Date of Award
2024
Document Type
Thesis
Degree Name
Master of Science (MS)
Department
Biological Sciences
Committee Chair
Kyung-Ho Roh
Committee Member
Luis Cruz-Vera
Committee Member
Carrie A. Deans
Research Advisor
Kyung-Ho Roh
Subject(s)
Cultures (Biology), B cells, Germinal centers, Coating, Spheroidal state
Abstract
B cells are critical components of the adaptive immune system that proliferate and differentiate within the secondary lymphoid organs. Traditional two-dimensional (2D) cell cultures fall short of replicating the intricate structures and dynamic evolution of 3D tissue environments, prompting more physiologically pertinent in vitro models. Our approach employs ultra-hydrophilic surfaces engineered from methacrylated N-hexanoyl glycol chitosan to cultivate a three-dimensional (3D) co-culture of CD40L-expressing MS5 stromal cells and naive B cells derived from the peripheral blood mononuclear cells (PBMCs) of healthy human donors. This novel system aims to mirror the structural and cellular intricacies of germinal centers within the secondary lymphoid organs. A comparative analysis was performed to evaluate the effects of 2D and 3D co-culture systems by examining B cell proliferation, differentiation, isotype class switching, and the creation of germinal center (GC) like structures. Our results indicate that the 3D co-culture model markedly improves the physiological accuracy of simulated germinal centers by enabling enhanced isotype class switching and differentiation into effector phenotypes. Notably, the 3D spheroids developed into dynamically evolving spatial organization akin to the dark and light zones found in natural germinal centers. There is tremendous potential to fine-tune the efficacy of this model by adjusting the dosage of CD40L and combinatorial variations of the soluble factors. Thus, this 3D model offers more authentic representations of B cell biology within the secondary lymphoid organs, which is potentially useful for advancing immunology research and the development of new immunotherapeutic strategies.
Recommended Citation
Kumar, Ananta, "3D co-culture of human B lymphocytes with CD40L-expressing fibroblasts using an ultra-hydrophilic surface for in-vitro mimicry of secondary lymphoid organs" (2024). Theses. 727.
https://louis.uah.edu/uah-theses/727