Self-assembling peptide-based hydrogels can serve as a versatile platform for a variety of biological applications. The wide range of possibilities for modulation of the properties of these hydrogels allow this platform to be broadly applied to a multitude of cell-based assays based on the specific demands of the system. Here, we demonstrate this flexibility of a self-assembling peptide-based hydrogel system by determining the effects of stiffness, charge, and arginylglycylaspartic acid (RGD) content of the hydrogels used in a cell-based culture and how these factors can be manipulated to maximize and promote cell proliferation. NIH 3T3 fibroblast cells were seeded as a cell spheroid and the hydrogel was added to the culture as a free-standing entity. Through this study, an optimum range of stiffnesses and a minimum viable charge were defined for the surrounding hydrogel environment of the fibroblasts for their proliferative growth, while the infusion of RGD in hydrogels allowed for proliferative activity outside of the charge ranges of hydrogels without the sequence. This evaluation of in vitro conditions for a specific cell type and platform can be applied to a broad scope of cell-based assays to study the proliferation and expansion of cells under externally applied experimental conditions. The versatility in the synthesis of an in vitro assay can allow for its use in various ranges of applications that depend on studying the effects of external environments on cell proliferation. This provides an effective means of developing in vitro assays for application in research dependent on a firm understanding of the effect of many external conditions in a cell-based platform, making it possible to extend these principles to promote advances in immune, microbiological, and oncological research and pharmacology.
David Belair; Vicki Workman
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