Penicillin streptomycin

4T1 and 4T1-66cl4 cells (kindly provided by Dr. Claudia Chiodoni, Department of Experimental Oncology and Molecular Medicine, Istituto Nazionale dei Tumori, Milano, Italy) were grown in Dulbecco’s modified Eagle medium (DMEM; Lonza) supplemented with 10% fetal bovine serum (FBS; Sigma), 1% penicillin/streptomycin (Lonza), 1% l-glutamine (Lonza), 1% sodium pyruvate (Lonza), and 1% Hepes (Lonza). B16F10, kindly provided by Prof. Massimiliano Mazzone (Vesalius Research Center, Leuven, Belgium), were grown in DMEM (Lonza) supplemented with 10% FBS (Sigma), 1% penicillin/streptomycin (Lonza), and 1% l-glutamine (Lonza). 4T1-luc from PerkinElmer were grown in RPMI 1640 (Lonza), 10% FBS (Sigma), 1% penicillin/streptomycin (Lonza), 1% l-glutamine (Lonza), 1% sodium pyruvate (Lonza), and 5.4 g/l glucose (Sigma). HL-60 were purchased from American Type Culture Collection and grown in Iscove’s modified Dulbecco’s medium (IMDM; Lonza), 20% FBS (Sigma), 1% penicillin/streptomycin (Lonza), 1% l-glutamine (Lonza), and 1% sodium pyruvate (Lonza), and transfected with pEGFP-N1 ACKR2 or mock vector^{52 (link)} by using the Nucleofector Kits for HL-60 (Lonza) according to the manufacturer’s instructions. Green fluorescent protein-positive cells were sorted for mRNA analysis. Cells were tested for mycoplasma and only mycoplasma-free cells were used.

C1R HLA-A2 cells, Jurkat cells, and J.RT3-T3.5 cells were cultured in RPMI medium supplemented with 10% fetal bovine serum (FBS), 1% penicillin/streptomycin, and 1% L-glutamine (Lonza). The generation of C1R HLA-A2 cells was described previously (15 (link)). JE6.1 Jurkat cells, K562 A2 cells, K562 HLA-A2+CD20 cells, Raji HLA-A2 cells, Raji HLA-A2+CTAG1 cells, and U266 cells were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) supplemented with 10% FBS, 1% penicillin/streptomycin, and 1.5% L-glutamine (Lonza). ALL CM cells were cultured in IMDM containing serum-free supplement and 1% penicillin/streptomycin as described previously (48 (link)). UM3 cells were cultured in IMDM supplemented with 20% FBS, 1% penicillin/streptomycin, 1.5% L-glutamine, and 10 ng/ml IL-6 (Lonza). HEK 293T cells were cultured in Dulbecco’s Modified Eagle Medium (DMEM) supplemented with 10% FBS, 1% penicillin/streptomycin, 1% L-glutamine, and 1 mM HEPES (Lonza). Phoenix Ampho cells were cultured in IMDM supplemented with 10% FBS, 1% penicillin/streptomycin, and 1% L-glutamine (Lonza).

SEM cells (DSMZ ACC 546) were cultured in Iscove’s Modified Dulbecco’s Medium (IMDM) (Thermo Fisher Scientific, Waltham, MA, USA), FBS (gibco by Thermo Fisher Scientific Inc., Waltham, MA, USA), and 1% penicillin/streptomycin (Lonza Group AG, Basel, Switzerland). THP-1 cells (DSMZ ACC16) were cultured in Roswell Park Memorial (RPMI) 1620 medium (Thermo Fisher Scientific, Waltham, MA, USA) with 20% FBS (gibco by Thermo Fisher Scientific Inc., Waltham, MA, USA) and 1% penicillin/streptomycin (Lonza Group AG, Basel, Switzerland). MLLr patient peripheral blood mononuclear cells (PBMCs) were isolated from peripheral blood, received from and performed by the University Children’s Hospital Tuebingen. Written consent was obtained from all patients in compliance with the Declaration of Helsinki and approved by our Institutional Review Board (IRB approval 137/2017BO2). Primary patient cells were cultured in Roswell Park Memorial (RPMI) 1620 medium (Thermo Fisher Scientific, Waltham, MA, USA) with 10% FBS (gibco by Thermo Fisher Scientific Inc., Waltham, MA, USA) and 1% penicillin/streptomycin (Lonza Group AG, Basel, Switzerland).