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Exosomes are small extracellular vesicles that have emerged as an important tool for intercellular communication. In the central nervous system, exosomes can mediate glia and neuronal communication. Once released from the donor cell, exosomes can act as discrete vesicles and travel to distant and proximal recipient cells to alter cellular function. Microglia cells secrete exosomes due to stress stimuli of alcohol abuse. The goal of this study was to investigate the effects of alcohol exposure on the biogenesis and composition of exosomes derived from microglia cell line BV-2. The BV-2 cells were cultured in exosome-free media and were either mock treated (control) or treated with 50 mM or 100 mM of alcohol for 48 and 72 h. Our results demonstrated that alcohol significantly impacted BV-2 cell morphology, viability, and protein content. Most importantly, our studies revealed that exosome biogenesis and composition was affected by alcohol treatment.
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Evaluating PC12 Cell Differentiation Following Exosome Treatments
Exosomes are extracellular vesicles ranging in size from 40 to 100nm that are used for extracellular communication to control the cell niche environment with regards to differentiation. Initial studies in our lab have shown that exosomes isolated from differentiating neurites can cause differentiation in cells absent typical neuronal hormones. However, these studies only used exosomes isolated after complete differentiation. To further clarify the role of exosomes in cell differentiation, we isolated exosomes from NGF-treated PC12 cells after two, four and six days. The exosomes were taken at the specific times due to a slight majority of cells showing differentiation at day three, with an overwhelming majority showing differentiation at day six. Day two, four and six were thus good markers in the development of cell differentiation. Each set of exosomes was added to a different well of untreated cells, which were then observed for differentiation daily over a six-day period. The results showed a considerably larger amount of percent differentiation in cells treated with the day six exosomes compared to cells treated by the day two and day four exosomes. The change in number of differentiated cells in different exosome treatments indicate that exosomal contents change over time. In an effort to make sure that no NGF contamination occurred in our treatments, an NGF-ELISA was completed testing all the treatment exosomes with results showing no NGF. Additionally, exosomes were isolated from NGF spiked media to show that no NGF was carried over in our exosome isolation process. Both tests further evidence that our exosomes did not contain NGF. Our overall results indicate that the isolated exosomes changed contents over time and that their contents caused greater percent differentiation over time. Funding provided by the Cell Biology Education Consortium (CBEC) and through the AR-EPSCoR Center for Advanced Surface Engineering.
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- Award ID(s):
- 1827066
- PAR ID:
- 10175903
- Date Published:
- Journal Name:
- Proceedings of The National Conference On Undergraduate Research (NCUR) 2019 Kennesaw State University Kennesaw, Georgia April 11-13, 2019
- Format(s):
- Medium: X
- Sponsoring Org:
- National Science Foundation
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