Investigating mitotic fidelity in rejuvenated cells

This project aims to investigate the mitotic fidelity of rejuvenated cells.

We will start by evaluating the safety of rejuvenated cells. We will expand our rejuvenation protocol to rejuvenate cells from donors of different ages and assess the rejuvenating power of our Maturation-Phase Transient Reprogramming (MPTR) method. By using old and very old donors we will be able to assess if the starting age of the donors affects the rejuvenation capacity of our method. We will subsequently use these rejuvenated cells to assess their mitotic fidelity.
We anticipate several possible outcomes from this analysis. On one hand, since we are making cells younger, it is conceivable that cells improve their mitotic segregation efficiency, implying that the age effect is predominant for mitotic fidelity. On the other hand, it is equally possible that by taking cells into pluripotency the pluripotency effect is dominant, and the rejuvenated cells will make more mistakes in mitosis. In this task we will know what biological state wins this tug-of-war – age or pluripotency – and reveal if our method of rejuvenating cells also makes them safe to use in downstream applications.
The second major aim of the project is to dissect the molecular pathways that control mitotic fidelity in rejuvenated cells. We will start by characterising key mitotic players chosen based on our own results and published studies, which are affected by either ageing or pluripotency. In parallel, we will do an unbiased screen to assess which mitotic proteins are affected by rejuvenation. Therefore, we will have the full picture of which players we can harness to make cells divide more faithfully in the future.