Honors Theses and Capstones
Date of Award
Spring 2025
Project Type
Senior Honors Thesis
College or School
COLSA
Department
Department of Biological Science
Program or Major
Biology
Degree Name
Bachelor of Science
First Advisor
Nathan Oldenhuis
Second Advisor
Liquing Li
Abstract
The demand for 3D cell cultures has led to an increase in cross-linked hydrogel methodologies. These systems mimic in vivo cellular processes better than their 2D analogs. However, researchers have struggled to develop nontoxic, inexpensive, and kinetically favorable hydrogels for cell encapsulation. Here we present our approach to hydrogel formation using native chemical ligation (NCL). Our use of ß-thiolactone enables hydrogel formation in less than two minutes, significantly faster than previous NCL-based systems that form at 10-20 minutes. Moreover, our NCL system eliminates the cytotoxic free thiols associated with previous work by sequestering the thiol within the PEG backbone. To accomplish this, we functionalized of tetra-armed PEG (Mw = 10,000) with ß-thiolactones and N-terminal cysteines. When dissolved in buffer, the two polymers combine, forming a network with favorable kinetics at 5 wt%. Additionally, our method also allows for post-gelation of attachment motifs and modifications. We have demonstrated the cell viability and adhesion properties of our gel through laserscanning confocal microscopy. We are currently optimizing our NCL system for 3D cell cultures and fiber-pulling platforms.
Recommended Citation
Sager, Dylan M. and Currier, Matthew E., "Hydrogel synthesis via ß-thiolactone mediated native chemical ligation for cell encapsulation: imaging-based assessment of cell viability and spreading" (2025). Honors Theses and Capstones. 890.
https://scholars.unh.edu/honors/890
Included in
Biochemistry Commons, Biomaterials Commons, Organic Chemistry Commons, Polymer Chemistry Commons