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Deriving and propagating mouse embryonic stem cell lines for studying genomic imprinting
1Division of Biology, Beckman Research Institute of the City of Hope, CA, USA.
Methods in Molecular Biology (Clifton, N.J.)
|July 5, 2003
Summary
Embryonic stem cells (ES cells) are derived from the blastocyst inner cell mass (ICM). While similar to ICM cells, ES cells are uniquely maintained in an undifferentiated state by factors like Leukemia inhibitory factor (LIF).
Area of Science:
- Developmental Biology
- Stem Cell Biology
- Cell Culture
Background:
- Embryonic stem (ES) cells originate from the blastocyst's inner cell mass (ICM).
- ES cells contribute to primitive ectoderm and endoderm derivatives, similar to early ICM cells.
- ES cells are not identical to ICM cells due to their unique culture environment.
Purpose of the Study:
- To investigate the nature of embryonic stem cells.
- To understand the factors maintaining ES cell pluripotency.
- To explore the relationship between ES cells and in vivo ICM cells.
Main Methods:
- Blastocyst injection chimera experiments.
- Cell culture techniques for ES cell propagation.
- Investigation of exogenous factors influencing ES cell division.
Main Results:
- ES cells contribute to ectoderm and endoderm lineages.
- ES cells are maintained in an undifferentiated state by external factors.
- Leukemia inhibitory factor (LIF) is crucial for mouse ES cell propagation on feeder layers.
Conclusions:
- ES cells are distinct from native ICM cells due to culture-induced modifications.
- Exogenous factors, such as LIF, lock ES cells in a continuous division cycle.
- LIF is essential for maintaining mouse ES cells, particularly when using primary embryo fibroblasts (PEFs) as feeder layers.