Date of Award

2026

Document Type

Open Access Master's Report

Degree Name

Master of Science in Biological Sciences (MS)

Administrative Home Department

Department of Biological Sciences

Advisor 1

Smitha Rao Hatti

Advisor 2

Rupali Datta

Committee Member 1

Xiaohu (Mark) Tang

Abstract

Traditional 2D cell culture systems provide a controlled environment for studying cellular mechanisms, but they fail to emulate the complex 3D structure and mechanical stimuli present in native tissue. This study aimed to develop a tunable in vitro environment that more closely mimics the extracellular matrix by integrating porcine gelatin hydrogels with electrospun nanofiber scaffolds to support 3D cell growth. Gelatin crosslinked with microbial transglutaminase (mTG) was synthesized and optimized to create a viscous substrate capable of supporting cell movement and growth, while polycaprolactone nanofibers with aligned, mesh, and honeycomb morphologies were fabricated to introduce structural anisotropy similar to connective tissue. Human breast cancer cells (MCF7-GFP) and mus musculus (mouse) mammary carcinoma cells (4T1) were cultured on gelatin, nanofibers, and combined nanofiber gelatin matrices to evaluate cellular behavior, morphology, and scaffold compatibility. Fluorescence microscopy and time lapse imaging were used to monitor cell viability and growth patterns. The results demonstrate that cells remain viable on nanofiber scaffolds and can integrate with gelatin-fiber matrices, producing observable changes in morphology and movement compared to conventional culture plates. This work establishes a foundational protocol for integrating gelatin and nanofibers into a 3D cellular environment, providing a platform that can be further developed for studies of tumor microenvironments, wound healing, and other complex tissue behaviors in vitro.

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