title: Mimicking the Acute Myeloid Leukemia Niche for Molecular Study and Drug Screening.
abstract: Bone marrow niche is a major contributing factor in leukemia development and drug resistance in acute myeloid leukemia (AML) patients. Although mimicking leukemic bone marrow niche relies on two-dimensional (2D) culture conditions, it cannot recapitulate complex bone marrow structure that causes introduction of different three-dimensional (3D) scaffolds. Simultaneously, microfluidic platform by perfusing medium culture mimic interstitial fluid flow, along with 3D scaffold would help for mimicking bone marrow microenvironment. In this study TF-1 cells were cocultured with bone marrow mesenchymal stem cells (BM-MSCs) in 2D and 3D microfluidic devices. Phenotype maintenance during cell culture and proliferation rate was assayed and confirmed by cell cycle analysis. Morphology of cells in 2D and 3D culture conditions was demonstrated by scanning electron microscopy. After these experiments, drug screening was performed by applying azacitidine and cytarabine and cytotoxicity assay and quantitative reverse transcription-polymerase chain reaction (qRT-PCR) for B cell lymphoma 2 (BCL2) were done to compare drug resistance in 2D and 3D culture conditions. Our result shows leukemic cells in 3D microfluidic device retaining their phenotype and proliferation rate was significantly higher in 3D culture condition in comparison to 2D culture condition (p < 0.05), which was confirmed by cell cycle analysis. Cytotoxicity assay also illustrated drug resistance in 3D culture condition and qRT-PCR demonstrated higher BCL2 expression in 3D microfluidic device in contrast to 2D microfluidic device (p < 0.05). On balance, mimicking bone marrow niche would help the target therapy and specify the role of niche in development of leukemia in AML patients.
authors.affiliation: 1 Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine , Tehran, Iran .
authors.forename: Mohammad
authors.initials: M
authors.lastname: Houshmand
authors.affiliation: 2 Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University , Tehran, Iran .
authors.forename: Masoud
authors.initials: M
authors.lastname: Soleimani
authors.affiliation: 3 Stem cell and Tissue Engineering Research Center, Shahroud University of Medical Sciences , Shahroud, Iran .
authors.forename: Amir
authors.initials: A
authors.lastname: Atashi
authors.affiliation: 4 Department of Clinical and Biological Sciences, "S. Luigi Gonzaga" Hospital, University of Turin , Orbassano, Italy .
authors.forename: Giuseppe
authors.initials: G
authors.lastname: Saglio
authors.affiliation: 2 Department of Hematology, Faculty of Medical Sciences, Tarbiat Modares University , Tehran, Iran .
authors.forename: Mohammad
authors.initials: M
authors.lastname: Abdollahi
authors.affiliation: 1 Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine , Tehran, Iran .
authors.forename: Mahin
authors.initials: M
authors.lastname: Nikougoftar Zarif
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D032701:Biomimetics; D001854:Bone Marrow Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D002453:Cell Cycle / Q000187:drug effects; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D018920:Coculture Techniques; D004353:Drug Evaluation, Preclinical; D019008:Drug Resistance, Neoplasm; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy* / Q000378:metabolism / Q000473:pathology*; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D054457:Tissue Scaffolds; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents
keywords: CFSE; bone marrow niche; drug resistance; leukemia; mesenchymal stem cells; microfluidics