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Shriners Hospital for Children

8 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

A Contusion Model of Severe Spinal Cord Injury in Rats

Vibhor Krishna¹, Hampton Andrews¹, Xing Jin², Jin Yu¹, Abhay Varma¹, Xuejun Wen³, Mark Kindy¹

¹Department of Neuroscience, Division of Neurosurgery, Medical University of South Carolina, ²Bioengineering, Clemson University, ³Clemson-MUSC Bioengineering Joint Program

A contusion model of severe spinal cord injury is described. Detailed pre-operative, operative and post-operative steps are described to obtain a consistent model.

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).

JoVE Core

Elevated Plus Maze Test Combined with Video Tracking Software to Investigate the Anxiolytic Effect of Exogenous Ketogenic Supplements

Csilla Ari¹, Dominic P. D’Agostino^2,3, David M. Diamond^1,2, Mark Kindy^4,5,6, Collin Park¹, Zsolt Kovács⁷

¹Department of Psychology, Hyperbaric Neuroscience Research Laboratory, University of South Florida, ²Department of Molecular Pharmacology and Physiology, Metabolic Medicine Research Laboratory, Morsani College of Medicine, University of South Florida, ³Institute for Human and Machine Cognition, ⁴Department of Pharmaceutical Sciences, College of Pharmacy, University of South Florida, ⁵James A. Haley VA Medical Center, ⁶Shriners Hospital for Children, ⁷Savaria Department of Biology, ELTE Eötvös Loránd University

Here, we present a protocol to investigate changes in the anxiety level of rodent animal models. The elevated plus maze (EPM) test, used together with a video tracking software, provides a reliable method to document the effect of various potential anxiolytic treatments in preclinical laboratory scenarios.

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.