Shriners Children's Boston

9 ARTICLES PUBLISHED IN JoVE

Bioengineering

Decellularization and Recellularization of Whole Livers

Basak E. Uygun¹, Gavrielle Price¹, Nima Saeidi¹, Maria-Louisa Izamis¹, Tim Berendsen¹, Martin Yarmush¹, Korkut Uygun¹

¹Center for Engineering in Medicine, Massachusetts General Hospital, Harvard Medical School, Shriners Hospitals for Children

Perfusion decellularization is a novel technique to produce whole liver scaffolds that retains the organ's extracellular matrix composition and microarchitecture. Herein, the method of preparing whole organ scaffolds using perfusion decellularization and subsequent repopulation with hepatocytes is described. Functional and transplantable liver grafts can be generated using this technique.

Medicine

Ex Situ Normothermic Machine Perfusion of Donor Livers

Negin Karimian^1,2, Alix P.M. Matton^1,2, Andrie C. Westerkamp^1,2, Laura C. Burlage^1,2, Sanna op den Dries^1,2, Henri G.D. Leuvenink², Ton Lisman^1,2, Korkut Uygun³, James F. Markmann⁴, Robert J. Porte^1,2

¹Section of Hepato-Pancreato-Biliary Surgery and Liver Transplantation, University of Groningen, University Medical Center Groningen, ²Surgical Research Laboratory, Department of Surgery, University of Groningen, University Medical Center Groningen, ³Center of Engineering in Medicine/Surgical Services, Massachusetts General Hospital, Harvard Medical School, and Shriners Burns Hospital, ⁴Division of Transplantation, Department of Surgery, Massachusetts General Hospital, Harvard Medical School

Here we present a protocol describing oxygenated ex situ machine perfusion of donor liver grafts. This article contains a step by step protocol to procure and prepare the liver graft for machine perfusion, prepare the perfusion fluid, prime the perfusion machine and perform oxygenated normothermic machine perfusion of the liver graft.

Medicine

Functional Human Liver Preservation and Recovery by Means of Subnormothermic Machine Perfusion

Bote G. Bruinsma^*1, James H. Avruch^*2, Pepijn D. Weeder¹, Gautham V. Sridharan¹, Basak E. Uygun¹, Negin G. Karimian¹, Robert J. Porte³, James F. Markmann², Heidi Yeh², Korkut Uygun¹

¹Center for Engineering in Medicine, Dept. of Surgery, Massachusetts General Hospital, Harvard Medical School, ²Transplant Center, Dept. of Surgery, Massachusetts General Hospital, Harvard Medical School, ³Section of Hepatobiliary Surgery and Liver Transplantation, Department of Surgery, University Medical Center Groningen, University of Groningen

We describe a method of ex vivo machine perfusion of human liver grafts at subnormothermic temperature (21 °C).

Bioengineering

Bulk Droplet Vitrification for Primary Hepatocyte Preservation

Reinier J. de Vries^1,2,3, Peony D. Banik^1,2, Sonal Nagpal^1,2, Lindong Weng^1,2, Sinan Ozer^1,2, Thomas M. van Gulik³, Mehmet Toner^1,2, Shannon N. Tessier^1,2, Korkut Uygun^1,2

¹Center for Engineering in Medicine, Harvard Medical School, Massachusetts General Hospital, ²Shriners Hospitals for Children, Boston, ³Department of Surgery, University of Amsterdam

This manuscript describes an ice-free cryopreservation method for large quantities of rat hepatocytes whereby primary cells are pre-incubated with cryoprotective agents at a low concentration and vitrified in large droplets.

Immunology and Infection

Partial Heterotopic Hindlimb Transplantation Model in Rats

Marion Goutard^1,2,3, Mark A. Randolph^1,2,3, Corentin B. Taveau^1,2,3,4, Elise Lupon^1,2,3, Laurent Lantieri⁴, Korkut Uygun^1,2,3, Curtis L. Cetrulo Jr.^*1,2,3, Alexandre G. Lellouch^*1,2,3,4

¹Division of Plastic and Reconstructive Surgery, Massachusetts General Hospital, Harvard Medical School, ²Vascularized Composite Allotransplantation Laboratory, Center for Transplantation Sciences, Massachusetts General Hospital, Harvard Medical School, ³Shriners Hospital for Children, ⁴Service de Chirurgie Plastique, Hôpital Européen Georges Pompidou, Assistance Publique-Hôpitaux de Paris (APHP), Université de Paris

This paper presents a partial heterotopic osteomyocutaneous flap transplantation protocol in rats and its potential outcomes in the mid-term follow-up.

Bioengineering

A Reliable Porcine Fascio-Cutaneous Flap Model for Vascularized Composite Allografts Bioengineering Studies

Victor Pozzo^1,2,4, Golda Romano^1,2,4, Marion Goutard^1,2,4, Elise Lupon^1,2,4, Pierre Tawa^1,2,4, Aylin Acun^3,4,5, Alec R. Andrews², Corentin B. Taveau^1,2,4,6, Basak E. Uygun^1,2,3,4, Mark A. Randolph^1,2,4, Curtis L. Cetrulo^1,2,4, Alexandre G. Lellouch^1,2,4,6

¹Division of Plastic and Reconstructive Surgery, Massachusetts General Hospital, Harvard Medical School, ²Vascularized Composite Allotransplantation Laboratory, Center for Transplantation Sciences, Massachusetts General Hospital, Harvard Medical School, ³Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, ⁴Shriners Hospital for Children, ⁵Department of Biomedical Engineering, Widener University, ⁶Service de Chirurgie Plastique, Hôpital Européen Georges Pompidou, Assistance Publique-Hôpitaux de Paris (APHP), Université Paris Descartes

The present protocol describes the porcine fascio-cutaneous flap model and its potential use in vascularized composite tissue research.

Immunology and Infection

The Superficial Inferior Epigastric Artery Axial Flap to Study Ischemic Preconditioning Effects in a Rat Model

Yanis Berkane^*1,2,3,5, Austin Alana Shamlou^*1,2,4,5, Jose Reyes^1,2, Hyshem H. Lancia^1,2,5, Irina Filz von Reiterdank^1,2,5,6,7, Nicolas Bertheuil³, Basak E. Uygun^2,5,6, Korkut Uygun^1,2,5,6, William G. Austen Jr.^2,4, Curtis L. Cetrulo Jr.^1,2,4,5, Mark A. Randolph^1,2,4,5, Alexandre G. Lellouch^1,2,5,6,8

¹Vascularized Composite Allotransplantation Laboratory, Massachusetts General Hospital, ²Harvard Medical School, ³Department of Plastic, Reconstructive and Aesthetic Surgery, Rennes University Hospital Center (CHU de Rennes), University of Rennes 1, ⁴Plastic Surgery Research Laboratory, Massachusetts General Hospital, ⁵Shriners Children's Boston, ⁶Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, ⁷Department of Plastic Surgery and Hand Surgery, University Medical Center Utrecht, ⁸Department of Plastic, Reconstructive, and Aesthetic Surgery, Groupe Almaviva Santé, Clinique de l'Alma

This protocol describes harvesting, suturing, and monitoring fasciocutaneous flaps in rats that allow for good visualization and manipulation of blood flow through the superficial inferior epigastric vessels by means of clamping and ligating the femoral vessels. This is critical for studies involving ischemic preconditioning.

Bioengineering

Modified Langendorff Perfusion for Extended Perfusion Times of Rodent Cardiac Grafts

Casie A. Pendexter^1,2, Maya Bolger-Chen^1,2, Manuela Lopera Higuita^1,2, Stephanie E. J. Cronin^1,2, S. Alireza Rabi³, Asishana A. Osho³, Shannon N. Tessier^1,2

¹Center for Engineering in Medicine and Surgery, Harvard Medical School and Massachusetts General Hospital, ²Shriners Children's Boston, ³Division of Cardiac Surgery, Corrigan Minehan Heart Center, Massachusetts General Hospital

This article demonstrates the feasibility of achieving longer perfusion times (4 h) of murine cardiac grafts without function loss by employing lower (30-35 mmHg) than physiological (60-80 mmHg) perfusion pressures during Langendorff.

Medicine

Establishing a Swine Model to Study Uterus Dynamic Preservation and Transplantation

Lucile Cabanel^1,2,3, Haïzam Oubari^1,2,3, Ludivine Dion⁴, Vincent Lavoué⁴, Mark A. Randolph^2,3, Curtis L. Cetrulo^2,3,5, Korkut Uygun^1,2, Alexandre G. Lellouch^1,2,3,5, Yanis Berkane^1,2,6,7, Basak E. Uygun^1,2

¹Center for Engineering in Medicine and Surgery, Massachusetts General Hospital, Harvard Medical School, ²Shriners Children's Boston, ³Plastic Surgery Research Laboratory, Department of plastic and reconstructive surgery, Massachusetts General Hospital, Harvard Medical School, ⁴Department of Gynecology and Obstetrics, CHU de Rennes, University of Rennes, ⁵Division of Plastic Surgery Research, Vascularized Composite Allotransplantation Program Development, Cedars-Sinai Hospital, ⁶Department of Plastic, Reconstructive and Aesthetic Surgery, CHU de Rennes, University of Rennes, ⁷SITI Laboratory, UMR1236 INSERM, Établissement Français du Sang, University of Rennes

A detailed and reproducible swine uterus model is described, from surgical procurement to the initiation of machine perfusion, allowing for the study of uterus preservation in transplantation.