Approaches to Determine Nuclear Shape in Cells During Migration Through Collagen Matrices

Area of Science:

• Cell Biology
• Biophysics
• Biomaterials

Background:

• Fibrillar collagen is a key component of the extracellular matrix (ECM) in interstitial tissues.
• Interstitial cell migration, crucial for physiological and pathological processes like cancer metastasis, often involves navigating confined spaces within the ECM.
• Nuclear deformation during cell migration can affect nuclear integrity, making nuclear shape analysis valuable for understanding biological processes.

Purpose of the Study:

• To present a protocol for an in vitro cell-collagen model.
• To detail the use of confocal microscopy for visualizing cell nuclei during migration.
• To introduce computational tools for quantifying nuclear shape dynamics.

Main Methods:

• Development of an in vitro cell-collagen model.
• Confocal microscopy for static and dynamic visualization of labeled cell nuclei.
• Two custom scripts for semi-automated quantification of nuclear shape descriptors (e.g., aspect ratio, circularity, nuclear irregularity index) and their temporal changes.

Main Results:

• Successful generation and visualization of migratory cells within a 3D collagen matrix.
• Semi-automated quantification of static and dynamic nuclear shape parameters.
• Demonstration of the method's utility by analyzing nuclear shape changes in cells migrating through collagen with and without a collagen degradation inhibitor.

Conclusions:

• The presented pipeline provides a robust method for analyzing nuclear shape changes during cell migration in a collagenous environment.
• This approach offers valuable insights into cell mechanics and nuclear integrity during migration.
• The methodology is adaptable to various cell migration assays, including 3D microfluidics and in vivo studies.