Short-term culture for acute myeloid leukemia blast cells

Abstract

This study was carried out to separate acute myeloid leukemia (AML) blast cells and studies their proliferation in short-term culture. The separation procedure include three steps; Ficoll gradient separation, depletion of macrophages and depletion of (lymphocytes and of monocytes) for preparation of highly pure native AML blast cells from blood samples collected from patients with moderate blast percentage. Results showed that this procedure is an inefficient due to a decrease in total cell number and contamination with other cells after each separation step. Proliferation of native AML blast cells in short term-culture by cultivating isolated AML blast cells in RPMI medium supplemented with 20% human plasma at a concentration 1×106/ml in the presence and absence of colony -stimulating factor which was provided by conditioned media (PHA-leucocytes-, plasmacytoma cell line- and Hep-2 conditioned medium. The effect of each conditioned medium on proliferation of AML blast cells was studied separately. Results showed that plasmacytoma cell line conditioned medium didnot stimulate the proliferation of native AML blast cells, while cells seeded on media containing 10%PHA-LCM showed an increase in cell number and growth of the cells was observed for approximately 3 days and then decreased.