A Functional Assay to Assess Toxicity During Murine B Cell Development In Vitro.

Autor: Guilbert C; Lady Davis Institute for Medical Research, Montréal, Québec, Canada., Chou H; Lady Davis Institute for Medical Research, Montréal, Québec, Canada.; Division of Experimental Medicine, McGill University, Montréal, Québec, Canada., Bolt AM; Department of Pharmaceutical Sciences, College of Pharmacy, University of New Mexico, Albuquerque, New Mexico., Wu TH; Lady Davis Institute for Medical Research, Montréal, Québec, Canada.; Division of Experimental Medicine, McGill University, Montréal, Québec, Canada., Luo VM; Lady Davis Institute for Medical Research, Montréal, Québec, Canada.; Department of Microbiology and Immunology, McGill University, Montréal, Québec, Canada., Orthwein A; Lady Davis Institute for Medical Research, Montréal, Québec, Canada.; Division of Experimental Medicine, McGill University, Montréal, Québec, Canada.; Department of Microbiology and Immunology, McGill University, Montréal, Québec, Canada.; Gerald Bronfman Department of Oncology, McGill University, Montréal, Québec, Canada., Mann KK; Lady Davis Institute for Medical Research, Montréal, Québec, Canada.; Division of Experimental Medicine, McGill University, Montréal, Québec, Canada.; Gerald Bronfman Department of Oncology, McGill University, Montréal, Québec, Canada.
Jazyk: angličtina
Zdroj: Current protocols in toxicology [Curr Protoc Toxicol] 2020 Mar; Vol. 83 (1), pp. e91.
DOI: 10.1002/cptx.91
Abstrakt: B lymphocytes, or B cells, are important players in immunity that produce antigen-specific immunoglobulins. As a result, they are involved in various immune-linked pathologies. To better understand, prevent, or treat B cell-associated disease and immunotoxicity, we developed an in vitro assay to model early murine B cell differentiation within the bone marrow. This model uses sorted B cell precursors cultured on a supporting stromal cell layer, which over time acquire markers of further differentiated B cells, such as surface antigens and rearranged immunoglobulin light chain. Importantly, we utilized our in vitro model to validate our previous observations that xenobiotics, such as tungsten and organotins, alter B cell development in vivo. Furthermore, gene expression can be modulated in this model using retroviral transduction, making it amenable to investigating signaling pathways involved in disruption of B cell differentiation. © 2019 by John Wiley & Sons, Inc. Basic Protocol: Assessment of early B lymphocyte differentiation in vitro Support Protocol: Isolation of murine bone marrow Alternate Protocol 1: Addition of recombinant interleukin-7 Alternate Protocol 2: Genetic manipulation via retroviral transduction.
(© 2019 John Wiley & Sons, Inc.)
Databáze: MEDLINE
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