Confocal Benchmarking of Human Mesenchymal Stromal Cells in 3-Dimensional Hydrogels
Dr. Sudhir Khetan, Aaron Carretero Benites
Abstract
Mesenchymal stromal cells (MSCs) have emerged as promising therapeutics for immune diseases due to their modulation of immune cells. Their therapeutic effects are mediated through the secretion of immunomodulatory factors packaged in extracellular vesicles (EVs). Control of MSC growth and EV production are critical components of EV therapeutic development, but there is limited understanding of how different aspects of the manufacturing process impact EV reproducibility. This study investigates the influence of the 3D hydrogel microenvironment on MSC-EV immunomodulatory function. Hydrogel design-dependent cell count and morphology are monitored over time using in vivo Calcein staining and confocal microscopy. Preliminary results indicate that MSCs encapsulated in hydrogels that allow encapsulated cell degradation and spreading appear to support better production than either cells in non-degradable hydrogels or in 2D environments. Currently, efforts are focused on optimizing the hydrogel formulation for optimal immunomodulatory factors production.