Germ-cell culture conditions facilitate the production of mouse embryonic stem cells
The derivation of embryonic stem-cell (ESC) lines from blastocysts is a very inefficient process. Murine ESCs are thought to arise from epiblast cells that are already predisposed to a primordial-germ-cell fate. During the process of ESC derivation from B6D2 F1 hybrid mice, if we first culture the embryo from the two-cell stage in medium supplemented with LIF, we improve the quality of the blastocyst. When the blastocyst is then cultured in a germ-line stem-cell culture medium (GSCm), we are able to more efficiently (28.3%) obtain quality ESC lines that have a normal karyotype, proper degree of chimerism, and exhibit germ-line transmission when microinjected into blastocysts. Although germ-cell-specific genes were expressed in all culture medium conditions, GSCm did not shift the transcriptome towards germ-cell specification. A correlation was further observed between ESC derivation efficiency and the expression of some imprinted genes and retrotransposable elements. In conclusion, the combination of LIF supplementation followed by culture in GSCm establishes a higher efficiency method for ESC derivation. © 2014 Wiley Periodicals, Inc.
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dig-inia-es-10261-2905522023-02-17T12:26:50Z Germ-cell culture conditions facilitate the production of mouse embryonic stem cells Ramos Ibeas, Priscila Pericuesta Camacho, Eva Fernández González, Raúl Gutiérrez Adán, Alfonso Ramírez De Paz, Miguel Ángel The derivation of embryonic stem-cell (ESC) lines from blastocysts is a very inefficient process. Murine ESCs are thought to arise from epiblast cells that are already predisposed to a primordial-germ-cell fate. During the process of ESC derivation from B6D2 F1 hybrid mice, if we first culture the embryo from the two-cell stage in medium supplemented with LIF, we improve the quality of the blastocyst. When the blastocyst is then cultured in a germ-line stem-cell culture medium (GSCm), we are able to more efficiently (28.3%) obtain quality ESC lines that have a normal karyotype, proper degree of chimerism, and exhibit germ-line transmission when microinjected into blastocysts. Although germ-cell-specific genes were expressed in all culture medium conditions, GSCm did not shift the transcriptome towards germ-cell specification. A correlation was further observed between ESC derivation efficiency and the expression of some imprinted genes and retrotransposable elements. In conclusion, the combination of LIF supplementation followed by culture in GSCm establishes a higher efficiency method for ESC derivation. © 2014 Wiley Periodicals, Inc. 2023-02-17T12:26:50Z 2023-02-17T12:26:50Z 2014 artículo Molecular Reproduction and Development 81: 794-804 (2014) 1040-452X http://hdl.handle.net/20.500.12792/3656 http://hdl.handle.net/10261/290552 10.1002/mrd.22346 1098-2795 en none Wiley |
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The derivation of embryonic stem-cell (ESC) lines from blastocysts is a very inefficient process. Murine ESCs are thought to arise from epiblast cells that are already predisposed to a primordial-germ-cell fate. During the process of ESC derivation from B6D2 F1 hybrid mice, if we first culture the embryo from the two-cell stage in medium supplemented with LIF, we improve the quality of the blastocyst. When the blastocyst is then cultured in a germ-line stem-cell culture medium (GSCm), we are able to more efficiently (28.3%) obtain quality ESC lines that have a normal karyotype, proper degree of chimerism, and exhibit germ-line transmission when microinjected into blastocysts. Although germ-cell-specific genes were expressed in all culture medium conditions, GSCm did not shift the transcriptome towards germ-cell specification. A correlation was further observed between ESC derivation efficiency and the expression of some imprinted genes and retrotransposable elements. In conclusion, the combination of LIF supplementation followed by culture in GSCm establishes a higher efficiency method for ESC derivation. © 2014 Wiley Periodicals, Inc. |
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Ramos Ibeas, Priscila Pericuesta Camacho, Eva Fernández González, Raúl Gutiérrez Adán, Alfonso Ramírez De Paz, Miguel Ángel |
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Ramos Ibeas, Priscila Pericuesta Camacho, Eva Fernández González, Raúl Gutiérrez Adán, Alfonso Ramírez De Paz, Miguel Ángel Germ-cell culture conditions facilitate the production of mouse embryonic stem cells |
author_facet |
Ramos Ibeas, Priscila Pericuesta Camacho, Eva Fernández González, Raúl Gutiérrez Adán, Alfonso Ramírez De Paz, Miguel Ángel |
author_sort |
Ramos Ibeas, Priscila |
title |
Germ-cell culture conditions facilitate the production of mouse embryonic stem cells |
title_short |
Germ-cell culture conditions facilitate the production of mouse embryonic stem cells |
title_full |
Germ-cell culture conditions facilitate the production of mouse embryonic stem cells |
title_fullStr |
Germ-cell culture conditions facilitate the production of mouse embryonic stem cells |
title_full_unstemmed |
Germ-cell culture conditions facilitate the production of mouse embryonic stem cells |
title_sort |
germ-cell culture conditions facilitate the production of mouse embryonic stem cells |
publisher |
Wiley |
publishDate |
2014 |
url |
http://hdl.handle.net/20.500.12792/3656 http://hdl.handle.net/10261/290552 |
work_keys_str_mv |
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