Thesis
Optimization of human organotypic brain slice cultures for use as a novel model of neuroHIV
Master of Science (M.S.), Drexel University
Dec 2023
DOI:
https://doi.org/10.17918/00001931
Abstract
Current systems to study neuroHIV often lack the full complexity of the adult brain or suffer in translation to human disease. We aim to overcome these pitfalls by developing an organotypic human brain slice culture system with healthy tissue donated from the incision path of brain surgeries. We hypothesized that human brain tissue resections can be maintained in culture with high quality and that slices are reflective of the immunoprivileged state of the healthy brain. Here, we examined both cultured and flash-frozen human brain tissue for viability, structural/functional quality and immune cell presence, and used acute rat brain slices to optimize measurements of neuronal network activity. Human brain slice cultures maintained high viability (80% or greater) for at least two weeks in culture, and the cultures showed high dendritic spine density with mature spine morphologies, as expected in the adult brain. We also optimized multi-electrode array recordings using acute rat brain slices and obtained preliminary recordings from human brain slices. Since human brain tissue donations usually come from oncology patients, we examined flash-frozen 'non-tumor' as well as a small number of 'tumor' tissues for immune cells that commonly infiltrate brain tumors and did not find large populations of these cells. Overall, our results show human brain slice cultures can be maintained at high quality for at least two weeks. Future neuroHIV studies will infect brain slices using a fluorescent HIV-1 (NL-BAL eGFP) either by directly adding virus to slice culture or by adding patient-matched, infected monocyte-derived-macrophages or T cells. Results here indicate that tissue quality is maintained throughout the entire 15-day experimental timeline and will help ensure neuroHIV studies are carried out using only high-quality brain slice cultures.
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Details
- Title
- Optimization of human organotypic brain slice cultures for use as a novel model of neuroHIV
- Creators
- James Arch Johnson
- Contributors
- Olimpia Meucci (Advisor)
- Awarding Institution
- Drexel University
- Degree Awarded
- Master of Science (M.S.)
- Publisher
- Drexel University; Philadelphia, Pennsylvania
- Number of pages
- viii, 69 pages
- Resource Type
- Thesis
- Language
- English
- Academic Unit
- Microbiology and Immunology; College of Medicine; Drexel University
- Other Identifier
- 991021819215604721