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Question: You hypothesise that EV cargo from aged cells can play a role in changing the epigenetic landscape of stem cells. Outline an experimental design that would enable you to explore the influence of EVs on your chosen epigenetic mark.
1. Isolate and culture primary mouse embryonic stem cells (mESCs). 2. Collect and purify EVs from aged cells. 3. Treat mESCs with EVs from aged cells and control cells with EVs from young cells. 4. Measure global epigenetic changes in both sets of cells. 5. Analyze the data to determine differences in the global epigenetic landscape and isolate specific epigenetic marks. 6. Analyze the data to determine if there is an influence of EVs from aged cells on epigenetic marks. 7. If an influence is observed, identify the molecular pathways by which EVs from aged cells are exerting their effects. 8. Validate the effects of EVs from aged cells on epigenetic marks in vivo by generating animal models and investigating the effects of treatment with EVs from aged cells compared to control animals not treated with EVs from aged cells.
Jan. 16, 2023, 4:35 a.m.
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