Monomac 1

AML cell lines HL-60 (ACC-3), and MonoMac-1 (ACC-252) were obtained from DSMZ. MDS-L^{34 (link),35 (link)} was kindly provided by Dr. Starczynowski (Cincinnati Children’s Hospital Medical Centre, OH, USA). AML cell lines were cultured in RPMI medium (Biowest) supplemented with fetal bovine serum (FBS, Lonza), 2 mM L-Glutamine (Lonza) and/or 0.1 mM non-essential amino acids (Lonza) according to manufacturers’ recommendations. Primary MDS and CMML blasts were cultured in IMDM (Biowest) supplemented with 3% heat-inactivated FBS, 2 mM L-Glutamine, 20% BIT 9500 Serum Substitute (StemCell Technologies), 5 ng/ml IL3 (Peprotech), 1 mM sodium pyruvate and 5 × 10⁻⁵ M β-mercaptoethanol (Sigma-Aldrich,) and 0.1 mM non-essential amino acids (Lonza).

AML cell lines HL-60 (ACC-3)[30 (link)], KG-1 (ACC-14)[31 (link)], MonoMac-1 (ACC-252)[32 (link)] and Kasumi-1 (ACC-220)[33 (link)] were obtained from DSMZ (Deutsche Sammlung von Mikroorganismen und Zellkulturen) and the human stroma cell line HS-5 was purchased from ATCC (American Type Culture Collection). Experiments were performed within 6 months after receipt or resuscitation. AML cell lines were cultured in complete RPMI medium (PAA laboratories) supplemented with fetal bovine serum (Lonza), sodium pyruvate (Lonza) and/or non-essential amino acids (Lonza) according to manufacturers' recommendations. HS-5 cell line was cultured in complete DMEM medium (PAA laboratories) supplemented with 10% fetal bovine serum (Lonza). Primary AML blasts were cultured in IMDM (PAA laboratories) supplemented with 3% heat-inactivated fetal bovine serum (Lonza), 1x BIT (StemCell Technologies), 5 ng/ml IL3 (Peprotech), sodium pyruvate (Lonza) and β-mercaptoethanol (Sigma).

Human peripheral blood was collected into heparin tubes (Greiner Bio-One, Monroe, NC) from healthy adult volunteers who gave written consent and with specific approval from the Texas A&M University human subjects Institutional Review Board. Peripheral blood mononuclear cells (PBMC) were isolated from the blood using Ficoll-Paque Plus (GE Healthcare Biosciences, Piscataway, NJ), as described previously [4 (link), 29 (link)]. HL-60, K562, THP-1, U937 (ATCC, Manassas, VA), Mono Mac 1 and Mono Mac 6 (DSMZ, Braunschweig, Germany) were grown in RPMI 1640 with 10% bovine calf serum (BCS) (VWR-Seradigm, Radnor, PA) containing 2 mM glutamine, 100 U/ml penicillin, and 100 μg/ml streptomycin (all from Lonza, Walkersville, MD). K562 is a chronic myeloid leukemia cell line [30 (link)], U937 is a lymphoma cell line [31 (link)], and HL-60 [32 (link)], THP-1 [33 (link)], Mono Mac 1, and Mono Mac 6 [34 (link)], are acute myeloid leukemia cell lines. Each individual experiment was performed in duplicate or triplicate, and repeated at least three separate times. For experiments involving human peripheral blood, at least three different donors were used for each experiment. Data are presented as the mean and standard error of the mean (SEM).