Cumulative labelling with thymidine analogues when the steady-state assumption is violated

TL;DR

This paper uses computational modeling to show that cumulative labelling with thymidine analogues overestimates cell cycle parameters when the steady-state assumption is not met, highlighting the importance of model-based interpretation.

Contribution

It demonstrates how violating the steady-state assumption affects the accuracy of cumulative labelling estimates and provides insights into experimental discrepancies.

Findings

Cumulative labelling overestimates growth fraction in non-steady populations

Model results explain discrepancies in oligodendrocyte precursor cell measurements

Highlights the importance of computational modeling in experimental design

Abstract

We present modelling results that examine the consequences of implementing cumulative labelling with thymidine analogues, to estimate the cell cycle time and growth fraction of dividing cells, when the steady-state assumption is violated. We fix the value of the cell cycle time a priori and examine whether cumulative labelling can reproduce this value. We find that the cumulative labelling technique systematically overestimates the growth fraction and cell cycle time in non-steady cell populations. Our results suggest an explanation for discrepancies in experimental measurements of oligodendrocyte precursor cell properties using cumulative labelling. These results also emphasise the utility of using computational models to determine what violating the assumptions of experimental techniques would look like in the laboratory before experiments are undertaken.