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Children’s Hospital Boston

10 ARTICLES PUBLISHED IN JoVE

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Biology

Transplantation of Whole Kidney Marrow in Adult Zebrafish
Jocelyn LeBlanc 1, Teresa Venezia Bowman 1, Leonard Zon 1
1Children's Hospital, Harvard Stem Cell Institute, Harvard Medical School

In this article, we demonstrate a method to perform HCT in adult zebrafish.

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Biology

Microinjection of Zebrafish Embryos to Analyze Gene Function
Jonathan N. Rosen 1,2, Michael F. Sweeney 1, John D. Mably 1,2
1Department of Genetics, Harvard Medical School, 2Department of Cardiology, Children’s Hospital Boston

This video shows how morpholino or mRNA can be injected into zebrafish embryos at the one-cell stage to decrease or increase the level of specific gene products during subsequent development.

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Biology

Fabrication of Myogenic Engineered Tissue Constructs
Christina A. Pacak 1,2, Douglas B. Cowan 1,2
1Department of Anesthesiology, Children's Hospital Boston and Harvard Medical School, 2Perioperative and Pain Medicine, Children's Hospital Boston and Harvard Medical School

Here, we demonstrate fabrication of collagen-based, tissue constructs containing skeletal myoblasts. These 3-D engineered constructs may be used to replace or repair tissues in vivo. For our purposes, we have designed these as an atrioventricular electrical conduit for the repair of complete heart block[1].

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Biology

Optical Mapping of Langendorff-perfused Rat Hearts
Bjoern Sill 1, Peter E. Hammer 1,2, Douglas B. Cowan 1
1Department of Anesthesiology, Perioperative and Pain Medicine, Children's Hospital Boston and Harvard Medical School, 2Departments of Cardiac Surgery, Children's Hospital Boston and Harvard Medical School

This article describes a high temporal and spatial resolution technique to optically image action potential movement on the surface of Langendorff-perfused rat hearts using a potentiometric dye (di-8-ANEPPS).

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Biology

Implantation of Engineered Tissue in the Rat Heart
Bjoern Sill 1, Ivan V. Alpatov 2, Christina A. Pacak 2, Douglas B. Cowan 2
1Department of Anesthesiology, Perioperative and Pain Medicine, Children's Hospital Boston and Harvard Medical School, 2Department of Anesthesiology, Perioperative and Pain Medicine, Children’s Hospital Boston

Here, we describe a cardiac surgical procedure to implant engineered tissue in the atrioventricular (AV)-groove of an adult Lewis rat.

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Biology

Making MR Imaging Child's Play - Pediatric Neuroimaging Protocol, Guidelines and Procedure
Nora M. Raschle 1,2, Michelle Lee 1, Roman Buechler 1, Joanna A. Christodoulou 3, Maria Chang 1, Monica Vakil 1, Patrice L. Stering 1, Nadine Gaab 1,3,4
1Department of Developmental Medicine, Children’s Hospital Boston, 2Department of Neuropsychology, University of Zurich, 3Graduate School of Education, Harvard, 4Harvard Medical School

Despite an increase in the use of structural and functional magnetic resonance imaging (fMRI) in humans, the study of young pediatric populations remains a challenge. We present a hands-on, step-by-step video protocol including guidelines for clinicians and researchers intending to perform (f)MRI in young children.

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Biology

Retro-orbital Injection in Adult Zebrafish
Emily K. Pugach 1,2, Pulin Li 1,2, Richard White 1,2,3, Leonard Zon 1,2
1Department of Hematology and Oncology, Children’s Hospital Boston, 2Harvard Stem Cell Institute, Harvard Medical School, 3Department of Medical Oncology, Dana Farber Cancer Institute

Here we show how to do retro-orbital injection in adult zebrafish.

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Immunology and Infection

Induction of Alloantigen-specific Anergy in Human Peripheral Blood Mononuclear Cells by Alloantigen Stimulation with Co-stimulatory Signal Blockade
Jeff K. Davies 1,2, Christine M. Barbon 1, Annie R. Voskertchian 1, Lee M. Nadler 1,2, Eva C. Guinan 3,4
1Medical Oncology, Dana Farber Cancer Institute, 2Department of Medicine, Brigham and Womens Hospital, 3Pediatric Oncology, Dana Farber Cancer Institute, 4Division of Hematology/Oncology, Children’s Hospital Boston

This paper describes a simple technique to induce alloantigen-specific anergy in human peripheral blood mononuclear cells. The technique can be applied clinically to generate non-alloreactive donor cells. Infusion of these cells could improve immune reconstitution and reduce toxicity after allogeneic hematopoietic stem cell transplantation.

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Bioengineering

An Improved Method for the Preparation of Type I Collagen From Skin
Christina A. Pacak 1,2, Allison A. MacKay 1, Douglas B. Cowan 1,2
1Department of Anesthesiology, Perioperative and Pain Medicine, Boston Children's Hospital, 2Department of Anesthesia, Harvard Medical School

Traditional procedures for the isolation of soluble type 1 collagen (COL1) require about 10 days from start to finish because of lengthy buffer incubations and laborious resuspensions of fibrils. Here, we describe a means to purify COL1 from small dermal biopsies in less than 3 hr.

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Biology

Rapid Isolation And Purification Of Mitochondria For Transplantation By Tissue Dissociation And Differential Filtration
Janine M. Preble 1, Christina A. Pacak 2, Hiroshi Kondo 1, Allison A. MacKay 2, Douglas B. Cowan 2, James D. McCully 1
1Division of Cardiothoracic Surgery, Beth Israel Deaconess Medical Center and Harvard Medical School, 2Department of Anesthesiology, Perioperative and Pain Medicine, Boston Children's Hospital and Department of Anesthesia, Harvard Medical School

A method for rapid isolation of mitochondria from mammalian tissue biopsies is described. Rat liver or skeletal muscle preparations were homogenized with a commercial tissue dissociator and mitochondria were isolated by differential filtration through nylon mesh filters. Mitochondrial isolation time is <30 min compared to 60 - 100 min using alternative methods.

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