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Updated: Jun 15, 2026
Preparation of Mouse Embryonic Fibroblast Cells Suitable for Culturing Human Embryonic and Induced Pluripotent Stem Cells
Published on: June 21, 2012
Derivation of primary mouse embryonic fibroblast (PMEF) cultures
1Department of Pharmacology, University of Cambridge, Cambridge CB2 1PD, UK.
Abstract:
Primary mouse embryonic fibroblasts (PMEFs) have a number of properties that make them an attractive cell culture model. Relative to other primary explant cultures they are easy to establish and maintain, proliferate rapidly and, as a result, large numbers of cells can be produced from a single embryo within several days following explantation. This allows, for instance, for ready comparison of wild-type and knockout cells derived from the same litter of animals. PMEFs can be expanded through several passages before they reach crisis and can be used to establish cell lines following spontaneous transformation or following derivation from strains carrying mutations, such as in the gene encoding the tumour suppressor Trp53. They have been widely used as feeders to support other cultured cell types, notably embryonic stem cells, as well as for the study of a diverse range of cellular phenomena using microscopic, biochemical and molecular biological techniques. Here, we describe a simple and reliable method for the derivation and maintenance of PMEFs.
Insights
Primary mouse embryonic fibroblasts (PMEFs) offer a robust cell culture model due to their rapid proliferation and ease of maintenance. This study details a simple, reliable method for deriving and maintaining PMEFs for research applications.
Area of Science:
- Cell Biology
- Developmental Biology
- Genetics
Background:
- Primary mouse embryonic fibroblasts (PMEFs) are valuable for cell culture research.
- They are easy to establish, maintain, and proliferate rapidly.
- PMEFs serve as feeder cells for stem cells and are used in various cellular studies.
Purpose of the Study:
- To describe a simple and reliable method for deriving and maintaining PMEFs.
- To highlight the utility of PMEFs in comparative studies (e.g., wild-type vs. knockout).
- To facilitate the use of PMEFs in studying cellular phenomena and establishing cell lines.
Main Methods:
- Derivation of PMEFs from mouse embryos.
- Maintenance and expansion of PMEFs through multiple passages.
- Utilizing PMEFs for feeder layers and cellular research.
Main Results:
- A straightforward protocol for PMEF isolation and culture was established.
- PMEFs can be generated in large quantities from single embryos.
- The method supports the use of PMEFs for various experimental setups.
Conclusions:
- The described method provides a reliable approach for PMEF culture.
- PMEFs are a versatile tool in cell biology and stem cell research.
- This protocol enhances the accessibility and application of PMEFs in scientific investigations.
