Image-based quantification and mathematical modeling of spatial heterogeneity in ESC colonies

Area of Science:

• Cell Biology
• Systems Biology
• Developmental Biology

Background:

• Pluripotent embryonic stem cells (ESCs) can differentiate into all cell types.
• ESC pluripotency is maintained by intracellular and transcriptional factors.
• Spatial organization and cell-cell interactions in ESCs influence fate decisions, but links to transcriptional patterns are unexplored.

Purpose of the Study:

• To investigate the relationship between transcriptional patterns and spatial organization in embryonic stem cell colonies.
• To develop and apply a systems biology approach combining live-cell imaging, image analysis, and mathematical modeling.
• To quantify ESC colony morphology and spatial clustering based on gene expression levels.

Main Methods:

• Live-cell imaging and quantitative image analysis of ESC cultures.
• Development of multiscale mathematical models for ESC growth.
• Quantitative measures for morphology and spatial clustering of ESCs with varying expression levels.
• Comparison of in vitro and in silico ESC cultures.

Main Results:

• ESC colonies exhibit significant variability in morphology and spatial fluorescence patterns.
• Most ESC colonies contain a single cluster of self-renewing cells.
• These self-renewing cells are predominantly located in the interior of the colony.
• The integrated approach revealed potential transcription factor-related mechanisms driving observed patterns.

Conclusions:

• The spatial arrangement of ESCs is intrinsically linked to their transcriptional state and self-renewal capacity.
• Internal clustering of self-renewing cells is a common feature in ESC colonies.
• Systems biology approaches integrating imaging and modeling are powerful for uncovering complex cellular behaviors.