Derivation of primary mouse embryonic fibroblast (PMEF) cultures.

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.