Maintenance and use of F9 teratocarcinoma cells

F9 cells are an established cell line of mouse embryonal carcinoma cells that differentiate spontaneously at very low frequency. This chapter discusses the maintenance, differentiation, and use of F9 teratocarcinoma cells. F9 cells can be maintained in a conventional cell culture facility. The F9 cell system provides a means to identify mechanisms that control gene expression upon differentiation. Gene transfer experiments are allowing investigators to examine this regulation at the molecular level. To introduce DNA into F9 cells, a transfection protocol that is a modification of the calcium phosphate DNA precipitation technique has been optimized. This procedure can be used to compare the strength of promoter or enhancer elements linked to a reporter gene upon introduction into either F9 stem or differentiated cells. The transfection procedure can also be used to determine the importanceof sequences present in an mRNA which confer transcript stability or translational control.