title: The Culture Repopulation Ability (CRA) Assay and Incubation in Low Oxygen to Test Antileukemic Drugs on Imatinib-Resistant CML Stem-Like Cells.
abstract: Chronic myeloid leukemia (CML) is a stem cell-driven disorder caused by the BCR/Abl oncoprotein, a constitutively active tyrosine kinase (TK). Chronic-phase CML patients are treated with impressive efficacy with TK inhibitors (TKi) such as imatinib mesylate (IM). However, rather than definitively curing CML, TKi induces a state of minimal residual disease, due to the persistence of leukemia stem cells (LSC) which are insensitive to this class of drugs. LSC persistence may be due to different reasons, including the suppression of BCR/Abl oncoprotein. It has been shown that this suppression follows incubation in low oxygen under appropriate culture conditions and incubation times.Here we describe the culture repopulation ability (CRA) assay, a non-clonogenic assay capable - together with incubation in low oxygen - to reveal in vitro stem cells endowed with marrow repopulation ability (MRA) in vivo. The CRA assay can be used, before moving to animal tests, as a simple and reliable method for the prescreening of drugs potentially active on CML and other leukemias with respect to their activity on the more immature leukemia cell subsets.
authors: Department of Experimental and Clinical Biomedical Sciences "Mario Serio", Section of Experimental and Clinical Oncology, Università degli Studi di Firenze, viale GB Morgagni, 50-50134, Florence, Italy. giulia.cheloni@unifi.it.
authors: Giulia
authors: G
authors: Cheloni
authors: Department of Experimental and Clinical Biomedical Sciences "Mario Serio", Section of Clinical Physiopathology, viale G. Pieraccini, 6-50139, Florence, Italy. michele.tanturli@unifi.it.
authors: Michele
authors: M
authors: Tanturli
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology; D000069286:Bortezomib / Q000494:pharmacology; D018929:Cell Culture Techniques / Q000379:methods; D019008:Drug Resistance, Neoplasm / Q000187:drug effects*; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D006801:Humans; D000068877:Imatinib Mesylate / Q000494:pharmacology; D020014:K562 Cells; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000188:drug therapy*; D018942:Macrolides / Q000494:pharmacology; D014411:Neoplastic Stem Cells / Q000166:cytology / Q000187:drug effects*; D010100:Oxygen / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D018942:Macrolides; C571877:salarin C; D000069286:Bortezomib; D000068877:Imatinib Mesylate; D010100:Oxygen
keywords: Hypoxia; Imatinib mesylate; Incubation in low oxygen; Leukemia stem cells; Minimal residual disease