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Technology of cell sheet (CS) engineering resulted in significant advance in regenerative medicine in experimental and clinical development. Cell sheets comprise of viable cells along with extracellular matrix and can be used as a minimal construct for transplantation of cells and tissue repair. Furthermore, cells within CS can be modified by viral transduction, which is a feasible and effective way to “tune up” their therapeutic potential. Our group has extensively developed CS from adult mammalian stem cells focusing on mesenchymal stromal (MSC) and cardiac stem cells (CSC) using CS assembly without thermoresponsive dishes and viral expression of growth factors. Our findings in hindlimb ischemia model supported the concept of CS being superior to dispersed cells injection due to higher survival of cells. Furthermore, we established a method for viral expression of VEGF165 in CS and found them to exceed control CS or injected cells in angiogenesis and tissue protection. Another developed application was CS delivery of MSC or CSC to infarcted myocardium in animal models where we found them to significantly induce angiogenesis, positively influence ventricular morphology and reduce fibrosis giving them a prospect for cardiac plastics during surgical procedures. Furthermore, we addressed the “fate” of transplanted CS and found them to interact with the host undergoing vascularization and certain extent of proliferation in skeletal muscle and myocardium. Finally, we attempted to generate decellularized CS to study the role of matrix components in regenerative functions and discuss probable application of CS object for study of cellular interactions and tissue formation.