1. KG1a is a myeloid leukemic cell (10). Cells were grown in a-minimal essential medium supplemented with penicillin (100 U/mL), streptomycine (100 pg/mL), glutamine (2 mmol/L), and 10% fetal calf serum (Gibco, BRL, Invitrogen SARL, Cergy Pontoise, France).
2 Megakaryocytes from CD34+ cells: CD34+ cells were obtained from human bone marrow of healthy patients undergoing hip surgery with their informed consent. Briefly, cells were separated over a Ficoll-metrizoate gradient (Ficoll-separating solution; Biochrom KG, Berlin, Germany); then, CD34+ cells were isolated using the immunomagnetic beads technique (Miltenyi, Biotec, France ). The purity was usually in the range 70 to 90%. CD34+ cells were grown for 5 d in Iscove's modified Dulbeco's medium (Gibco BRL) containing penicillin (100 U/mL), streptomycine (100 pg/mL), glutamine (2 mmol/L; Sigma Chemical Co, St Louis, MO), a-monothioglycerol (76 nmol/L; Sigma), 1.5% deionized bovine serum albumin (BSA; Cohn fraction V; Sigma); 1/100 of insulin-transferrin selenium X (Gibco BRL); and sonicated lipids (20 pg/mL). The culture medium was supplemented with PEG-rhuMGDF (10 ng/mL) and recombinant human stem cell fac tor (SCF 50 ng/mL; generous gifts from Kirin Brewery, Tokyo, Japan, and Amgen, Thousand Oaks, CA, respectively).
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