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Morphological and cytochemical characteristics of cells obtained from Shay chloroleukaemia tumour growing subcutaneously in the rat and after 0.5–16 d of diffusion chamber culturing were assayed using seven different cytochemical procedures. Further, the cytochemical characteristics of the recent subline of Shay chloroma were compared with those reported in 1962 by other workers. The chloroleukaemia cells did undergo some maturation in diffusion chamber cultures, but full differentiation to polymorphonuclear cells was not seen. The most striking change in the cytochemical pattern of chloroleukaemia cells during the diffusion chamber culturing was a significant increase in alkaline phosphatase activity. There were some cytological differences in the recent chloroma tumour compared with that described 10 years ago. The observations made in the present work show that all properties of cells after a neoplastic transformation are not irreversible: many characteristics of malignant chloroleukaemia cells can alter when these cells are maintained in different host animals or in the diffusion chamber milieu.