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The Importance of Mouse ES Cell Line Selection

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Advanced Protocols for Animal Transgenesis

Part of the book series: Springer Protocols Handbooks ((SPH))

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Abstract

The choice of a specific parental ES cell line used to create a genetically modified mouse has critical impact on the overall success of the project – affecting costs, complexity of effort, and time to efficient project completion. Despite the importance of making a thoughtful choice, many people default to employing a familiar cell line previously used in their laboratory or they are limited to the cell lines available from the service facility at their Institution. With careful consideration of a few key experimental parameters, the investigator can easily avoid unnecessary mistakes.

Historically, substrains of 129 strain mice have been used to create the majority of parental ES cell lines now in common use. Often these cell lines have been expanded many times and shared among colleagues, rather than being obtained directly from the laboratory in which they were originally established. Although this was a common practice in the early days of ES cell technology, avenues are now in place to assure optimal cell line history, health, genetic integrity, and performance. Methods for establishing new ES cell lines have greatly improved. Well-validated ES cell lines are now commercially available from a variety of genetic backgrounds beyond the 129-strain. In this chapter, we will discuss critical factors to consider, when choosing an ES cell line for a project.

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Abbreviations

ES cells:

Embryonic stem cells

MEF:

Mouse embryonic fibroblasts

BRL cells:

Buffalo rat liver cells

LIF:

Leukemia inhibitory factor

FBS:

Fetal bovine serum

KOSR:

Knockout serum replacement

BMP4:

Bone morphogenetic protein 4

GSK3:

Glycogen synthase kinase 3

MEK:

MAP kinase

FGF:

Fibroblast growth factor

GLT:

Germline transmission

ATCC:

American Type Culture Collection

BAC:

Bacterial artificial chromosome

SW:

Swiss Webster

DMEM:

Dulbecco’s Modified Eagle Medium

IKMC:

International Knockout Mouse Consortium

KOMP:

NIH Knockout Mouse Project

EUCOMM:

European Union Conditional Mouse Mutagenesis Program

NorCOMM:

North American Conditional Mouse Mutagenesis

NEAA:

Non-essential amino acids

Pen-Strep:

Penicillin and streptomycin

PE:

Plating efficiency

TC:

Tissue culture

TE:

Targeting efficiency

EP:

Electroporation

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Authors and Affiliations

  1. Velocigene, Regeneron Pharmaceutical Inc., 777 Old Saw Mill River Road, Terrytown, NY, 10591, USA

    Wojtek Auerbach

  2. IVP, OSI Pharmaceuticals, 1 Bioscience Park Drive, Farmingdale, NY, 11735, USA

    Anna B. Auerbach

Authors
  1. Wojtek Auerbach
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  2. Anna B. Auerbach
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Correspondence to Wojtek Auerbach .

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Editors and Affiliations

  1. Div. Biology, California Institute of Technology, Pasadena, 91125, California, USA

    Shirley Pease

  2. Transgenic Animal Model Core, University of Michigan, Ann Arbor, 48109-5674, Michigan, USA

    Thomas L. Saunders

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Auerbach, W., Auerbach, A.B. (2011). The Importance of Mouse ES Cell Line Selection. In: Pease, S., Saunders, T. (eds) Advanced Protocols for Animal Transgenesis. Springer Protocols Handbooks. Springer, Berlin, Heidelberg. https://doi.org/10.1007/978-3-642-20792-1_15

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  • DOI: https://doi.org/10.1007/978-3-642-20792-1_15

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