Graduation Date
Spring 2020
Document Type
Thesis
Program
Master of Science degree with a major in Biology
Committee Chair Name
John W. Steele
Committee Chair Affiliation
HSU Faculty or Staff
Second Committee Member Name
Amy Sprowles
Second Committee Member Affiliation
HSU Faculty or Staff
Third Committee Member Name
Brigitte Blackman
Third Committee Member Affiliation
HSU Faculty or Staff
Fourth Committee Member Name
John Reiss
Fourth Committee Member Affiliation
HSU Faculty or Staff
Subject Categories
Biology
Abstract
Human induced pluripotent stem cells offer a model for human brain development and disease by differentiation into brain organoids; however, current neural culture systems lack the microenvironment, neuronal circuits and connectivity, vascular circulation, and immune system that exist in vivo. After differentiation and development of neuronal and non-neuronal cell types within two formats of cell cultures, we can visualize and recapitulate in vivo protein accumulation, gene expression, and degradative processes such as autophagy. Using RNA extraction, purification methods and reverse transcription I compared traditional monolayer cultures and novel 3-D neural sphere cultures via gene expression analysis. This analysis indicated variable gene expression between formats therefore only monolayer cultures were analyzed for tau protein accumulation with pharmacologic treatments and measured by Western blot. I also report on cell type specific gene expression by transient transfection of plasmid cassette tools and fluorescent microscopy. Here, cell type specific gene targeting is demonstrated in successfully transfected cells. Further development of the tools utilized in this study will significantly expand the field of neurodegenerative research, by giving us the ability to target specific cell types within mixed cultures, and allowing for a more accurate depiction of pathogenesis within diseased cell types.
Citation Style
CBE
Recommended Citation
Anthoney, Kyle H., "Neurodegenerative modeling: tau protein, degradative pathways, and gene expression profiling of human IPSC-derived neural precursors and differentiated 3-D neural sphere versus 2-D monolayer cultures" (2020). Cal Poly Humboldt theses and projects. 384.
https://digitalcommons.humboldt.edu/etd/384
Primer sequences and additional information
Included in
Biology Commons, Biotechnology Commons, Cell and Developmental Biology Commons, Cellular and Molecular Physiology Commons, Medicine and Health Sciences Commons, Microbiology Commons, Molecular Genetics Commons, Neuroscience and Neurobiology Commons