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Preparation of Mouse Embryonic Fibroblast Cells Suitable for Culturing Human Embryonic and Induced Pluripotent Stem Cells
Published on: June 21, 2012
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Isolation of Mouse Embryo Fibroblasts
Marian E Durkin1, Xiaolan Qian2, Nicholas C Popescu2
1Laboratory of Experimental Carcinogenesis, National Cancer Institute, NIH, Bethesda, MD, USA.
Bio-Protocol
|July 5, 2016
Summary
This study details a protocol for isolating mouse embryo fibroblasts (MEFs) from mouse embryos. These MEFs are crucial for cancer research, enabling analysis of genetic alterations and tumor suppressor gene functions.
Area of Science:
- Cell Biology
- Genetics
- Cancer Research
Background:
- Mouse embryo fibroblasts (MEFs) are vital for studying genetic alterations' cellular effects.
- MEFs facilitate research into overcoming senescence and indefinite proliferation in cancer.
- Immortalized MEFs can acquire mutations leading to tumor formation.
Purpose of the Study:
- To present a protocol for isolating primary MEFs.
- To enable the analysis of the tumor suppressor gene *DLC1* in cellular transformation.
- To provide MEFs suitable for biochemical assays and genetic manipulation.
Main Methods:
- Isolation of MEFs from day 13.5-14.5 mouse embryos.
- Culturing of primary MEFs for experimental use.
- Utilizing genetically engineered mouse strains for MEF derivation.
Main Results:
- A reliable protocol for MEF isolation was established.
- MEFs derived are suitable for downstream applications.
- The protocol supports the study of genes like *DLC1* in transformation.
Conclusions:
- The described protocol effectively yields MEFs for research.
- This method supports the investigation of genetic alterations in cancer development.
- MEFs are a valuable tool for dissecting cellular transformation pathways.
