or less. My Hypothesis is that EV cargo from aged cells can play a role in changing the epigenetic landscape of stem cells, and that this can be tracked and quantified. I predict that there will be

. Hypothesis: EVs released from aged cells can alter the epigenetic landscape of stem cells, resulting in changes to the genomic imprinting. Rationale: Genomic imprinting is an epigenetic phenomenon where certain genes are expressed differently depending on which parent they are inherited from. It has been suggested that there is a link between aging and epigenetic modifications, which may affect the expression of imprinted genes. The release of EVs from aged cells has the potential to influence the epigenetic landscape of stem cells, and consequently the expression of imprinted genes. Therefore, it is proposed that EVs can have a role in altering the epigenetic landscape of stem cells, resulting in changes to genomic imprinting. Methodology: To test this hypothesis, a cell culture system will be used to compare the effects of EVs released from aged cells and EVs released from young cells. This will involve generating a population of aged cells, which will be derived from a parental cell line with an established genomic imprinting pattern. These aged cells will then be incubated in culture for a period of time and their EVs will be isolated and collected. A population of young cells will also be generated from the same parental cell line, and the EVs from these young cells will also be isolated and collected. The EVs from both the aged and young cells will then be added to a population of stem cells, and the effects on the expression of the imprinted genes will be compared. The expression of the imprinted genes will be assessed using quantitative PCR (qPCR). Quantification: The qPCR will be used to quantify the expression of the imprinted genes in order to determine if there is a difference in the expression of these genes in the presence of EVs from aged and young cells. The expression of the imprinted genes will be assessed in both the treated and untreated stem cells. Interpretation: The results of the qPCR will be used to interpret the effects of the EVs on the expression of the imprinted genes. The results will be compared between the cells treated with EVs from aged and young cells to determine if there is a difference in the expression of the imprinted genes. If there is a difference in the expression of the imprinted genes, then it can be concluded that EVs can have a role in altering the epigenetic landscape of stem cells, resulting in changes to the genomic imprinting. Conclusion: This experimental design will allow for