Date of Award

Spring 2026

Abstract

The last 40 years of in vitro cell and tissue biology have relied heavily on tissue culture plastic (TCPS), which offers high transparency and a simple, user-friendly method for growing adherent cells. Unfortunately, TCPS has limitations, such as its rigidity, which may alter cell shape, disrupt cell signaling, and ultimately affect cell fate. The poor control over the adsorption of proteins to TCPS and the fact that surface chemistry is fixed on this material are only two examples of the limitations that motivate the development of hydrogel-based culturing platforms, during which the mechanical properties, charge, hydrophobicity, and ligand presentation can be better controlled to investigate and modulate mammalian cell behavior. To overcome the limitations of the previous study, we developed a library of dextran-based 2D hydrogel substrates by introducing carboxymethyl (anionic) groups and methacrylate groups onto a dextran backbone to form carboxymethyl dextran methacrylate (CMD-MA). This allowed me to modify the hydration of the surface systematically, the hydrophobicity of the interfacial regions, and ultimately the amount of protein adsorption on substrates designed as an alternative to TCPS.In recent years, coupling reactions have gained increasing attention for cross-linking cell-encapsulating hydrogels under biocompatible, chemoselective, and tunable conditions. In the next chapter, we created a β-thiolactone platform derived from penicillamine that addressed the historical limitations of native chemical ligation (NCL). We demonstrated that this approach supports rapid and spontaneous gel formation in fully water-soluble environments, it is biocompatible and eliminates unpleasant odors, as well as N-terminal cysteine is selectively targeted over competing biological thiols. This chemistry allowed us to develop PEG-based hydrogels that can be used for both two-dimensional and three-dimensional cultures, providing a cost-effective, scalable. And biorthogonal alternatives to traditional thio-Michael, strain-promoted azide-alkyne cycloaddition (SPAAC), and inverse electron-demand Diels-Alder (IEDDA) strategies.

Document Type

Dissertation

First Advisor

Nathan J Oldenhuis

Second Advisor

Carmela Amato-Wierda

Third Advisor

Brittany White-Matthieu

Department or Program

Chemistry

Degree Name

Doctor of Philosophy

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