Weaning and Adaptation of SH-SY5Y Cells to Low Glucose Media for Manganese Exposure Studies

Hannah Smerker

Abstract


SH-SY5Y cells are a dopaminergic cell line commonly used as a model for neurodegenerative diseases. Parkinson’s disease is characterized by a loss of dopamine signaling as a result of dopaminergic cell death within the substantia nigra. Manganism is an idiopathic Parkinsonism-like disease caused by an accumulation of manganese in cells of the basal ganglia and presents symptoms similar to Parkinson’s. Extracellular dopamine levels are decreased in SH-SY5Y cells exposed to manganese. Several potential mechanisms may explain the effects of manganese on dopamine levels, including changes in dopamine metabolism. Growth of SH-SY5Y cells in high glucose Dulbecco’s Modified Eagles Medium (DMEM) prevented us from discerning the levels of other dopamine metabolites using high performance liquid chromatography (HPLC) with electrochemical detection. The objective of this experiment was to determine if SH-SY5Y cells could be weaned off of DMEM high glucose media and adapted to DMEM low glucose or EMEM/Ham’s F12 media, without altering cell viability. A sequential method was developed to adapt cells. The number of days between passages was recorded, and prior to each cell passage, a trypan blue exclusion assay was conducted to measure cell viability. SH-SY5Y cells were successfully adapted to EMEM/Ham’s F12 media, whereas cell proliferation was hindered when grown in DMEM low glucose media. Cells grown in DMEM low-glucose media appeared to lose their morphology and detach from the growth surface. SH-SY5Y cells grown in EMEM/Ham’s F12 media appeared larger in size compared to cells grown in DMEM high-glucose media. Ongoing studies are quantitating differences in cell size and examining the HPLC profile for EMEM/Ham’s F12 media. If dopamine metabolite levels can be discerned, EMEM/Ham’s F12 media may allow further analysis of dopamine metabolism in SH-SY5Y cells following manganese exposure for use as a model of neurodegenerative diseases.


Keywords


SH-SY5Y; dopaminergic; glucose; dopamine; cell viability; cell culture

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