-sexed sperm (Experiment 1) and sexed sperm (Experiment 2). For Experiment 1, in vitro-matured oocytes (N = 707) were allocated to a 2 × 3 × 4 factorial design: time of co-incubation of gametes for fertilization (4 and 18 h), sperm dose (1, 0.33, and 0.11 × 10⁶ frozen-thawed sperm/ml, and sperm source (four bulls). Pronuclear status was evaluated for a subset. Experiment 2 (N = 2155 oocytes) was a 2 × 3 × 2 × 6 factorial design: sex of sperm (X and Y), sperm dose (1, 0.33, and 0.11 × 10⁶ frozen-thawed sperm/ml), and sperm-sorting pressures (40 and 50 psi), replicated with sperm of six bulls. Presumptive zygotes were cultured 60 h in chemically defined medium-1 (CDM-1), and for 114 h in CDM-2. For Experiment 1, pronuclear formation, cleavage and blastocysts rates were greater for 1, and 0.33 × 10⁶ than 0.11 × 10⁶ sperm/ml (72 and 62 vs 42%; 89 and 81 vs 58%; and 21 and 17 vs 9%, respectively; all p < 0.01); polyspermy was greater for 1, than 0.33 and 0.11 × 10⁶ sperm concentrations (24 vs 2 and 0%; p < 0.01). 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Psychology</subject><subject>In vitro fertilization</subject><subject>Male</subject><subject>Males</subject><subject>oocytes</subject><subject>polyspermy</subject><subject>Semen Preservation - veterinary</subject><subject>Sex Preselection - veterinary</subject><subject>Sperm Capacitation</subject><subject>sperm concentration</subject><subject>Sperm Count - veterinary</subject><subject>spermatozoa</subject><subject>Spermatozoa - classification</subject><subject>Spermatozoa - cytology</subject><subject>Terrestrial animal productions</subject><subject>Vertebrates</subject><issn>0936-6768</issn><issn>1439-0531</issn><fulltext>true</fulltext><rsrctype>article</rsrctype><creationdate>2011</creationdate><recordtype>article</recordtype><recordid>eNqNkV9v0zAUxSMEYmXwFcBCQrwsxa7jf0g8bGUtQ2MgyuDRcm1nckntYifQ8elxmlIkJCT8ci37d67OvacoAIJjlM-L1RhVWJSQYDSewPwKERV0vL1TjA4fd4sRFJiWlFF-VDxIaQUhIpyx-8XRBIqKEsxHRXfhwWfXxgBmNraucT9V64IHXXL-BlwFXya7tQYob8BidzsL3523YLGxcf1yKGAavLa-jTvtCViE2NoIPkSbUhftyU591jUNOK9rq9v0sLhXqybZR_t6XFzPzj9N35SX7-cX09PLUhPGaIn1hFQCGq6Q4joPWglhmUFWKEOoMdSoPBGq9aRm1ZJjzJYaMcgUh4RRbfBx8Xzou4nhW2dTK9cuads0ytvQJckpQSJvhWXy6V_kKnTRZ3MZqjjCuKIZ4gOkY0gp2lpuolureCsRlH0wciX7_ct-_7IPRu6Ckdssfbzv3y3X1hyEv5PIwLM9oJJWTR2V1y794SpUIUF7o68G7odr7O1_G5AfX5_2t6wvB71Lrd0e9Cp-lbk7I_LL1VzOxezdWzSlsuefDHytglQ3MXu6XvRRQAgxJrvV_ZtAFDOIfwEXcska</recordid><startdate>201106</startdate><enddate>201106</enddate><creator>Barceló-Fimbres, M</creator><creator>Campos-Chillón, LF</creator><creator>Seidel, GE Jr</creator><general>Blackwell Publishing Ltd</general><general>Blackwell</general><scope>FBQ</scope><scope>BSCLL</scope><scope>IQODW</scope><scope>CGR</scope><scope>CUY</scope><scope>CVF</scope><scope>ECM</scope><scope>EIF</scope><scope>NPM</scope><scope>AAYXX</scope><scope>CITATION</scope><scope>7QO</scope><scope>8FD</scope><scope>FR3</scope><scope>P64</scope><scope>7X8</scope></search><sort><creationdate>201106</creationdate><title>In Vitro Fertilization using Non-sexed and Sexed Bovine Sperm: Sperm Concentration, Sorter Pressure, and Bull Effects</title><author>Barceló-Fimbres, M ; Campos-Chillón, LF ; Seidel, GE Jr</author></sort><facets><frbrtype>5</frbrtype><frbrgroupid>cdi_FETCH-LOGICAL-c5776-3c25490d8a1a8c201499e7d1e9ad56dd6da1581fc2f74b8337bc1707a80576cd3</frbrgroupid><rsrctype>articles</rsrctype><prefilter>articles</prefilter><language>eng</language><creationdate>2011</creationdate><topic>Animal productions</topic><topic>Animal reproduction</topic><topic>Animals</topic><topic>Biological and medical sciences</topic><topic>blastocyst</topic><topic>Blastocyst - physiology</topic><topic>bulls</topic><topic>Cattle</topic><topic>Cell Separation - veterinary</topic><topic>Cleavage Stage, Ovum - physiology</topic><topic>Cryopreservation - veterinary</topic><topic>Female</topic><topic>Fertilization in Vitro - veterinary</topic><topic>Fundamental and applied biological sciences. Psychology</topic><topic>In vitro fertilization</topic><topic>Male</topic><topic>Males</topic><topic>oocytes</topic><topic>polyspermy</topic><topic>Semen Preservation - veterinary</topic><topic>Sex Preselection - veterinary</topic><topic>Sperm Capacitation</topic><topic>sperm concentration</topic><topic>Sperm Count - veterinary</topic><topic>spermatozoa</topic><topic>Spermatozoa - classification</topic><topic>Spermatozoa - cytology</topic><topic>Terrestrial animal productions</topic><topic>Vertebrates</topic><toplevel>peer_reviewed</toplevel><toplevel>online_resources</toplevel><creatorcontrib>Barceló-Fimbres, M</creatorcontrib><creatorcontrib>Campos-Chillón, LF</creatorcontrib><creatorcontrib>Seidel, GE Jr</creatorcontrib><collection>AGRIS</collection><collection>Istex</collection><collection>Pascal-Francis</collection><collection>Medline</collection><collection>MEDLINE</collection><collection>MEDLINE (Ovid)</collection><collection>MEDLINE</collection><collection>MEDLINE</collection><collection>PubMed</collection><collection>CrossRef</collection><collection>Biotechnology Research Abstracts</collection><collection>Technology Research Database</collection><collection>Engineering Research Database</collection><collection>Biotechnology and BioEngineering Abstracts</collection><collection>MEDLINE - Academic</collection><jtitle>Reproduction in domestic animals</jtitle></facets><delivery><delcategory>Remote Search Resource</delcategory><fulltext>fulltext</fulltext></delivery><addata><au>Barceló-Fimbres, M</au><au>Campos-Chillón, LF</au><au>Seidel, GE Jr</au><format>journal</format><genre>article</genre><ristype>JOUR</ristype><atitle>In Vitro Fertilization using Non-sexed and Sexed Bovine Sperm: Sperm Concentration, Sorter Pressure, and Bull Effects</atitle><jtitle>Reproduction in domestic animals</jtitle><addtitle>Reprod Domest Anim</addtitle><date>2011-06</date><risdate>2011</risdate><volume>46</volume><issue>3</issue><spage>495</spage><epage>502</epage><pages>495-502</pages><issn>0936-6768</issn><eissn>1439-0531</eissn><abstract><![CDATA[The objective of these experiments was to study bovine in vitro fertilization (IVF) conditions for blastocyst production using non-sexed sperm (Experiment 1) and sexed sperm (Experiment 2). For Experiment 1, in vitro-matured oocytes (N = 707) were allocated to a 2 × 3 × 4 factorial design: time of co-incubation of gametes for fertilization (4 and 18 h), sperm dose (1, 0.33, and 0.11 × 10⁶ frozen-thawed sperm/ml, and sperm source (four bulls). Pronuclear status was evaluated for a subset. Experiment 2 (N = 2155 oocytes) was a 2 × 3 × 2 × 6 factorial design: sex of sperm (X and Y), sperm dose (1, 0.33, and 0.11 × 10⁶ frozen-thawed sperm/ml), and sperm-sorting pressures (40 and 50 psi), replicated with sperm of six bulls. Presumptive zygotes were cultured 60 h in chemically defined medium-1 (CDM-1), and for 114 h in CDM-2. For Experiment 1, pronuclear formation, cleavage and blastocysts rates were greater for 1, and 0.33 × 10⁶ than 0.11 × 10⁶ sperm/ml (72 and 62 vs 42%; 89 and 81 vs 58%; and 21 and 17 vs 9%, respectively; all p < 0.01); polyspermy was greater for 1, than 0.33 and 0.11 × 10⁶ sperm concentrations (24 vs 2 and 0%; p < 0.01). There were greater main effects (p < 0.01) of pronuclear formation (69 vs 48%), polyspermy (13 vs 4%), and cleavage (63 vs 54%), at 18 than at 4 h of co-incubation of gametes (all p < 0.01). For Experiment 2, cleavage and blastocyst rates were greater for 1 × 10⁶ sperm/ml vs 0.33 and 0.11 (69%, 47%, and 30% cleavage and 30%, 14%, and 8% blastocysts) and 40 vs 50 psi (54% and 44% cleavage and 18% and 15% blastocysts) (p < 0.01). A marked bull by fertilization sperm dose interaction was found for cleavage (p < 0.05). The main conclusion was that the optimal sperm concentration for cleavage and producing blastocysts via IVF with sexed sperm was considerably higher and more variable among bulls than for unsexed sperm.]]></abstract><cop>Oxford, UK</cop><pub>Blackwell Publishing Ltd</pub><pmid>20946538</pmid><doi>10.1111/j.1439-0531.2010.01696.x</doi><tpages>8</tpages></addata></record>
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identifier ISSN: 0936-6768
ispartof Reproduction in domestic animals, 2011-06, Vol.46 (3), p.495-502
issn 0936-6768
1439-0531
language eng
recordid cdi_proquest_miscellaneous_865190157
source Wiley
subjects Animal productions
Animal reproduction
Animals
Biological and medical sciences
blastocyst
Blastocyst - physiology
bulls
Cattle
Cell Separation - veterinary
Cleavage Stage, Ovum - physiology
Cryopreservation - veterinary
Female
Fertilization in Vitro - veterinary
Fundamental and applied biological sciences. Psychology
In vitro fertilization
Male
Males
oocytes
polyspermy
Semen Preservation - veterinary
Sex Preselection - veterinary
Sperm Capacitation
sperm concentration
Sperm Count - veterinary
spermatozoa
Spermatozoa - classification
Spermatozoa - cytology
Terrestrial animal productions
Vertebrates
title In Vitro Fertilization using Non-sexed and Sexed Bovine Sperm: Sperm Concentration, Sorter Pressure, and Bull Effects
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