This study was conducted to determine how 3D cultured trophoblasts' secreted factors impact NK-92 activation and cytotoxicity during early pregnancy. In this study, 6 week gestational age human cytotrophoblast stem cells (iCTB) were cultured in 2D, 3D matrigel, and 3D synthetic hydrogels composed of 20 kDa 4-arm poly(ethylene glycol)-maleimide (PEG-Mal) modified with a GFOGR adhesive ligand (1 mM) and crosslinked with dithiothreitol (DTT), a non-degradable crosslinker. On day six, trophoblast supernatants were collected to investigate the influence of trophoblast organoid secreted factors on activated NK cell phenotype, measured by CD107a expression and levels of IFNγ secretion. Here we demonstrate that NK-92 cells possess a dNK2-like phenotype, and that supernatants of cytotrophoblasts cultured in 2D and synthetic hydrogels, but not matrigel, reduce activated NK-92 cytokine secretion.

Pelvic organ prolapse (POP) is a condition involving the weakening of the pelvic floor, with a prevalence of up to 50% of women experiencing the condition to some degree. Individuals with the condition are susceptible to multiple symptoms include vaginal protrusion, dyspareunia, and difficulties with waste excretion. Risk factors are common and numerous for POP, and the economic burden of the condition poses a significant cost to nations worldwide. For many years, the primary solution to POP was the usage of transvaginal meshes, often composed of polypropylene, but rising reports of harmful side effects have led to their recall. Due to this, the space is open for novel solutions, and treatments based in regenerative medicine are on the rise. One such potential treatment is the usage of functionalized polyvinyl alcohol scaffolds to support the regeneration and strengthening of the pelvic floor. To validate the usage of this scaffold, this study focuses on the biocompatibility of the scaffolds, with specific focus on the maintenance of cell viability and proliferation on the scaffold. Through usage of metabolic assays and fluorescence microscopy, scaffolds composed of functional polyvinyl alcohol with cellulose have shown promise in supporting the cell types necessary for reconstructing the pelvic floor.

In order to determine whether the spatial organization of FRCs and their expression of maturation markers (such as Ltbr) are altered with age, I performed immunofluorescence on frozen and cryosectioned whole lymph nodes from young and aged mice. My second aim was to perform RT-qPCR and flow cytometry in order to determine whether FRCs from aged mice have altered expression of maturation markers when compared to young mice. Thus, the goal of the honors thesis research was to determine whether lymph node FRCs in the aged mouse exhibit signs of impaired maturation in their protein and gene expression. As the immune system is profoundly impacted by aging, my project supports a cellular mechanism by which defects in aged tissues disrupt immune cell function. Therefore, understanding the age-associated decline in host defense could provide new avenues for the treatment of many diseases of which the elderly are most vulnerable, in particular re-emerging and novel pathological agents such as COVID-19.