A New Microengineered Platform for 4D Tracking of Single Cells in a Stem-Cell-Based In Vitro Morphogenesis Model

Area of Science:

• Developmental Biology
• Stem Cell Biology
• Bioengineering

Background:

• Stem cell-based in vitro models are crucial for understanding embryonic patterning.
• Traditional cell culture methods offer limited control and imaging capabilities for studying morphogenesis.
• Advanced microengineered tools can enhance control, culture duration, and data acquisition for biological models.

Purpose of the Study:

• To describe a new microengineered platform for culturing, manipulating, and analyzing in vitro morphogenesis.
• To develop a pipeline for quantifying 3D cell movement and migration within multicellular clusters using 4D imaging.
• To investigate the mechanisms of morphogenesis in stem cell aggregates.

Main Methods:

• Development of thin polycarbonate film-based microdevices for cell culture.
• Implementation of a 4D image acquisition system for tracking cell movement.
• Utilizing open-source software for quantitative analysis of cell migration in 3D.
• Culturing P19C5 mouse stem cell line and mouse embryonic stem cells (mESCs).

Main Results:

• The platform successfully supports the study of in vitro morphogenesis in non-adherent stem cell cultures.
• Observed symmetry breaking and axial elongation events in stem cell aggregates mimic early embryonic development.
• Localized cell proliferation and coordinated cell migration were identified as key drivers of P19C5 aggregate elongation.
• Stem cell aggregate polarization and elongation were found to be dependent on directed cell migration.

Conclusions:

• The developed microengineered platform provides enhanced control and imaging for studying stem cell morphogenesis.
• The findings elucidate the roles of cell proliferation and migration in embryonic patterning.
• This research offers new insights into the fundamental mechanisms governing early embryonic development using in vitro models.