Somatic hybridisation, mutant selection and direct gene transfer are attractive prospects for crop improvement using protoplast technology provided a protoplast-to-plant regeneration system is available. Such a system, using a feeder layer technique, has been developed for protoplasts isolated from embryogenic cell suspensions of Gossypium hirsutum L. cv. Coker 312 (Peeters et al., 1994), but improvements are needed to increase the plant regeneration frequency.