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  • A synthetic human 3D in vitro lymphoid model enhancing B-cell survival and functional differentiation.

A synthetic human 3D in vitro lymphoid model enhancing B-cell survival and functional differentiation.

iScience (2023-01-03)
Maaike V J Braham, Rob S van Binnendijk, Anne-Marie M Buisman, Reina E Mebius, Jelle de Wit, Cécile A C M van Els
ABSTRACT

To investigate B-cell differentiation and maturation occurring in the germinal center (GC) using in vitro culture systems, key factors and interactions of the GC reaction need to be accurately simulated. This study aims at improving in vitro GC simulation using 3D culture techniques. Human B-cells were incorporated into PEG-4MAL hydrogels, to create a synthetic extracellular matrix, supported by CD40L cells, human tonsil-derived lymphoid stromal cells, and cytokines. The differentiation and antibody production of CD19+B-cells was best supported in a 5.0%-PEG-4MAL, 2.0 mM-RGD-peptide composition. The 3D culture significantly increased plasmablast and plasma cell numbers as well as antibody production, with less B-cell death compared to 2D cultures. Class switching of naive CD19+IgD+B-cells toward IgG+ and IgA+B-cells was observed. The formation of large B-cell clusters indicates the formation of GC-like structures. In conclusion, a well-characterized and controllable hydrogel-based human 3D lymphoid model is presented that supports enhanced B-cell survival, proliferation, differentiation, and antibody production.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
IgM from human serum, reagent grade, ~95% (HPLC), buffered aqueous solution
Sigma-Aldrich
DAPI, for nucleic acid staining
Sigma-Aldrich
IgA from human serum, reagent grade, ≥95% (HPLC), buffered aqueous solution