Date of Award
Bachelor of Science
The original role of astrocytes was believed to have been as a neuronal-supportive cell in the brain. It has now been discovered that they play imperative roles, from reuptake of neurotransmitters from the extracelluar space to signal propagation and developmental control by the release of factors into the extracellular space. SH-SY5Y and IMR-32 cells are common neuroblastoma cell lines which model cancerous brain cells when left undifferentiated. In recent studies tissue inhibitors of metalloproteinases (TIMPs) have been implicated in neurodegenerative diseases, but their exact role in cell death is unknown. A double-blind cell culture experiment was conducted using astrocytes from wild type and TIMP-1 knockout mice to evaluate the role of TIMP-1 in neuronal cell death. Undifferentiated and differentiated SH-SY5Y cells, as well as undifferentiated IMR-32 cells, were treated with either wild type glial cell media (WT-GCM) or TIMP-1 knockout glial cell media (KO-GCM) for 24 hours and cell viability was evaluated. Undifferentiated SH-SY5Y and IMR-32 cells exposed to WT-GCM showed a significant increase in cell death when compared to cells exposed to KO-GCM. The glial conditioned media had no effect on SH-SY5Y cells differentiated using retinoic acid. Supplementation of the KO-GCM with recombinant TIMP-1 to physiological levels had no impact on cell death in SH-SY5Y undifferentiated cells, while supplementation of WT-GCM with recombinant TIMP-1 completely blocked the cell death seen following WT-GCM treatment alone. Heating of the WT-GCM completely eliminated the increased cell death produced by the WT-GCM. These results suggest that TIMP-1 may be modulating a temperature-dependent, cell media-soluble factor that is released by astrocytes and influences cell death mechanisms of undifferentiated neuroblastoma cells.
Nicaise, Alexandra, "The Investigation of a TIMP-1-modulated Glial-Derived Factor Affecting Neuroblastoma Cell Death". Senior Theses, Trinity College, Hartford, CT 2013.
Trinity College Digital Repository, http://digitalrepository.trincoll.edu/theses/338