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

A Functional Assay to Assess Toxicity During Murine B Cell Development In Vitro.
Curr Protoc Toxicol. 2020 Mar;83(1):e91
Authors: Guilbert C, Chou H, Bolt AM, Wu TH, Luo VM, Orthwein A, Mann KK
Abstract
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.
PMID...

Source: Current Protocols in Toxicology - December 19, 2019 Category: Toxicology Tags: Curr Protoc Toxicol Source Type: research