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Systematic analysis of the factors that adversely affect the rate of cell accumulation in mouse embryos during their culture in vitro

BACKGROUND: Retarded embryo growth is a pervasive effect of culture in vitro. METHODS: A systematic analysis of the interactions between media design, embryo culture density, oxygen tension, amino acids, trophic ligands and the genetic background of the mouse on embryo growth rates in vitro was perf...

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Autores principales: Jin, Xing L, O’Neill, Chris
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4036297/
https://www.ncbi.nlm.nih.gov/pubmed/24885989
http://dx.doi.org/10.1186/1477-7827-12-35
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author Jin, Xing L
O’Neill, Chris
author_facet Jin, Xing L
O’Neill, Chris
author_sort Jin, Xing L
collection PubMed
description BACKGROUND: Retarded embryo growth is a pervasive effect of culture in vitro. METHODS: A systematic analysis of the interactions between media design, embryo culture density, oxygen tension, amino acids, trophic ligands and the genetic background of the mouse on embryo growth rates in vitro was performed. RESULTS: Growth retardation of mouse zygotes was greater in 20% O(2) than 5%, a sequential media design was superior to static simple media designs, but the supplementation of simple media with mixed amino acids mitigated this difference. There was a beneficial effect of communal culture in small volumes, and supplementation with a trophic ligand (Paf) further enhanced growth rates. For hybrid strain zygotes (B6CBF1) communal culture in KSOM media supplemented with amino acids, albumin and Paf under 5% O(2) resulted in complete rescue of their rate of accumulation of cells and blastocyst formation. Inbred strain (C57BL6/J) zygotes, however, still showed some retardation of development under these conditions. The additional supplementation of media with another trophic ligand (IGF1) showed a further additive beneficial effect on development of inbred strain embryos but they still showed a growth deficit of ~ 23% cell number. The results show that optimising the interactions between a range of culture conditions and media design can rescue hybrid strain embryos from a retarded rate of cell proliferation caused by culture in vitro, but this was incomplete for the B6 strain. CONCLUSIONS: The results indicate that the growth requirement of embryos in vitro varies depending upon their genetic background and provide models for the further genetic analysis of embryo growth.
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spelling pubmed-40362972014-05-29 Systematic analysis of the factors that adversely affect the rate of cell accumulation in mouse embryos during their culture in vitro Jin, Xing L O’Neill, Chris Reprod Biol Endocrinol Research BACKGROUND: Retarded embryo growth is a pervasive effect of culture in vitro. METHODS: A systematic analysis of the interactions between media design, embryo culture density, oxygen tension, amino acids, trophic ligands and the genetic background of the mouse on embryo growth rates in vitro was performed. RESULTS: Growth retardation of mouse zygotes was greater in 20% O(2) than 5%, a sequential media design was superior to static simple media designs, but the supplementation of simple media with mixed amino acids mitigated this difference. There was a beneficial effect of communal culture in small volumes, and supplementation with a trophic ligand (Paf) further enhanced growth rates. For hybrid strain zygotes (B6CBF1) communal culture in KSOM media supplemented with amino acids, albumin and Paf under 5% O(2) resulted in complete rescue of their rate of accumulation of cells and blastocyst formation. Inbred strain (C57BL6/J) zygotes, however, still showed some retardation of development under these conditions. The additional supplementation of media with another trophic ligand (IGF1) showed a further additive beneficial effect on development of inbred strain embryos but they still showed a growth deficit of ~ 23% cell number. The results show that optimising the interactions between a range of culture conditions and media design can rescue hybrid strain embryos from a retarded rate of cell proliferation caused by culture in vitro, but this was incomplete for the B6 strain. CONCLUSIONS: The results indicate that the growth requirement of embryos in vitro varies depending upon their genetic background and provide models for the further genetic analysis of embryo growth. BioMed Central 2014-05-08 /pmc/articles/PMC4036297/ /pubmed/24885989 http://dx.doi.org/10.1186/1477-7827-12-35 Text en Copyright © 2014 Jin and O’Neill; 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 credited. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research
Jin, Xing L
O’Neill, Chris
Systematic analysis of the factors that adversely affect the rate of cell accumulation in mouse embryos during their culture in vitro
title Systematic analysis of the factors that adversely affect the rate of cell accumulation in mouse embryos during their culture in vitro
title_full Systematic analysis of the factors that adversely affect the rate of cell accumulation in mouse embryos during their culture in vitro
title_fullStr Systematic analysis of the factors that adversely affect the rate of cell accumulation in mouse embryos during their culture in vitro
title_full_unstemmed Systematic analysis of the factors that adversely affect the rate of cell accumulation in mouse embryos during their culture in vitro
title_short Systematic analysis of the factors that adversely affect the rate of cell accumulation in mouse embryos during their culture in vitro
title_sort systematic analysis of the factors that adversely affect the rate of cell accumulation in mouse embryos during their culture in vitro
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4036297/
https://www.ncbi.nlm.nih.gov/pubmed/24885989
http://dx.doi.org/10.1186/1477-7827-12-35
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