Description
- Abstract:
- Alzheimer’s disease (AD) is the most common form of dementia, estimated to affect over 6 million individuals in the United States in 20241,2. Despite its prevalence, there is still no substantial treatment or cure for this terminal disease. As a result, significant resources have been invested in expanding AD research. Recent advances in AD research have expanded the use of in vitro tools with the use of human stem cells and 3D models, as well as transgenic animals modified to carry known genetic mutations and risk genes associated with AD. The central aim of this thesis work was to characterize the use of the AD transgenic rat in a 3D primary cortical cell culture model. Using a previously developed 3D primary cortical culture model47, both wild-type (WT) and TgF344-AD transgene positive ‘microtissues’ were created and cultured over four extended periods (DIV 2, 7, 28, 60) and then labeled with an immunofluorescent tag for confocal microscope imaging. The images were quantitatively evaluated using cell counts, mean fluorescence intensity, and 2D Sholl analysis. Throughout the culture, the supernatants of TgF344-AD ‘microtissues’ were collected and examined for amyloid-beta-42 via ELISA and later evaluated using immunofluorescence. We observed significant changes in neural progenitor cell counts and significant changes in the mean fluorescence intensity of GFAP-positive astrocytes across the duration of the culture in wild-type microtissues. Additionally, significant changes in microglia morphology, marked by longer and more ramified extensions, were observed at day in vitro (DIV) 28 compared to DIV 2. Significant changes in secreted Amyloid Beta (42) were detected by ELISA between WT and TgF344-AD microtissues, as well as significant increases between DIV 6 and DIV 28 in TgF344-AD tissues. In conclusion, this 3D culture system exhibits a diverse and dynamic molecular and cellular environment, capable of being cultured for up to 60 days without an additional scaffold or serums, and applicable for the study of the intricate multicellular mechanisms associated with diseases like AD, with integration of the TgF344-AD rat model.
- Notes:
- Thesis (Sc. M.)--Brown University, 2024
Citation
Drexler, Abigail Rose,
"Investigation and quantification of cell-type diversity in three-dimensional primary cortical tissue model of Alzheimer’s Disease"
(2024).
Biomedical Engineering Theses and Dissertations.
Brown Digital Repository. Brown University Library.
https://repository.library.brown.edu/studio/item/bdr:4tcd8dey/
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Biomedical Engineering Theses and Dissertations
Theses and Dissertations for the Biomedical Engineering department....