School of Science Division of Life Science 13 Division of Life Science Molecular Regulation of Skeletal Muscle Stem Cell Quiescence and Activation Supervisor: CHEUNG Tom / LIFS Student: CHENG Man Hoi Adam / BIOT Course: UROP 1100, Spring Both satellite cells and FAP were isolated and cocultured to be tracked by machine learning tools using live cell imaging. In this report, a review on the molecular characteristics of stem cells and the FAP was done. Cell biological techniques such as flow cytometry and live cell imaging were done to achieve the isolation and tracking of the satellite cell and FAP interactions. LAF training is also being done throughout the project for the manipulation and supply of mice. As a practice for the project, C2C12 myoblast immortal cell line was cultured, and live cell imaging was performed over a period of 12 hours to capture the migrations and interactions of C2C12 cells in the culture environment. However, improvements on how to perform the technique can be made in future imaging. Molecular Regulation of Skeletal Muscle Stem Cell Quiescence and Activation Supervisor: CHEUNG Tom / LIFS Student: LEE Wing Yan / BIBU Course: UROP 1100, Spring Satellite cells (SCs) are stem cells that facilitate the growth and repair of skeletal muscles. Understanding the mechanisms in which miRNAs regulate SCs is key to addressing the causes for defects in skeletal muscle regeneration. We are currently studying the role of miR-653 in the regulation of SCs. In this project, the genotype of individuals of the experimental mouse line were verified, and plasmids were prepared for the study of the activity of miR-653 on the gene CDK4. Interaction between miR-653 and CDK4 were tested through a dualluciferase assay. It was found that CDK4 is not a target of miR-653. Molecular Regulation of Skeletal Muscle Stem Cell Quiescence and Activation Supervisor: CHEUNG Tom / LIFS Student: NGAI Man Yee / BCB Course: UROP 2100, Fall UROP 3100, Spring This study employs the Assay for Transposase Accessible Chromatin with high-throughput sequencing (ATACseq) to explore chromatin accessibility across various states of stem cell differentiation, offering insights into mechanisms that facilitate stem cell potency and tissue regeneration. Stem cells, characterized by their ability to differentiate along a potency continuum from totipotent to unipotent states, are pivotal in developmental biology and regenerative medicine. Utilizing the ENCODE ATAC-seq pipeline, data were systematically analyzed to annotate nucleosome-free regions, highlighting the dynamic epigenetic landscape during differentiation (Dong et al., 2022). Troubleshooting various computational challenges, including issues with the SRA Toolkit, Caper, and Conda dependencies, underscored the complexities of bioinformatics workflows. Future work will compare the efficacy of Singularity and Conda environments to optimize pipeline execution.
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