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Thesis
Optimization and Processing of Cardiac and Islet Samples for RNA Sequencing
Master of Science (M.S.), Drexel University
Jul 2023
DOI:
https://doi.org/10.17918/00001795
Abstract
Heart failure and diabetes are respectively the number one and seven leading causes of death in the US. In the heart, adult cardiomyocytes are terminally differentiated cells that are highly specialized to generate the necessary mechanical force and energetic potential needed to pump blood to the body. Similarly, pancreatic islets are highly specialized cells involved in glucose-mediated insulin and glucagon peptide responses in response to blood glucose levels. Due to the highly specific nature of these cell types, it is not surprising that gene expression patterns are uniquely fine-tuned to accommodate their biological role in physiological and pathological conditions. Recently, research has focused on deciphering metabolic pathways in these cells that are exclusive and altered in various disease states. A significant hurdle in utilizing tissue and/or primary myocytes and islets is the difficulty in handling and obtaining metabolically active and healthy specimens. Here, we aimed at optimizing and processing hearts and islets to obtain high-quality samples suitable for single cell/nuclei RNA sequencing studies. Through the implementation of rigorous quality control measures and the optimization of various parameters, a protocol has been developed for the heart and another for pancreatic islets that allows us to further assess cell-type specific signaling networks by single cell/nuclei RNA-sequencing strategies. The goal of employing these protocols is to explore novel tissue-specific pathways that undergo alterations due to genetic or pharmacological interventions. By unraveling the intricate molecular mechanisms underlying these conditions, new avenues for therapeutic interventions may be discovered, leading to improved patient outcomes and the potential to mitigate the devastating impact of heart failure and diabetes on public health.
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Details
- Title
- Optimization and Processing of Cardiac and Islet Samples for RNA Sequencing
- Creators
- Rohail Bhatti
- Contributors
- Paul McGonigle (Advisor)
- Awarding Institution
- Drexel University
- Degree Awarded
- Master of Science (M.S.)
- Publisher
- Drexel University; Philadelphia, Pennsylvania
- Number of pages
- viii, 31 pages
- Resource Type
- Thesis
- Language
- English
- Academic Unit
- College of Medicine; Pharmacology and Physiology; Drexel University
- Other Identifier
- 991021227814004721