Time-lapse imaging of in vitro myogenesis using atomic force microscopy.

Area of Science:

• Cell Biology
• Biophysics
• Regenerative Medicine

Background:

• Myoblast therapy requires successful skeletal muscle stem cell integration.
• Myogenesis, the process of muscle formation, is crucial for therapeutic applications.
• Understanding cell division and fusion mechanisms is fundamental.

Purpose of the Study:

• To investigate membrane and actin cytoskeleton rearrangements during C2C12 myoblast fusion.
• To compare membrane dynamics during cell fusion versus cell division (cytokinesis).

Main Methods:

• Atomic Force Microscopy (AFM) for live cell imaging of membrane structure.
• Confocal Laser Scanning Microscopy (CLSM) for actin cytoskeleton visualization.

Main Results:

• AFM revealed membrane tubules forming rapidly upon cell-cell contact during myoblast fusion.
• These tubules increase in size as fusion progresses.
• CLSM showed actin filaments nucleating at contact points, forming the scaffold for these tubules, which are absent in cytokinesis.

Conclusions:

• The study elucidates the role of actin-based membrane tubules in myoblast fusion.
• Findings enhance understanding of the distinct membrane dynamics in cell division and fusion.
• This knowledge is vital for advancing myoblast-based therapies.