title: A multistep procedure to prepare pre-vascularized cardiac tissue constructs using adult stem sells, dynamic cell cultures, and porous scaffolds.
abstract: The vascularization of tissue engineered products represents a key issue in regenerative medicine which needs to be addressed before the translation of these protocols to the bedside can be foreseen. Here we propose a multistep procedure to prepare pre-vascularized three-dimensional (3D) cardiac bio-substitutes using dynamic cell cultures and highly porous biocompatible gelatin scaffolds. The strategy adopted exploits the peculiar differentiation potential of two distinct subsets of adult stem cells to obtain human vascularized 3D cardiac tissues. In the first step of the procedure, human mesenchymal stem cells (hMSCs) are seeded onto gelatin scaffolds to provide interconnected vessel-like structures, while human cardiomyocyte progenitor cells (hCMPCs) are stimulated in vitro to obtain their commitment toward the cardiac phenotype. The use of a modular bioreactor allows the perfusion of the whole scaffold, providing superior performance in terms of cardiac tissue maturation and cell survival. Both the cell culture on natural-derived polymers and the continuous medium perfusion of the scaffold led to the formation of a densely packaged proto-tissue composed of vascular-like and cardiac-like cells, which might complete maturation process and interconnect with native tissue upon in vivo implantation. In conclusion, the data obtained through the approach here proposed highlight the importance to provide stem cells with complementary signals in vitro able to resemble the complexity of cardiac microenvironment.
authors.affiliation: Biomaterials Unit, International Center for Materials Nanoarchitectonics, National Institute for Materials Science Tsukuba, Japan ; International Clinical Research Center, Integrated Center of Cellular Therapy and Regenerative Medicine, St. Anne's University Hospital Brno, Czech Republic.
authors.forename: Stefania
authors.initials: S
authors.lastname: Pagliari
authors.affiliation: Interdepartmental Research Center "E. Piaggio", University of Pisa Italy ; Institute of Clinical Physiology, National Research Council (CNR) Pisa, Italy.
authors.forename: Annalisa
authors.initials: A
authors.lastname: Tirella
authors.affiliation: Interdepartmental Research Center "E. Piaggio", University of Pisa Italy ; Institute of Clinical Physiology, National Research Council (CNR) Pisa, Italy.
authors.forename: Arti
authors.initials: A
authors.lastname: Ahluwalia
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Center Leiden, Netherlands.
authors.forename: Sjoerd
authors.initials: S
authors.lastname: Duim
authors.affiliation: Department of Molecular Cell Biology, Leiden University Medical Center Leiden, Netherlands.
authors.forename: Marie-Josè
authors.initials: MJ
authors.lastname: Goumans
authors.affiliation: Biomaterials Unit, International Center for Materials Nanoarchitectonics, National Institute for Materials Science Tsukuba, Japan.
authors.forename: Takao
authors.initials: T
authors.lastname: Aoyagi
authors.affiliation: Biomaterials Unit, International Center for Materials Nanoarchitectonics, National Institute for Materials Science Tsukuba, Japan ; International Clinical Research Center, Integrated Center of Cellular Therapy and Regenerative Medicine, St. Anne's University Hospital Brno, Czech Republic.
authors.forename: Giancarlo
authors.initials: G
authors.lastname: Forte
keywords: adult stem cells; cardiac tissue engineering; dynamic culture; patient-derived stem cells; vascularized three-dimensional (3D) scaffolds