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25 - In vitro culture and differentiation of mouse embryonic stem cells

Published online by Cambridge University Press:  11 August 2009

A. Rolletschek
Affiliation:
In Vitro Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany
C. Wiese
Affiliation:
Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany
A. M. Wobus
Affiliation:
Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany
Manuel Marí-Beffa
Affiliation:
Universidad de Málaga, Spain
Jennifer Knight
Affiliation:
University of Colorado, Boulder
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Summary

OBJECTIVE OF THE EXPERIMENT The aim of the experiments is (1) to establish mouse embryonic stem (ES) cell lines and (2) to study the differentiation of ES cells into cardiac and neuronal cells.

DEGREE OF DIFFICULTY Determination of alkaline phosphatase activity: easy. Immunofluorescence staining: moderate. All experiments concerning the establishment of ES cell lines and in vitro differentiation: difficult.

INTRODUCTION

Embryonic stem (ES) cells have been established as undifferentiated cell lines from the inner cell mass (ICM) of mouse and human blastocysts (Evans and Kaufman, 1981; Thomson et al., 1998). These pluripotent ES cells are characterized by a high proliferative capacity and the ability to develop into terminally differentiated cells of all three primary germ layers, the endodermal, ectodermal and mesodermal lineage (Figure 25.1). Mouse and human ES cells can differentiate in vitro to give rise to various somatic cell types (reviewed in Schuldiner et al., 2000; Wobus, 2001). In vivo, after transfer into blastocysts or by aggregation with blastomeres, mouse ES cells participate in the development of the embryo including the germ line (Bradley et al., 1984), whereas human ES cells after transplantation into nude mice give rise to teratomas (Thomson et al., 1998).

Besides ES cells, two other types of pluripotent embryonic cell types have been established as permanent lines: embryonic carcinoma (EC) cells derived from teratocarcinomas and embryonic germ (EG) cells isolated from primordial germ cells of early embryos (Stewart et al., 1994; Shamblott et al., 1998; see Figure 25.1).

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Publisher: Cambridge University Press
Print publication year: 2005

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References

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  • In vitro culture and differentiation of mouse embryonic stem cells
    • By A. Rolletschek, In Vitro Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany, C. Wiese, Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany, A. M. Wobus, Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany
  • Edited by Manuel Marí-Beffa, Universidad de Málaga, Spain, Jennifer Knight, University of Colorado, Boulder
  • Book: Key Experiments in Practical Developmental Biology
  • Online publication: 11 August 2009
  • Chapter DOI: https://doi.org/10.1017/CBO9780511546204.027
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  • In vitro culture and differentiation of mouse embryonic stem cells
    • By A. Rolletschek, In Vitro Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany, C. Wiese, Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany, A. M. Wobus, Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany
  • Edited by Manuel Marí-Beffa, Universidad de Málaga, Spain, Jennifer Knight, University of Colorado, Boulder
  • Book: Key Experiments in Practical Developmental Biology
  • Online publication: 11 August 2009
  • Chapter DOI: https://doi.org/10.1017/CBO9780511546204.027
Available formats
×

Save book to Google Drive

To save content items to your account, please confirm that you agree to abide by our usage policies. If this is the first time you use this feature, you will be asked to authorise Cambridge Core to connect with your account. Find out more about saving content to Google Drive.

  • In vitro culture and differentiation of mouse embryonic stem cells
    • By A. Rolletschek, In Vitro Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany, C. Wiese, Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany, A. M. Wobus, Differentiation Group, Dept. of Cytogenetics, Institute of Plant Genetics and Crop Plant Research (IPK), Corrensstr. 3, D-06466 Gatersleben, Germany
  • Edited by Manuel Marí-Beffa, Universidad de Málaga, Spain, Jennifer Knight, University of Colorado, Boulder
  • Book: Key Experiments in Practical Developmental Biology
  • Online publication: 11 August 2009
  • Chapter DOI: https://doi.org/10.1017/CBO9780511546204.027
Available formats
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