Induction of megakaryocytic differentiation in a newly established human leukemic cell line by 12-o-tetradecanoylphorbol-13-acetate.

BACKGROUND

Biological studies on megakaryopoiesis have been hampered by the scarcity of megakaryocytes in normal bone marrow and difficulty in long term culture. Alternatively, leukemic cell lines with megakaryocytic differentiation potential may provide good models to counter these problems.

METHODS

Leukemic cells from a patient with acute megakaryocytic leukemia were put into long-term culture and established into a cell line which was designated as VGH-MK1. The VGH-MK1 cells were challenged with differentiation agents and/or cytokines, and the differentiation of these cells was examined using morphological, immunocytochemical and surface-marker studies.

RESULTS

Morphologically, VGH-MK1 cells had prominent nucleoli and basophilic cytoplasm with some protrusions, but large cells were occasionally seen. Under regular culture condition, the cells had a doubling time of 36-48 hours. The cloned cell line exhibited markers characteristic of megakaryoblasts after differentiation induction. Specifically, when stimulated with 12-o-tetradecanoylphorbol-13-acetate (TPA), cells became larger and had large or multinuclei. They were induced to express platelet glycoproteins GPIb (CD42b), GPIIb/IIIa (CD41), and GPIIIa (CD61) antigens, but not erythroid nor lymphoid markers. Platelet peroxidase (PPO) activity was also induced. Retinoic acid did not exhibit similar differentiation-inducing effects. In contrast, it counteracted the effects induced by TPA.

CONCLUSIONS

An unique human leukemic cell line, VGH-MK1, has been established here. It could be induced to exhibit some characteristics of megakaryocytic lineage, and may be an useful model for the biological studies of megakaryopoiesis.