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Shriners Children's Boston

8 ARTICLES PUBLISHED IN JoVE

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Bioengineering

Decellularization and Recellularization of Whole Livers
Basak E. Uygun 1, Gavrielle Price 1, Nima Saeidi 1, Maria-Louisa Izamis 1, Tim Berendsen 1, Martin Yarmush 1, Korkut Uygun 1
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.

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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. Leuvenink 2, Ton Lisman 1,2, Korkut Uygun 3, James F. Markmann 4, 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.

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Medicine

Functional Human Liver Preservation and Recovery by Means of Subnormothermic Machine Perfusion
Bote G. Bruinsma *1, James H. Avruch *2, Pepijn D. Weeder 1, Gautham V. Sridharan 1, Basak E. Uygun 1, Negin G. Karimian 1, Robert J. Porte 3, James F. Markmann 2, Heidi Yeh 2, Korkut Uygun 1
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).

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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 Gulik 3, 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.

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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 Lantieri 4, 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.

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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. Andrews 2, 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.

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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 Bertheuil 3, 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.

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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 Rabi 3, Asishana A. Osho 3, 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.

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