Akshat
Singh
Sponsor: Jiandi Wan, Ph.D. Chemical Engineering Cultivated meat is a rapidly growing field where cells from animals typically slaughtered for meat are cultured to form meat-like products. Current research on bovine muscle stem cells (MuSC) faces challenges in developing muscle fiber maturity and lack of precise native fiber fabrication in vitro. This project aims to establish conditions that reliably promote cell aggregation and myotube differentiation in bovine MuSC cultures by printing them in a core-shell structure. Agarose-matrigel core-shell bioprinting was used to provide a restrictive, structured environment and a nutrient gradient during MuSC culture. This bioprinted structure mimics the spatial conditions cells experience in vivo and provides a nutrient gradient, facilitating both proliferation and differentiation without the need for environmental or media changes during culture. Current results indicate that higher cell concentrations and smaller filament diameters promote MuSC aggregation and differentiation. Filaments with smaller diameters exhibited certain higher mechanical properties. Within large filament diameters, it was also observed that cells tended to aggregate near the edge of the filaments. Multi-channel filaments were also observed to have cross-barrier myogenic fusion during differentiation. This research is still ongoing, and we are currently investigating the ability of separate MuSC filaments to merge together to form larger, complex structures. Expression and Purification of GST-tagged MAGUK Family Proteins Using a Bacterial Expression System
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Akshat Singh
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Membrane-associated guanylate kinases (MAGUKs) are a conserved family of scaffold proteins that organize submembrane protein networks, enabling the precise spatial alignment of pre- and postsynaptic machinery required for synaptic function. Recombinant expression of MAGUK proteins is essential for detailed biochemical and biophysical characterization of their structural organization and protein-protein interactions. Here, we used a bacterial expression system to overexpress glutathione S- transferase (GST)-tagged MAGUK proteins and purified them by affinity chromatography using FPLC. The gene encoding the MAGUK protein was cloned into a GST fusion vector and transformed into E. coli for IPTG-inducible expression. To maximize protein yield, expression conditions, including temperature, IPTG concentration, and post-induction incubation time, were systematically optimized. Following the overexpression, the cells were harvested and lysed. The GST- tagged protein was purified by affinity chromatography using a Glutathione Sepharose-packed column. The purity of the protein was verified by SDS-PAGE, and concentration was measured using UV-vis spectroscopy. Our results demonstrate a high-yield expression and purification of GST-tagged MAGUK proteins using a bacterial expression system, producing proteins suitable for downstream biochemical and biophysical analyses. Effects of Individual Language Experience on Bilingual Infants' Word Recognition in Code- Switched Sentences Anya Singh
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UC Davis / MED: Pharmacology / 2026
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Akshat Singh