Generation of 2C-like mouse embryonic stem cells in vivo to evaluate developmental potency
Summary: This protocol describes an optimized workflow for generating 2-cell (2C) stage-like mouse embryonic stem cells (mESCs) for microinjection into 8-cell stage mouse embryos to evaluate the developmental potency of these cells. We detail the following steps: 1) chemical suppression of glycolysi...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English |
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Elsevier
2022-12-01
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| Series: | STAR Protocols |
| Subjects: | |
| Online Access: | http://www.sciencedirect.com/science/article/pii/S2666166722005640 |
| _version_ | 1798006850634907648 |
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| author | Dennis EK Tan Baojiang Wu Siqin Bao Wee-Wei Tee |
| author_facet | Dennis EK Tan Baojiang Wu Siqin Bao Wee-Wei Tee |
| author_sort | Dennis EK Tan |
| collection | DOAJ |
| description | Summary: This protocol describes an optimized workflow for generating 2-cell (2C) stage-like mouse embryonic stem cells (mESCs) for microinjection into 8-cell stage mouse embryos to evaluate the developmental potency of these cells. We detail the following steps: 1) chemical suppression of glycolysis to induce a 2C-like state in mESCs, 2) flow cytometry to enrich for 2C-like cells, 3) embryo microinjection of 2C-like mESCs into 8-cell stage mouse embryos, and finally, 4) immunofluorescence staining of the chimeric blastocysts.For complete details on the use and execution of this protocol, please refer to Hu et al. (2020). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. |
| first_indexed | 2024-04-11T13:02:17Z |
| format | Article |
| id | doaj.art-033270fd5ad140fba23019134910d1e5 |
| institution | Directory Open Access Journal |
| issn | 2666-1667 |
| language | English |
| last_indexed | 2024-04-11T13:02:17Z |
| publishDate | 2022-12-01 |
| publisher | Elsevier |
| record_format | Article |
| series | STAR Protocols |
| spelling | doaj.art-033270fd5ad140fba23019134910d1e52022-12-22T04:22:53ZengElsevierSTAR Protocols2666-16672022-12-0134101684Generation of 2C-like mouse embryonic stem cells in vivo to evaluate developmental potencyDennis EK Tan0Baojiang Wu1Siqin Bao2Wee-Wei Tee3Chromatin Dynamics and Disease Epigenetics Group, Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A∗STAR), Singapore, SingaporeResearch Centre for Animal Genetic Resources of Mongolia Plateau, College of Life Sciences, Inner Mongolia University, Hohhot, ChinaResearch Centre for Animal Genetic Resources of Mongolia Plateau, College of Life Sciences, Inner Mongolia University, Hohhot, ChinaChromatin Dynamics and Disease Epigenetics Group, Institute of Molecular and Cell Biology, Agency for Science, Technology and Research (A∗STAR), Singapore, Singapore; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore, Singapore; Corresponding authorSummary: This protocol describes an optimized workflow for generating 2-cell (2C) stage-like mouse embryonic stem cells (mESCs) for microinjection into 8-cell stage mouse embryos to evaluate the developmental potency of these cells. We detail the following steps: 1) chemical suppression of glycolysis to induce a 2C-like state in mESCs, 2) flow cytometry to enrich for 2C-like cells, 3) embryo microinjection of 2C-like mESCs into 8-cell stage mouse embryos, and finally, 4) immunofluorescence staining of the chimeric blastocysts.For complete details on the use and execution of this protocol, please refer to Hu et al. (2020). : Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics.http://www.sciencedirect.com/science/article/pii/S2666166722005640Flow cytometry/Mass cytometryDevelopmental biologyMicroscopyStem cellsCell differentiation |
| spellingShingle | Dennis EK Tan Baojiang Wu Siqin Bao Wee-Wei Tee Generation of 2C-like mouse embryonic stem cells in vivo to evaluate developmental potency STAR Protocols Flow cytometry/Mass cytometry Developmental biology Microscopy Stem cells Cell differentiation |
| title | Generation of 2C-like mouse embryonic stem cells in vivo to evaluate developmental potency |
| title_full | Generation of 2C-like mouse embryonic stem cells in vivo to evaluate developmental potency |
| title_fullStr | Generation of 2C-like mouse embryonic stem cells in vivo to evaluate developmental potency |
| title_full_unstemmed | Generation of 2C-like mouse embryonic stem cells in vivo to evaluate developmental potency |
| title_short | Generation of 2C-like mouse embryonic stem cells in vivo to evaluate developmental potency |
| title_sort | generation of 2c like mouse embryonic stem cells in vivo to evaluate developmental potency |
| topic | Flow cytometry/Mass cytometry Developmental biology Microscopy Stem cells Cell differentiation |
| url | http://www.sciencedirect.com/science/article/pii/S2666166722005640 |
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