Biomaterials-Mediated blood stem cell Engineering

Abstract

The field of regenerative medicine and bioengineering presents an exciting opportunity to harness the potential of hematopoietic stem cells (HSCs) through stem cell niche and cellular engineering. These approaches aim to recreate the natural environment of the stem cells and modify protein expression in the cells to enhance the differentiation and therapeutic capabilities of HSCs. This makes enhanced HSCs valuable for treating a range of blood-related diseases and genetic conditions. Our first study is focused on developing a hydrogel-based system that provides a serum-free, feeder-free thymic-like niche for HSCs to develop into T cells. We synthesized hydrogel containing thymic-associated components including immobilized Notch

ligand Delta-like-4 (DLL4), hyaluronic acid, vascular cell adhesion molecule 1 (VCAM1) and collagen-derived gelatin. DLL4-Fc immobilized using conjugated protein G resulted in T cell differentiation. Gelatin addition in the engineered 3D system enhanced T cell differentiation, while VCAM1 had no effect on cell development. The second study is focused on using the platform engineered in the first study to develop a system for generation of T cells from induced pluripotent stem cells (iPSCs) derived HSCs. We created the first feeder-free serum-free 3D system for T cell generation from PSCs-derived cells. Finally, in the third study, we explored the use of lipid nanoparticles (LNPs) loaded with mRNA to modify phenotype of megakaryocyte (MK) progenitors derived from HSCs. We found an optimal formulation of LNPs for MK progenitor transfection and engineered the cells to produce coagulation factor VII, FVII, which decreased clot time in the presence of FVII-deficient plasma. Our study highlights the significant potential of biomaterials to engineer HSCs into functional T cells and megakaryocytes, thus opening up new avenues in therapy development.