Network coupled astrocytes as a model system to study inflammatory properties
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Typ
Examensarbete för masterexamen
Master Thesis
Master Thesis
Program
Biotechnology (MPBIO), MSc
Publicerad
2015
Författare
Werner, Tony
Modellbyggare
Tidskriftstitel
ISSN
Volymtitel
Utgivare
Sammanfattning
Before antibiotics and vaccines were discovered, microbes were the primary cause of mortality rates. However, with the drastically altered life style that comes with economic development, diseases that are non-infectious, chronic and recurrent have replaced microbial diseases as the biggest challenge to human health. Cancer, cardiovascular- and neurodegenerative diseases are some examples with potentially debilitating outcomes, and the common denominator among these have been recognized to be the pathways of chronic low-grade inflammation. However, inflammation is not well understood and research must thus be conducted to better understand the condition and elucidate potential therapeutic solutions. Because of this, scientists have employed both in vivo and in vitro systems to study inflammation but because of the lack of knowledge, especially regarding neuroinflammation, in vitro assays have produced many contradictory results. A reliable in vitro system is an attractive option to in vivo systems, as it is relatively inexpensive, quick, less complex and eliminates the ethical concern for animals. The aim of this project was to develop a model cell system consisting of network coupled astrocytes and analyze changes in biomarkers when exposed to the pro-inflammatory mediators, lipopolysaccharide (LPS) and interleukin-1 beta (IL-1), for 24 hours, 3, 6 and 9 days. Analyses were done by enzyme-linked immunosorbent assay, immunofluorescence, and Western blot. The astrocyte cell cultures were investigated under different cultivation parameters, of which two different seras were tested; one of which has been shown to promote microglial proliferation. The cultures with the serum that has been shown to be beneficial to microglial proliferation had drastic effects on some of the investigated biomarkers, with a stronger induction of the toll-like receptor 4, reduced the Na+/K+-ATPase instead of an increase and an induced transcription of excitatory amino acid transporter 2. However, both seras reorganized astrocytic actin filaments similarly, caused a polymerization of globular actin, neither released the pro-inflammatory cytokines IL-1 or tumor necrosis factor alpha and in both serum groups the alteration to the biomarkers returned to control levels during longer stimulations than 24 hours. These results suggest that the astrocytes and microglia are not capable of initiating an inflammation by themselves, but microglia do appear to be at least partly responsible for observed alterations to known biomarkers. Furthermore, as previous studies have produced inflammatory reactive astrocytes by LPS or IL-1, the results imply that additional unknown factors are missing, possibly an unrecognized contaminating cell type.
Beskrivning
Ämne/nyckelord
Biologiska vetenskaper , Bioinformatik och systembiologi , Livsvetenskaper , Biological Sciences , Bioinformatics and Systems Biology , Life Science