Segmentation and tracking of stem cells in time lapse microscopy to quantify dynamic behavioral changes during...

Area of Science:

• Biotechnology
• Cell Biology
• Regenerative Medicine

Background:

• Dynamic behavior of stem cells during in vitro culture is diverse, with limited focus on cell aggregation.
• Cell aggregation, particularly in forming 3D spheroids, has been linked to enhanced proliferation and maintained stemness marker gene expression in adult human mesenchymal stem cells.

Purpose of the Study:

• To investigate the dynamic behavioral changes of stem cells triggered by 3D spheroid formation.
• To develop and validate a systematic image analysis method for quantifying stem cell behavior and stemness during in vitro culture.

Main Methods:

• A hybrid-thresholding technique for efficient segmentation of cell clusters.
• A cell tracking method based on pair-matching with topological constraints.
• Derivation of morphological indices to track 3D spheroid formation and measurement of cell motility indices for comparison between single cells and spheroids.

Main Results:

• The developed image processing techniques achieved over 90% correspondence with manual tracking.
• Analysis revealed a significant increase in cell motility associated with 3D spheroid formation.
• The findings are consistent with previous studies using gene expression approaches.

Conclusions:

• A systematic image analysis platform was proposed and validated for quantifying dynamic stem cell behavior during in vitro culture.
• The study demonstrates that 3D spheroid formation significantly influences stem cell motility.
• The developed methods can aid in evaluating stemness during cell culture through dynamic behavior analysis.