The effect of cryopreservation on the ability of primary human cancer cells to form colonies in a two-layer agar system was examined. Although considerable variation occurred, concentrations of 5 or 10% dimethylsulfoxide employed with slow freezing rates allowed survival of colonies in the range 20-40% or greater of nonfrozen controls. The methods used in this study do not require elaborate freezing equipment, and can be used for the cryopreservation of a wide variety of types of cancers.