Evaluation of the immunomodulatory effect of phagocytosis inducers on neuronal (SH-SY5Y) cell line
Kuhlefelt, Kira (2022)
Kuhlefelt, Kira
2022
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Julkaisun pysyvä osoite on
https://urn.fi/URN:NBN:fi-fe2022052638861
https://urn.fi/URN:NBN:fi-fe2022052638861
Tiivistelmä
Ischemic stroke is a medical emergency that cuts off oxygen supply to the brain. Within minutes, brain cells start to die. Even to date, treatment options are limited, and there is a serious need for better therapeutical alternatives.
This project is a part of a larger project, where the long-term goal is to be able to alleviate and prevent brain injury after ischemic stroke, by investigating how the overexpression of genes of interest (GOIs) affects microglia and recovery after stroke. Initial results in the mouse microglia cell line BV2 have shown, that most of the GOIs induce microglial phagocytosis or high expression of pro-inflammatory mediators. The aim of my study was to determine how the expression of GOIs in neurons affect their release of inflammatory cytokines. Neuroblastoma cells, SH-SY5Y, was subcultured and transfected. The expression of proteins was determined with Western Blot, and cytokine production with enzyme-linked immunosorbent assay (ELISA). In this cell line the GOIs did not induce any cytokine production, but most of them did express the gene after transfection. On the other hand, the protein expression in Western Blot was slightly less successful. Based on this study in SH-SY5Y cells, neurons do not seem to enhance the overall inflammatory state in the brain after stroke.
This project is a part of a larger project, where the long-term goal is to be able to alleviate and prevent brain injury after ischemic stroke, by investigating how the overexpression of genes of interest (GOIs) affects microglia and recovery after stroke. Initial results in the mouse microglia cell line BV2 have shown, that most of the GOIs induce microglial phagocytosis or high expression of pro-inflammatory mediators. The aim of my study was to determine how the expression of GOIs in neurons affect their release of inflammatory cytokines. Neuroblastoma cells, SH-SY5Y, was subcultured and transfected. The expression of proteins was determined with Western Blot, and cytokine production with enzyme-linked immunosorbent assay (ELISA). In this cell line the GOIs did not induce any cytokine production, but most of them did express the gene after transfection. On the other hand, the protein expression in Western Blot was slightly less successful. Based on this study in SH-SY5Y cells, neurons do not seem to enhance the overall inflammatory state in the brain after stroke.
Kokoelmat
- 317 Farmasia [18]