Isolation and Maintenance of Cardiac Stem Cell

One particular type of stem cell that has garnered significant attention in recent years is the cardiac stem cell. These specialized cells hold immense promise for the treatment of cardiovascular diseases, offering the potential to restore damaged heart tissue and improve overall cardiac function.

The isolation and maintenance of cardiac stem cells represent critical steps in harnessing their therapeutic potential. Creative Biolabs delves into the intricacies of the isolation and maintenance procedures for these cells, highlighting the materials required, the step-by-step procedure, and important notes to consider for successful isolation and cultivation.

Materials Required

• Cell culture media
• Fetal bovine serum (FBS)
• Antibiotics
• Trypsin-EDTA solution
• Phosphate-buffered saline (PBS)

Procedure

Animal Tissue Harvest

Obtain cardiac tissue from appropriate animal models following ethical guidelines and regulations. Ensure aseptic conditions during tissue harvest to minimize contamination. Excise the heart and place it in a sterile dish containing chilled PBS.

Tissue Dissociation

Transfer the excised heart to a petri dish and rinse it with PBS to remove excess blood. Minutely dissect the cardiac tissue into small pieces and transfer the tissue fragments to a tube containing enzyme solution. Incubate for a predetermined duration at an appropriate temperature to facilitate tissue dissociation.

Cell Extraction

Following tissue digestion, neutralize the enzyme solution using an equal volume of culture media supplemented with FBS. Pass the digested tissue suspension through a cell strainer to remove undigested tissue fragments. Centrifuge the resulting cell suspension at a low speed to separate cells from debris.

Cell Culture and Expansion

Resuspend the pelleted cells in culture media supplemented with appropriate growth factors and antibiotics. Plate the cells in cell culture flasks or dishes and place them in a controlled environment. Regularly replenish the culture media to ensure optimal nutrient supply and cell growth. Monitor cell proliferation and passage the cells at 70-80% confluency.

Notes

• Choose an animal model that closely mimics the physiological characteristics of human cardiac tissue. Consider the age and health status of the animals, as these factors can influence the quality and behavior of cardiac stem cells.
• Maintain a sterile environment throughout the procedure to prevent contamination.
• Experiment with different enzyme solutions, concentrations, and digestion times to achieve optimal tissue dissociation and maximize cell yield.
• Optimize the culture media formulation by considering the specific requirements of cardiac stem cells. Include appropriate growth factors and supplements that support stem cell maintenance and expansion.

Creative Biolabs provides an overview of the materials required, the step-by-step procedure, and important notes to consider during the isolation and cultivation of these cells. By following these guidelines and continuously refining the techniques, researchers can harness the power of cardiac stem cells to advance regenerative medicine.

For more information, please contact us.

Reference

1. Messina E, et al. Isolation and expansion of adult cardiac stem cells from human and murine heart. Circulation research, 2004, 95(9): 911-921.

For Research Use Only. Not For Clinical Use.