An in vitro platform for quantifying cell cycle phase lengths in primary human intestinal epithelial cells.

ABSTRACT: The intestinal epithelium dynamically controls cell cycle, yet no experimental platform exists for directly analyzing cell cycle phases in non-immortalized human intestinal epithelial cells (IECs). Here, we present two reporters and a complete platform for analyzing cell cycle phases in live primary human IECs. We interrogate the transcriptional identity of IECs grown on soft collagen, develop two fluorescent cell cycle reporter IEC lines, design and 3D print a collagen press to make chamber slides for optimal imaging while supporting primary human IEC growth, live image cell cycle dynamics, then assemble a computational pipeline building upon free-to-use programs for semi-automated analysis of cell cycle phases. The PIP-FUCCI construct allows for assigning cell cycle phase from a single i

SUBMITTER: Cotton MJ 

PROVIDER: S-EPMC11219882 | biostudies-literature | 2024 Jul

REPOSITORIES: biostudies-literature