Hey everyone, I am looking at the influence of systemic inflammation on myelopoiesis.
Does anyone of you know a suitable model system to differentiate human iPSCs (induced pluripotent stem cells) into granulocytes / neurophils or as far into that direction, as possible?
Thanks and with kind regards!
Dear Willem!
Please You look at the articles:
Article Molecular analysis of neutrophil differentiation from human ...
Differentiation of mature neutrophils from human iPSCs by 4-stage discontinuous culture
Article Neutrophil Differentiation From Human-Induced Pluripotent Stem Cells
(attached)Differentiation of iPS cells Methods used for the initial differentiation of iPS cells and cell sorting were based on earlier reports (Umeda et al., 2004, 2006). Briefly, trypsin-treated undifferentiated iPS cells were transferred onto OP9 cells and cultured with 20 ng/ml vascular endothelial growth factor (VEGF) (R&D Systems). After 10 days, the induced cells were harvested with cell dissociation buffer (Invitrogen), and sorted TRA 1-85þCD34þVEGFR-2high cells were transferred onto fresh OP9 cells in six-well plates at a concentration of 3 103 cells per well. Sorted cells were cultured in a-MEM (Invitrogen) containing 10% FCS (Sigma, St Louis, MO), 50mM 2- mercaptoethanol, 20 ng/ml interleukin (IL)-3, 100 ng/ml stem cell factor (SCF) (R&D Systems), and 10 ng/ml thrombopoietin (TPO) for 20 days. On day 20 after cell sorting, cytokines were changed into 20 ng/ml IL-3 and 10 ng/ml granulocyte colony-stimulating factor (G-CSF). IL-3, TPO and G-CSF were kindly provided by Kyowa Hakko Kirin
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