We report the isolation and regeneration of protoplasts from an embryogenic banana (Musa spp.) cell suspension culture initiated from in vitro proliferating meristems. A high yielding isolation method (up to 6x10(7) protoplasts.ml-1 packed cells) is discussed. Optimal regeneration, with more than 50 % of the protoplasts showing initial cell division, occurred when high inoculation densities (10(6) protoplasts.ml-1) or nurse cultures were applied. Under these conditions, the frequency of microcolony formation was 20-40 %. These microcolonies developed directly, without an intervening callus phase, into somatic embryos which later germinated and formed plantlets.