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Research Internal Medicine Experimental Hematology
University Medical Center Groningen

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