Investigating the Effects of Stanniocalcin 2 in an in vitro Stroke Model
Abstract/Contents
- Abstract
- The investigation of neuroprotective agents against ischemic injury is impeded by the lack of appropriate in vitro models. An in vitro model of ischemic stroke was developed using PC12 and induced pluripotent stem cell (iPSC) derived neuronal cell cultures exposed to oxygen-glucose deprivation (OGD) followed by reoxygenation (24 hours) under atmospheric oxygen levels. Stanniocalcin 2 (STC2) is a protein expressed in the brain and has been shown to play a key role in calcium regulation. This property, along with evidence supporting STC2’s cytoprotective role in cells facing oxidative stress, has suggested it may have a neuroprotective effect during ischemic insult. Live-dead staining from initial experiments showed that iPSC-derived neuronal cells were resistant to cell death under 4, 6, and 8-hour hypoxia at 0.2 - 0.3% oxygen. However, PC12 cells plated on collagen I and collagen IV experienced significant death after OGD relative to control cells. STC2 administration at different doses (4 ng, 20 ng, and 40 ng) yielded no difference in PC12 cell viability between the OGD group and OGD groups supplemented with protein as measured by a fluorometric LDH assay. However, with an increasing dose of STC2, fewer PC12 cells died, suggesting a significant difference in cell survival may occur with the administration of a sufficient dose of STC2. Future experiments will need to assess the effects of higher concentrations of recombinant STC2 administered during OGD and reoxygenation. Stem cell resistance to ischemia-reperfusion injury, cell variability, and lifting of cells after live-dead staining may capture the lack of significant cell death seen in experiments with iPSC-derived neurons. Thus, a more robust neuron differentiation protocol will be followed in the future to produce cells with neuronal properties and less variability.
Description
| Type of resource | text |
|---|---|
| Date modified | December 5, 2022 |
| Publication date | May 5, 2022; May 2022 |
Creators/Contributors
| Author | Patel, Jainith |
|---|---|
| Thesis advisor | George, Paul |
| Thesis advisor | Dixon, Scott |
| Thesis advisor | Long, Sharon |
| Degree granting institution | Stanford University, Department of Biology |
Subjects
| Subject | Biology, Stroke, PC12 cells, iPSC-derived neurons, Oxygen and glucose deprivation |
|---|---|
| Genre | Text |
| Genre | Thesis |
Bibliographic information
Access conditions
- License
- This work is licensed under a Creative Commons Attribution Non Commercial 4.0 International license (CC BY-NC).
Preferred citation
- Preferred citation
- Patel, J. and George, P. (2023). Investigating the Effects of Stanniocalcin 2 in an in vitro Stroke Model. Stanford Digital Repository. Available at https://purl.stanford.edu/td400md3746
Collection
Undergraduate Theses, Department of Biology, 2021-2022
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- Contact
- jjpatel@stanford.edu
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