- Title
- An investigation of mitochondrial dynamics and networks observed within human undifferentiated and differentiated cell lines
- Creator
- Houseman, Pascalené Shannon
- ThesisAdvisor
- Prinsloo, Earl
- Subject
- Mitochondria
- Subject
- Mitochondrial pathology
- Subject
- Degeneration (Pathology)
- Subject
- Mesenchymal stem cells
- Subject
- Neural stem cells
- Subject
- Cell lines
- Subject
- Reactive oxygen species (ROS)
- Date
- 2018
- Type
- text
- Type
- Thesis
- Type
- Masters
- Type
- MSc
- Identifier
- http://hdl.handle.net/10962/60687
- Identifier
- vital:27816
- Description
- Mitochondrial dynamics refers to a series of constant division and fusion cycles that form interconnecting networks within healthy cells. Reactive oxygen species (ROS) are the byproducts of cellular redox reactions, and, when in excess, have been linked to degenerative diseases and aging. Mesenchymal stem cells (MSCs) require a niche that presents with low levels of ROS; this enables the stem cell to maintain its “sternness”, the stem cell population, as well as the ability to adhere, migrate, and proliferate. If ROS levels increase within the MSC niche, inhibition of cellular adhesion and migration occurs. In contrast, neural stem cells require a niche that presents with a high level of ROS, aiding in their proliferative, self- renewing capacities. Investigations into what constitutes a healthy mitochondrial network versus the disease state of the network are required in order to determine what promotes degeneration and aging within stem cells. It was hypothesized that increased levels of ROS would stunt the ability of MSCs to attach and migrate, and hinder their abilities of proliferation and differentiation. In contrast, neuronal differentiation would present with an increased proliferation. This led to the investigation into the effects of ROS and oxidative stress, and the resulting mitochondrial dynamics, have on undifferentiated and differentiated mesenchymal stem and SH-SY5Y cells. Upon the addition of non-lethal S3I-201 (STAT3 has been linked to a reduction in ROS) to MSCs, an increase in ROS was observed. Higher concentrations of STAT3 inhibitor resulted in a decrease in MSC attachment and proliferation. When exposed to similar conditions, the SH-SY5Y cells underwent an increased proliferation; due to multiple restrictions, they were not used any further within the study. Mitochondrial dynamics were observed using a fusion promoter (M1) and a fission inhibitor (Mdivi-1); the MSCs were dosed with varying concentrations in order to determine the effects that mitochondrial dysfunction may have on the established networks, and cell survival. The mitochondria within MSCs migrated to the extensions of the cell, and displayed an alteration in morphology, or were clustered around the nucleus and/or the lipid deposits. These high density clusters correlated with a high intensity of fluorescence using 2’,7’- dichlorofluorescein diacetate. In conclusion, varying concentrations of ROS have different effects on MSCs in terms of overall maintenance and function; mitochondrial dynamics play an important role in cell survivability and the fate of stem cell differentiation. Further investigation into the mitochondrial dynamics and networks of these cell lines and their differentiated progeny is required.
- Format
- 258 pages, pdf
- Publisher
- Rhodes University, Faculty of Science, Biotechnology Innovation Centre (RUBIC)
- Language
- English
- Rights
- Houseman, Pascalené Shannon
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