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Gelatin-Based 3D Microgels for In Vitro T Lineage Cell Generation
journal contribution
posted on 2020-03-11, 20:19 authored by Anisha
B. Suraiya, Michael L. Hun, Vinh X. Truong, John S. Forsythe, Ann P. ChidgeyT cells
are predominantly produced by the thymus and play a significant
role in maintaining our adaptive immune system. Physiological involution
of the thymus occurs gradually with age, compromising naive T cell
output, which can have severe clinical complications. Also, T cells
are utilized as therapeutic agents in cancer immunotherapies. Therefore,
there is an increasing need for strategies aimed at generating naive
T cells. The majority of in vitro T cell generation studies are performed
in two-dimensional (2D) cultures, which ignore the physiological thymic
microenvironment and are not scalable; therefore, we applied a new
three-dimensional (3D) approach. Here, we use a gelatin-based 3D microgel
system for T lineage induction by co-culturing OP9-DL4 cells and mouse
fetal-liver-derived hematopoietic stem cells (HSCs). Flow cytometric
analysis revealed that microgel co-cultures supported T lineage induction
similar to 2D cultures while providing a 3D environment. We also encapsulated
mouse embryonic thymic epithelial cells (TECs) within the microgels
to provide a defined 3D culture platform. The microgel system supported
TEC maintenance and retained their phenotype. Together, these data
show that our microgel system has the capacity for TEC maintenance
and induction of in vitro T lineage differentiation with potential
for scalability.
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HSC2 D culturesT cell generation studiesVitro T Lineage Cell Generation T cellsT lineage differentiationFlow cytometric analysisTEC maintenance3 D environmentgelatin-based 3 D microgel systemT cell outputT lineage induction3 D culture platformmouse fetal-liver-derived hematopoieticthymic epithelial cellsGelatin-Based 3 D Microgelsco-culturing OP 9-DL cellsmicrogel systemT cells
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