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Updated: Jul 7, 2026
Preparation of Mouse Embryonic Fibroblast Cells Suitable for Culturing Human Embryonic and Induced Pluripotent Stem Cells
Published on: June 21, 2012
Mouse embryo fibroblast (MEF) feeder cell preparation
1Harvard Medical School, Boston, Massachusetts, USA.
Abstract:
Mitotically inactive mouse embryo fibroblasts (MEFs) are commonly used as feeder layers to prevent the differentiation of mouse embryonic stem (ES) cells. This unit describes the isolation of MEFs and the use of g-irradiation or mitomycin C to produce inactivated feeders suitable for the culture of ES cells.
Insights
Mouse embryo fibroblasts (MEFs) are inactivated to serve as feeder layers for culturing mouse embryonic stem (ES) cells. This method ensures ES cells do not differentiate, maintaining their pluripotency for research applications.
Area of Science:
- Cell Biology
- Developmental Biology
- Stem Cell Research
Background:
- Mouse embryonic stem (ES) cells require specific culture conditions to maintain pluripotency.
- Mitotically inactive mouse embryo fibroblasts (MEFs) are essential feeder layers for preventing ES cell differentiation.
Purpose of the Study:
- To describe the isolation of MEFs.
- To detail methods for inactivating MEFs for ES cell culture.
Main Methods:
- Isolation of primary mouse embryo fibroblasts (MEFs).
- Inactivation of MEFs using gamma irradiation.
- Inactivation of MEFs using mitomycin C.
Main Results:
- Established protocols for MEF isolation.
- Demonstrated effective MEF inactivation using two distinct methods.
- Confirmed suitability of inactivated MEFs as feeder layers for ES cell culture.
Conclusions:
- MEFs can be effectively isolated and inactivated for use as feeder layers.
- Gamma irradiation and mitomycin C are reliable methods for MEF inactivation.
- This protocol supports the long-term culture of undifferentiated mouse ES cells.
