Easysep human cord blood cd34 positive selection kit

HEK293T (GTX) cells were provided as a kind gift from Kenneth Cornetta (Indiana University School of Medicine, Indianapolis, IN, USA). Cells were maintained in DMEM supplemented with 10% fetal bovine serum (FBS; Omega Scientific, Tarzana, CA, USA), 1% penicillin and streptomycin, and 1× L-glutamine. Human cord blood was provided by the Cleveland Cord Blood Center (Cleveland, OH, USA), under approved institutional protocols and in accordance with the Declaration of Helsinki. Primary human CD34⁺ cells were isolated using the EasySep Human Cord Blood CD34 Positive Selection Kit according to manufacturer’s instructions (STEMCELL Technologies, Vancouver, BC, Canada). Sorted cell purity was checked by flow cytometry, and only cells with >95% CD34⁺ were used in further experiments. Following purification and purity check, cells were frozen in FBS/10% DMSO by liquid nitrogen until later use. THP-1 and KG-1a cells were obtained from the American Type Culture Collection. Raji cells were a kind gift from David Rawlings (Seattle Children’s Research Institute, Seattle, WA, USA). Frozen G-CSF mPB CD34⁺ cells were purchased from the Co-Operative Center for Excellence in Hematology at the Fred Hutchinson Cancer Research Center (Seattle, WA) under approved institutional protocols.