title: Verification of cell viability in bioengineered tissues and organs before clinical transplantation.
abstract: The clinical outcome of transplantations of bioartificial tissues and organs depends on the presence of living cells. There are still no standard operative protocols that are simple, fast and reliable for confirming the presence of viable cells on bioartificial scaffolds prior to transplantation. By using mathematical modeling, we have developed a colorimetric-based system (colorimetric scale bar) to predict the cell viability and density for sufficient surface coverage. First, we refined a method which can provide information about cell viability and numbers in an in vitro setting: i) immunohistological staining by Phalloidin/DAPI and ii) a modified colorimetric cell viability assay. These laboratory-based methods and the developed colorimetric-based system were then validated in rat transplantation studies of unseeded and seeded tracheal grafts. This was done to provide critical information on whether the graft would be suitable for transplantation or if additional cell seeding was necessary. The potential clinical impact of the colorimetric scale bar was confirmed using patient samples. In conclusion, we have developed a robust, fast and reproducible colorimetric tool that can verify and warrant viability and integrity of an engineered tissue/organ prior to transplantation. This should facilitate a successful transplantation outcome and ensure patient safety.
authors.affiliation: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institutet, Huddinge, Stockholm, Sweden.
authors.forename: Philipp
authors.initials: P
authors.lastname: Jungebluth
authors.affiliation: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institutet, Huddinge, Stockholm, Sweden.
authors.forename: Johannes C
authors.initials: JC
authors.lastname: Haag
authors.affiliation: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institutet, Huddinge, Stockholm, Sweden.
authors.forename: Mei L
authors.initials: ML
authors.lastname: Lim
authors.affiliation: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institutet, Huddinge, Stockholm, Sweden.
authors.forename: Greg
authors.initials: G
authors.lastname: Lemon
authors.affiliation: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institutet, Huddinge, Stockholm, Sweden.
authors.forename: Sebastian
authors.initials: S
authors.lastname: Sjöqvist
authors.affiliation: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institutet, Huddinge, Stockholm, Sweden.
authors.forename: Ylva
authors.initials: Y
authors.lastname: Gustafsson
authors.affiliation: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institutet, Huddinge, Stockholm, Sweden.
authors.forename: Fatemeh
authors.initials: F
authors.lastname: Ajalloueian
authors.affiliation: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institutet, Huddinge, Stockholm, Sweden.
authors.forename: Irina
authors.initials: I
authors.lastname: Gilevich
authors.affiliation: Department of Molecular Medicine and Surgery, Division of Cardiothoracic Surgery, Karolinska Institutet, Karolinska University Hospital, Solna, Stockholm, Sweden.
authors.forename: Oscar E
authors.initials: OE
authors.lastname: Simonson
authors.affiliation: Department of Molecular Medicine and Surgery, Division of Cardiothoracic Surgery, Karolinska Institutet, Karolinska University Hospital, Solna, Stockholm, Sweden.
authors.forename: Karl H
authors.initials: KH
authors.lastname: Grinnemo
authors.affiliation: Department of Molecular Medicine and Surgery, Division of Cardiothoracic Surgery, Karolinska Institutet, Karolinska University Hospital, Solna, Stockholm, Sweden.
authors.forename: Matthias
authors.initials: M
authors.lastname: Corbascio
authors.affiliation: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institutet, Huddinge, Stockholm, Sweden.
authors.forename: Silvia
authors.initials: S
authors.lastname: Baiguera
authors.affiliation: University of Rome "Tor Vergata", Department of Industrial Engineering, Intrauniversitary Consortium for Material Science and Technology (INSTM), Research Unit "Tor Vergata", Rome, Italy.
authors.forename: Costantino
authors.initials: C
authors.lastname: Del Gaudio
authors.affiliation: Center for Bioscience, Department of Biosciences and Nutrition, Karolinska Institutet, Huddinge, Stockholm, Sweden.
authors.forename: Staffan
authors.initials: S
authors.lastname: Strömblad
authors.affiliation: Advanced Center for Translational Regenerative Medicine (ACTREM), Karolinska Institutet, Huddinge, Stockholm, Sweden. Electronic address: paolo.macchiarini@ki.se.
authors.forename: Paolo
authors.initials: P
authors.lastname: Macchiarini
mesh_terms: D000818:Animals; D021522:Bioartificial Organs; D057005:Bioengineering / Q000379:methods*; D002448:Cell Adhesion / Q000187:drug effects; D002452:Cell Count; D002470:Cell Survival / Q000187:drug effects; D005260:Female; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000166:cytology / Q000187:drug effects; D008855:Microscopy, Electron, Scanning; D011140:Polyurethanes / Q000494:pharmacology; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results; D054457:Tissue Scaffolds / Q000737:chemistry; D014132:Trachea / Q000502:physiology; D057170:Translational Research, Biomedical; D014180:Transplantation; D055815:Young Adult
chemical_list: D011140:Polyurethanes