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Boston Children's Hospital and Department of Anesthesia, Harvard Medical School

5 ARTICLES PUBLISHED IN JoVE

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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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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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