Cell Isolation CD34+ cells

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Protocol tips
CD34+ cells were isolated from CB or BM using the CD34 microbead kit (Miltenyi Biotec). Purity of isolated cells was monitored by flow cytometry.
Protocol tips
CD34+ cells were isolated using EasySep™ human cord blood CD34 positive selection kit II (cat# 17896, Stemcell Technologies, Vancouver, BC, Canada) and stored at −150 °C.
Protocol tips
CD34+ HPs of the THC2 patient were isolated from bone marrowaspirate with monocyte-separating medium (TBD) followed by anti- CD34+ magnetic microbead positive selection using EasySep™ Human CD34 Positive Selection Kit II (STEMCELL).
Protocol tips
CD34+ cells were kept for 2-3 days in StemAlpha A medium (StemAlpha) in the presence of Flt3-ligand (50ng/ml), thrombopoietin (50ng/ml) and stem cell factor (50ng/ml).
Protocol tips
CD34+ cells were purified from fresh peripheral-blood mononuclear cells using a complete kit for Human whole-blood CD34+ cells and cultured in StemCell specific medium supplemented with StemSpan CC110 Cytokine cocktail from StemCell (100x).
Protocol tips
The CD34+ fraction was isolated by magnetic bead method as per manufacturer's instructions
Downstream tips
Isolated CD34+ cells were expanded for 3 days in StemPro-34 SFM (Invitrogen, Grand Island, NY, USA) serum free media with SCF, IL-6, TPO and Flt3L (25 ng/ml each, Peprotech, Princeton, NJ, USA).
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