Encapsulation of Pediatric Cardiac-Derived C-Kit+ Cells in Cardiac Extracellular Matrix Hydrogel for Echocardiography-Directed Intramyocardial Injection in Rodents.

Pediatric cardiac-derived c-kit+ cell therapies represent an innovative approach for cardiac tissue repair that have demonstrated promising improvements in recent studies and offer multiple benefits, such as easy isolation and autologous transplant. However, concerns about failure of engraftment and transient paracrine effects have thus far limited their use. To overcome these issues, an appropriate cell delivery vehicle such as a cardiac extracellular matrix (cECM) hydrogel can be utilized. This naturally derived biomaterial can support embedded cells, allowing for local diffusion of paracrine factors, and provide a healthy microenvironment for optimal cellular function. This protocol focuses on combining cardiac-derived c-kit+ cells and a cECM hydrogel to prepare a minimally invasive, dual therapeutic for in vivo delivery. We also outline a detailed method for ultrasound-guided intramyocardial injection of cell-laden hydrogels in a rodent model. Additional steps for labeling cells with a fluorescent dye for in vivo cell tracking are provided.