Routine maintenance and storage of lepidopteran insect cell lines and baculoviruses

The various methods for maintaining (a.k.a., subculturing, splitting, or passaging) established lepidopteran cell lines are described. Three procedures are presented that are appropriate for different cell lines dependent upon the growth characteristics (in particular, cell attachment properties) of the cells of interest. In addition to the routine maintenance of cells in active culture, methods are also described for both short (low temperature) and long-term (frozen in liquid nitrogen) storage of cell lines, as well as quality control procedures for the cultures. Methods for storing baculoviruses for use in cell cultures and issues of concern when using cell cultures for their production and study are also described.