A method for reversible permeabilization of isolated rat ventricular myocytes

Area of Science:

• Cardiology
• Cell Biology
• Biochemistry

Background:

• Isolated cardiac myocytes are crucial for studying heart function.
• Introducing molecules into cells is essential for various research applications.
• Existing methods for cardiac myocyte permeabilization can compromise cell viability.

Purpose of the Study:

• To develop and validate a method for permeabilizing and resealing rat ventricular myocytes.
• To enable the introduction of large molecules into cardiac myocytes while preserving cell viability.
• To assess the efficacy and safety of the permeabilization-resealing protocol.

Main Methods:

• Utilized Streptolysin O to create transient pores in the myocyte membrane.
• Employed fetal bovine serum to reseal the membrane pores.
• Assessed cell viability and membrane integrity using propidium iodide and fluorescein diacetate staining.
• Investigated molecular uptake using FITC-labeled dextrans and bovine serum albumin via confocal microscopy.

Main Results:

• Successfully permeabilized and resealed rat ventricular myocytes, maintaining cell viability.
• Demonstrated the successful entry and retention of large molecules (up to 148 kDa) into the cells.
• Confirmed that the protocol preserves normal myocyte morphology and electrophysiological response.
• Distinguished viable, permeabilized cells from dead and unpermeabilized cells.

Conclusions:

• The described method offers a cost-effective and simple approach for introducing large molecules into cardiac myocytes.
• This technique maintains cell viability and normal cellular function post-treatment.
• The protocol is suitable for applications requiring intracellular delivery in isolated ventricular myocytes.