Cd38 pecy7 hb7

Primary human CD34-enriched cord blood was transduced with lentivirus (pLVX EF1α-IRES-zsGreen; Clontech) overnight, and sorted for GFP expression using a FACSAria II (Becton-Dickinson). For progenitor media assay, cells were, washed and incubated in HPGM medium (Lonza) supplemented with FLT3L, thrombopoietin and stem cell factor (SCF). The cytokines were all purchased from Peprotech and used at 20 ng/ml. 6 days post-culture, differentiation of GFP-positive cells was assessed by flow cytometry using anti-human CD34-APC (8G12) and CD38-PE-Cy7 (HB7) (BD Biosciences). For myeloid differentiation assay, cells were cultured in Myelocult H5100 (Stem Cell Technologies) with 20-µg/mL each of IL-3, SCF, FLT3L, and GM-CSF (Peprotech) for 6 days. Myeloid differentiation was assessed by flow cytometry using anti-human CD33-PE (WM53) and CD14-APC-Cy7 (MφP9) (BD Biosciences). For erythroid differentiation assay, cells were cultured in Stemspan II with erythroid expansion kit (Stemcell Technologies) for 6 days. Erythroid differentiation was assessed by flow cytometry using anti-human GPA-PE (CD235a) (HIR2) and CD71-PE-Cy7 (OKT9) (BD Biosciences).