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Improved blastocyst development of single cow OPU-derived presumptive zygotes by group culture with agarose-embedded helper embryos

BACKGROUND: The in vitro culture of presumed zygotes derived from single cow ovum pick-up (OPU) is important for the production of quality blastocysts maintaining pedigree. The aim of the present study was to evaluate the agar chip-embedded helper embryo coculture system for single cow OPU-derived z...

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Autores principales: Deb, Gautam Kumar, Jin, Jong In, Kwon, Tae Hyun, Choi, Byung Hyun, Bang, Jae Il, Dey, Shukla Rani, Cho, In Rae, Kong, Il Keun
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2011
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3177903/
https://www.ncbi.nlm.nih.gov/pubmed/21864328
http://dx.doi.org/10.1186/1477-7827-9-121
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author Deb, Gautam Kumar
Jin, Jong In
Kwon, Tae Hyun
Choi, Byung Hyun
Bang, Jae Il
Dey, Shukla Rani
Cho, In Rae
Kong, Il Keun
author_facet Deb, Gautam Kumar
Jin, Jong In
Kwon, Tae Hyun
Choi, Byung Hyun
Bang, Jae Il
Dey, Shukla Rani
Cho, In Rae
Kong, Il Keun
author_sort Deb, Gautam Kumar
collection PubMed
description BACKGROUND: The in vitro culture of presumed zygotes derived from single cow ovum pick-up (OPU) is important for the production of quality blastocysts maintaining pedigree. The aim of the present study was to evaluate the agar chip-embedded helper embryo coculture system for single cow OPU-derived zygotes by assessing embryo quality. METHODS: Cumulus oocyte complexes (COCs) were collected from Hanwoo cows with high genetic merit twice a week using the ultra-sound guided OPU technique and from slaughterhouse ovaries. The Hanwoo cow COCs and slaughterhouse ovaries were matured in vitro, fertilized in vitro with thawed Hanwoo sperm and cultured for 24 h. The presumed zygotes were subsequently placed in three different culture systems: (1) control OPU (controlOPU) with single cow OPU-derived presumed zygotes (2~8); (2) agar chip-embedded slaughterhouse helper embryo coculture (agarOPU) with ten presumed zygotes including all presumed zygotes from a cow (2~8) and the rest from agar chip-embedded slaughterhouse presumed zygotes (8~2); and (3) slaughterhouse in vitro embryo production (sIVP) with ten slaughterhouse ovary-derived presumed zygotes, each in 50 μL droplets. Day 8 blastocysts were assayed for apoptosis and gene expression using real time PCR. RESULTS: The coculture system promoted higher blastocyst development in OPU zygotes compared to control OPU zygotes cultured alone (35.2 vs. 13.9%; P < 0.01). Genes predicted to be involved in implantation failure and/or embryo resorption were down-regulated (P < 0.05) in control OPU zygotes (CD9, 0.4-fold; AKRAB1, 0.3-fold) and in cocultured zygotes (CD9, 0.3-fold; AKRAB1, 0.3-fold) compared to sIVP blastocysts (1.0-fold). Moreover, genes involved in implantation and/or normal calf delivery were up-regulated (P < 0.05 to P < 0.01) in control OPU zygotes (PGSH2, 5.0-fold; TXN, 4.3-fold; PLAU, 1.7-fold) and cocultured zygotes (PGSH2, 14.5-fold; TXN, 3.2-fold; PLAU, 6.8-fold) compared to sIVP (1.0-fold) blastocysts. However, the expression of PLAC8, TGF-β1, ODC1, ATP5A1 and CASP3 did not differ between the three culture groups. CONCLUSIONS: Results show that the agar chip-embedded helper embryo coculture system enhances developmental competence and embryo quality in cultures of limited numbers of high pedigree single cow OPU presumed zygotes.
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spelling pubmed-31779032011-09-22 Improved blastocyst development of single cow OPU-derived presumptive zygotes by group culture with agarose-embedded helper embryos Deb, Gautam Kumar Jin, Jong In Kwon, Tae Hyun Choi, Byung Hyun Bang, Jae Il Dey, Shukla Rani Cho, In Rae Kong, Il Keun Reprod Biol Endocrinol Research BACKGROUND: The in vitro culture of presumed zygotes derived from single cow ovum pick-up (OPU) is important for the production of quality blastocysts maintaining pedigree. The aim of the present study was to evaluate the agar chip-embedded helper embryo coculture system for single cow OPU-derived zygotes by assessing embryo quality. METHODS: Cumulus oocyte complexes (COCs) were collected from Hanwoo cows with high genetic merit twice a week using the ultra-sound guided OPU technique and from slaughterhouse ovaries. The Hanwoo cow COCs and slaughterhouse ovaries were matured in vitro, fertilized in vitro with thawed Hanwoo sperm and cultured for 24 h. The presumed zygotes were subsequently placed in three different culture systems: (1) control OPU (controlOPU) with single cow OPU-derived presumed zygotes (2~8); (2) agar chip-embedded slaughterhouse helper embryo coculture (agarOPU) with ten presumed zygotes including all presumed zygotes from a cow (2~8) and the rest from agar chip-embedded slaughterhouse presumed zygotes (8~2); and (3) slaughterhouse in vitro embryo production (sIVP) with ten slaughterhouse ovary-derived presumed zygotes, each in 50 μL droplets. Day 8 blastocysts were assayed for apoptosis and gene expression using real time PCR. RESULTS: The coculture system promoted higher blastocyst development in OPU zygotes compared to control OPU zygotes cultured alone (35.2 vs. 13.9%; P < 0.01). Genes predicted to be involved in implantation failure and/or embryo resorption were down-regulated (P < 0.05) in control OPU zygotes (CD9, 0.4-fold; AKRAB1, 0.3-fold) and in cocultured zygotes (CD9, 0.3-fold; AKRAB1, 0.3-fold) compared to sIVP blastocysts (1.0-fold). Moreover, genes involved in implantation and/or normal calf delivery were up-regulated (P < 0.05 to P < 0.01) in control OPU zygotes (PGSH2, 5.0-fold; TXN, 4.3-fold; PLAU, 1.7-fold) and cocultured zygotes (PGSH2, 14.5-fold; TXN, 3.2-fold; PLAU, 6.8-fold) compared to sIVP (1.0-fold) blastocysts. However, the expression of PLAC8, TGF-β1, ODC1, ATP5A1 and CASP3 did not differ between the three culture groups. CONCLUSIONS: Results show that the agar chip-embedded helper embryo coculture system enhances developmental competence and embryo quality in cultures of limited numbers of high pedigree single cow OPU presumed zygotes. BioMed Central 2011-08-24 /pmc/articles/PMC3177903/ /pubmed/21864328 http://dx.doi.org/10.1186/1477-7827-9-121 Text en Copyright ©2011 Deb et al; licensee BioMed Central Ltd. http://creativecommons.org/licenses/by/2.0 This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
spellingShingle Research
Deb, Gautam Kumar
Jin, Jong In
Kwon, Tae Hyun
Choi, Byung Hyun
Bang, Jae Il
Dey, Shukla Rani
Cho, In Rae
Kong, Il Keun
Improved blastocyst development of single cow OPU-derived presumptive zygotes by group culture with agarose-embedded helper embryos
title Improved blastocyst development of single cow OPU-derived presumptive zygotes by group culture with agarose-embedded helper embryos
title_full Improved blastocyst development of single cow OPU-derived presumptive zygotes by group culture with agarose-embedded helper embryos
title_fullStr Improved blastocyst development of single cow OPU-derived presumptive zygotes by group culture with agarose-embedded helper embryos
title_full_unstemmed Improved blastocyst development of single cow OPU-derived presumptive zygotes by group culture with agarose-embedded helper embryos
title_short Improved blastocyst development of single cow OPU-derived presumptive zygotes by group culture with agarose-embedded helper embryos
title_sort improved blastocyst development of single cow opu-derived presumptive zygotes by group culture with agarose-embedded helper embryos
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3177903/
https://www.ncbi.nlm.nih.gov/pubmed/21864328
http://dx.doi.org/10.1186/1477-7827-9-121
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