TY - JOUR
T1 - A synthetic human 3D in vitro lymphoid model enhancing B-cell survival and functional differentiation
AU - Braham, Maaike V. J.
AU - van Binnendijk, Rob S.
AU - Buisman, Anne-Marie M.
AU - Mebius, Reina E.
AU - de Wit, Jelle
AU - van Els, C. cile A. C. M.
N1 - Funding Information: We would like to thank: Mylène de Ruijter (Department of Orthopedics, University Medical Center Utrecht, the Netherlands) for her help measuring the mechanical properties of the PEG-4MAL gels. Reza Nadafi (Department of Molecular Cell Biology and Immunology, Amsterdam University Medical Center, the Netherlands) for the isolation of LSCs. Martien Poelen and Maarten Emmelot for the radiation of CD40LCs. Pieter Dingemanse, Gaby Smits, and Pieter van Gageldonk for their assistance in the set up and usage of the fluorescent bead-based multiplex immunoassay. Nening Nanlohy for her help measuring culture supernatants and the subsequent analysis (all Center for Infectious Disease Control, National Institute for Public Health and the Environment, the Netherlands). The graphical abstract and cartoons shown in Figures 1, 2, 3, and 4 were created with BioRender.com, This research is part of the Target-to-B consortium, a collaboration project that is financed by the PPP Allowance made available by Top Sector Life Sciences & Health to Samenwerkende Gezondheidsfondsen (SGF) in the Netherlands under projectnumber LSHM18055-SGF to stimulate public-private partnerships and co-financing by health foundations that are part of the SGF. We acknowledge the support of patient partners, private partners, and active colleagues of the Target-to-B consortium (see website: www.target-to-b.nl). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the article. M.V.J.B. designed and carried out the experiments and analyzed the data. R.S.V.B. A.M.M.B. R.E.M. J.D.W. and C.A.C.M.V.E. supported data analyzation. M.V.J.B. wrote the article. R.S.V.B. A.M.M.B. R.E.M. J.D.W. and C.A.C.M.V.E. revised the article. J.D.W. and C.A.C.M.V.E. supervised the experiments. The authors declare no competing interests. Publisher Copyright: © 2022 The Authors
PY - 2023/1/20
Y1 - 2023/1/20
N2 - 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.
AB - 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.
KW - Bioengineering
KW - Biological sciences
KW - Cell engineering
KW - Tissue Engineering
UR - http://www.scopus.com/inward/record.url?scp=85144418196&partnerID=8YFLogxK
U2 - https://doi.org/10.1016/j.isci.2022.105741
DO - https://doi.org/10.1016/j.isci.2022.105741
M3 - Article
C2 - 36590159
SN - 2589-0042
VL - 26
JO - iScience
JF - iScience
IS - 1
M1 - 105741
ER -