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Megakaryocyte (MK) colony formation has been studied in the chronic phase and in the blast crisis of chronic myeloid leukaemia (CML). Blood cells were grown in plasma clot for 13 d. MKs were subsequently identified by immunofluorescent techniques using two monoclonal antiplatelet antibodies (AN51 and J15). The maturation process was studied by ultrastructural methods. A marked increase in the number of circulating CFU-MK was observed in all the 10 cases studied prior to chemotherapy (70-fold increase per ml of blood). No significant modification in the regulation of MK colony formation as compared to that of normal subjects was observed. The predominant abnormality in maturation in culture was the occurrence of many hypoploid MKs (microMKs). However, the cytoplasmic maturation of the MKs was identical to that of normal subjects with occasional platelet shedding. Since microMKs predominated in some patients, scoring of MK colonies in CML necessitated immunofluorescent labelling to permit identification of MKs. During the blast crisis, MK colony formation occurred in four out of five patients with an extremely high plating efficiency in the case of promegakaryoblastic transformation. In contrast, MK colonies could not be grown from blood samples of patients with acute leukaemia, including two cases of promegakaryoblastic leukaemia. Maturation of MKs in blast crisis was identical to that of the chronic phase. Furthermore, after short periods of culture in liquid medium, circulating promegakaryoblasts from patients in blast crisis matured with the consequent production of alpha-granules and demarcation membranes. These results confirm the contention that CML represents a pluripotent stem-cell disease, involving the MK lineage, and suggest that the block in maturation during the acute phase can be overcome in vitro.

Type

Journal article

Journal

Br J Haematol

Publication Date

05/1982

Volume

51

Pages

131 - 146

Keywords

Cell Differentiation, Cells, Cultured, Colony-Forming Units Assay, Fluorescent Antibody Technique, Hematopoietic Stem Cells, Humans, Leukemia, Myeloid, Megakaryocytes, Microscopy, Electron