Original Publication Date
The Biophysical Journal
DOI of Original Publication
Date of Submission
Somatic cell reprogramming to pluripotency requires an immediate increase in cell proliferation and reduction in cell size. It is unknown whether proliferation and biomass controls are similarly coordinated with early events during the differentiation of pluripotent stem cells (PSCs). This impasse exists because PSCs grow in tight clusters or colonies, precluding most quantifying approaches. Here, we investigate live cell interferometry as an approach to quantify the biomass and growth of HSF1 human PSC colonies before and during retinoic acid-induced differentiation. We also provide an approach for measuring the rate and coordination of intracolony mass redistribution in HSF1 clusters using live cell interferometry images. We show that HSF1 cells grow at a consistent, exponential rate regardless of colony size and display coordinated intracolony movement that ceases with the onset of differentiation. By contrast, growth and proliferation rates show a decrease of only ∼15% decrease during early differentiation despite global changes in gene expression and previously reported changes in energy metabolism. Overall, these results suggest that cell biomass and proliferation are regulated independent of pluripotency during early differentiation, which is distinct from what occurs with successful reprogramming.
From The Biophysical Journal, Zangle, T.A., Chun, J., Zhang, J., et al., Quantification of Biomass and Cell Motion in Human Pluripotent Stem Cell Colonies, Vol. 105, Page 593, Copyright © 2013 Biophysical Society. Published by Elsevier Inc. Reprinted with permission.
Is Part Of
VCU Physics Publications
Analysis section and eight figures.
mmc2.mp4 (2524 kB)
Colony Motion and LCI Time-Lapse Measurements of Mass Distributions: One Untreated (Control) Colony
mmc3.mp4 (2284 kB)
Colony Motion and LCI Time-Lapse Measurements of Mass Distributions: One Untreated (Control) Colony for 45 Min.
mmc4.mp4 (3922 kB)
Colony Motion and LCI Time-Lapse Measurements of Mass Distributions: Colony from Day 4 of RA Treatment over a period of 80 Min.
mmc5.mp4 (1727 kB)
Colony Motion and LCI Time-Lapse Measurements of Mass Distributions: Colony from Day 6 of RA Treatment over a Period of 50 Mins.