cytokine-driven liquid culture differentiation assays
Protocol for unilineage-restricted differentiation of human CD34 cells
medium: QBSF without serum, two weeks
start with 10.000 cells
Erythroid differentiation:
Epo (3 U/ml), IL-3 (0.01 U/ml), GM-CSF (0.001 ng/ml)
Granulocytic differentiation:
IL-3 (1 U/ml), GM-CSF (0.1 ng/ml), G-CSF (500 U/ml)
Megakaryocytic differentiation:
Tpo (100 ng/ml), IL-3 (0.01 u/ml), KL (10 ng/ml), IL-6 (10 ng/ml)
Monocytic differeniation:
40% FCS, IL-6 (1 ng/ml), FL (100 ng/ml), M-CSF (200 U/ml)
B-cell differentiation:
IL-7 (100 ng/ml), FL (10 ng/ml)
Protocol - liquid culture cytokine-driven differentiation of human CD34 cells
plate 50.000 transduced CD34+ cells in 4 ml Iscove’s modified Dulbecco’s medium (IMDM, Gibco-Life technologies) in the presence of 10% FBS, IL-3 (20 ng/ml), IL-6 (20 ng/ml), c-Kit ligand (KL), Flt-3 ligand (FL) and thrombopoietin (TPO) (20 ng/ml each)
Laatst gewijzigd: | 19 november 2012 16:46 |