Establishment and characterization of an arsenic‐sensitive monoblastic leukaemia cell line (SigM5)

Abstract

Few human monoblastic cell lines have been characterized to date. We have established the SigM5 cell line from a patient with acute monoblastic leukaemia (FAB M5a). Original leukaemic cells had a karyotype of 47,XY,+8, whereas the cell line showed a stemline clone of 81,XX,Y,Y,1,4,6,7,+8,+8,9,10,10,11,13,16,19[cp], with a minor sideline also present. Cytochemical staining was strongly positive with α‐naphthylbutyrate acetate esterase, particulate positive with Sudan black and weakly positive for myeloperoxidase. Cells were positive for CD13, CD15, CD18, CD23, CD33, CD38, CD45, CD68 and myeloperoxidase. CD14 expression was 3–15%. SigM5 constitutively secreted interleukin (IL)‐2, IL‐8, IL‐10, tumour necrosis factor (TNF)‐α, ferritin, lysozyme, N‐elastase and neopterin upon stimulation with interferon (IFN)‐γ. Cells expressed the proinflammatory mediator macrophage migration inhibitory factor (MIF). All NADPH oxidase subunits were constitutively present, but nitroblue tetrazolium reduction was only detectable upon activation with IFN‐γ. SigM5 monoblasts were sensitive to arsenic trioxide (As2O3) previously not described to induce apoptosis in monoblastic cells. Differing considerably in morphology, immunophenotype and sensitivity to arsenics from the widely used cell lines U937, HL‐60 and THP‐1, SigM5 is a new monoblastic cell line useful for studying leukaemogenesis, monocyte differentiation and tumour cell susceptibility to arsenic compounds.

Functional and Selective Targeting of Adenovirus to High-Affinity Fcγ Receptor I-Positive Cells by Using a Bispecific Hybrid Adapter

“…THP-1 cells were cultured in Iscove modified Dulbecco medium (IMDM) plus 10% FCS, whereas SigM5 cells were propagated in IMDM plus 20% human serum. All cell lines were originally purchased from the American Type Culture Collection, with exception of Kasumi-1 (Department of Human and Animal Cell Cultures, Deutsche Sammlung von Mikroorganismen und Zellkulturen, Braunschweig, Germany) and SigM5 (57). Primary patient AML cells were isolated using the standard therapeutic leukapheresis procedure after obtaining informed consent, typed, and frozen in IMDM plus 50% pooled human serum.…”

Dual effects of arsenic trioxide (As2O3) on non‐acute promyelocytic leukaemia myeloid cell lines: induction of apoptosis and inhibition of proliferation

“…et al, 1999). Moreover, the potency of As 2 O 3 to induce apoptosis was shown for non-APL leukaemia and lymphoma cell lines (Konig et al, 1997;Wang et al, 1998;Lu et al, 1999;Zhu et al, 1999;Puccetti et al, 2000;Walter et al, 2000) as well as other tumour cell lines (Akao et al, 1999;Huang, C. et al, 1999;Zhang et al, 1999;Deng et al, 2000;Chen, F. et al, 2001;Jiang et al, 2001). The necessity of the PML-RARa fusion protein present in APL-cell lines for As 2 O 3 -induced apoptosis thus became questionable.…”

Hematopoietic Expression of Oncogenic BRAF Promotes Aberrant Growth of Monocyte-Lineage Cells Resistant to PLX4720

“…The SigM5 cell line (28) was purchased from DSMZ (German Collection of Microorganisms and Cell Cultures) and maintained in IMDM supplemented with 20% heat-inactivated FCS with splitting saturated cultures at 1:2 every 3-4 days. The heterozygous BRAF T1799A mutation in this cell line was confirmed by sequencing exon 15.…”