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Derivation of Stem Cell Lines from Mouse Preimplantation Embryos
Published on: August 20, 2017
A simple procedure for the efficient derivation of mouse ES cells
Esther S M Wong1, Kenneth H Ban, Rafidah Mutalif
1Institute of Medical Biology, Immunos, Singapore.
Methods in Enzymology
|August 10, 2010
Summary
Researchers developed an efficient method to derive mouse embryonic stem (ES) cells using defined serum-free media. This new technique improves the generation of genetically modified mice for studying gene function.
Area of Science:
- Stem Cell Biology
- Genetics
- Developmental Biology
Background:
- Embryonic stem (ES) cells are crucial for genetic modification in mice via gene targeting.
- Existing methods for deriving ES cell lines are often inefficient.
- There is a continuous need for new ES lines, especially from mutant mouse models, to study gene function.
Purpose of the Study:
- To develop a reproducible, efficient, and economical method for deriving mouse ES cells.
- To establish a protocol for routine maintenance and karyotyping of ES cells.
Main Methods:
- Derivation of ES cells from preimplantation stage mouse blastocysts.
- Utilized a defined serum-free, serum replacement (KO-SR) media.
- Established procedures for routine maintenance and karyotyping of derived ES cells.
Main Results:
- Achieved 50-85% efficiency in deriving ES cell lines from various mouse strains.
- Successfully derived over 100 ES lines, with >70% being euploid after karyotyping.
- Two derived ES lines demonstrated germ-line transmission in chimera experiments.
Conclusions:
- The described method offers a highly efficient and reproducible approach for mouse ES cell derivation.
- This protocol facilitates the generation of genetically modified mouse models for functional genomics research.
- The method is economical and suitable for routine use in stem cell laboratories.
