Peptides from Phage Display Library Modulate Gene Expression in Mesenchymal Cells and Potentiate Osteogenesis in Unicortical Bone Defects

JoVE 2362 12/10/2010

Gary Balian¹, Gina Beck¹, Vedavathi Madhu¹, Robert Sikes², Quanjun Cui³, Haixiang Liang¹, Joshua Bush¹

¹Orthopaedics Research, University of Virginia, ²Biological Sciences, University of Delaware, ³Orthopaedic Surgery, University of Virginia

A phage display library was used to identify peptide sequences that target bone. The objective was to investigate the effect of these peptides on mesenchymal cell differentiation and to determine their effect on bone regeneration.

Generation of Bone Marrow Derived Murine Dendritic Cells for Use in 2-photon Imaging

JoVE 773 7/09/2008

Melanie P. Matheu¹, Debasish Sen¹, Michael D Cahalan ¹, Ian Parker ²

¹Department of Physiology and Biophysics, University of California, Irvine (UCI), ²Department of Neurobiology and Behaviour, University of California, Irvine (UCI)

Antigen presentation in secondary lymphoid organs by dendritic cells is crucial for the initiation of the T cell mediated adaptive immune response. Here we demonstrate the culture of bone marrow derived murine dendritic cells, activation, and labeling for 2-photon imaging.

Pseudofracture: An Acute Peripheral Tissue Trauma Model

JoVE 2074 4/18/2011

Sophie S. Darwiche¹, Philipp Kobbe², Roman Pfeifer², Lauryn Kohut¹, Hans-Christoph Pape², Timothy Billiar¹

¹Department of Surgery, University of Pittsburgh, ²Department of Orthopedic Surgery, University of Aachen Medical Center

Pseudofracture, a reproducible murine model of sterile musculoskeletal trauma, allows for evaluation of late term post-traumatic immune responses. This article describes the procedural execution of the model step by step, including the potential for experimental model combinations to permit study of multiple trauma.

Culture of myeloid dendritic cells from bone marrow precursors

JoVE 769 7/25/2008

Jeanette Boudreau^1,2, Sandeep Koshy^2,3, Derek Cummings², Yonghong Wan²

¹Medical Sciences Program, McMaster University, ²Centre for Gene Therapeutics, McMaster University, ³Department of Chemical Engineering, University of Waterloo

This video demonstrates the procedure for differentiating myeloid dendritic cells from mouse bone marrow. Isolation of mouse tibia and femur, and processing of bone marrow are demonstrated. Pictures demonstrating cell morphology before and after differentiation, and figures depicting cell phenotype and IL-12 production following maturation using CpG are shown.

The Preparation of Primary Hematopoietic Cell Cultures From Murine Bone Marrow for Electroporation

JoVE 1026 1/06/2009

Kelly Kroeger, Michelle Collins, Luis Ugozzoli

Gene Expression Division, Bio-Rad Laboratories, Inc

This procedure describes how to establish primary hematopoietic cell cultures from murine bone marrow and is followed by transfection using the Gene Pulser MXCell electroporation system.

Homing of Hematopoietic Cells to the Bone Marrow

JoVE 1104 3/18/2009

Rushdia Z. Yusuf, David T. Scadden

Center for Regenerative Medicine, MGH - Massachusetts General Hospital

This article describes a protocol used to study the homing of hematopoietic cells to their niches in the bone marrow.

Generation and Labeling of Murine Bone Marrow-derived Dendritic Cells with Qdot Nanocrystals for Tracking Studies

JoVE 2785 6/02/2011

Maria Muccioli*¹, Michelle Pate*², Omowaleola Omosebi², Fabian Benencia^1,2,3

¹Molecular and Cell Biology Program, Ohio University, ²Department of Biomedical Sciences, College of Osteopathic Medicine, Ohio University, ³Department of Biomedical Engineering, Russ College of Engineering and Technology, Ohio University

Dendritic cells uptake antigens and migrate towards immune organs to present processed antigens to T cells. Qdot nanocrystal labeling provides a long-lasting and stable fluorescent signal. This allows tracking of dendritic cells to different organs by fluorescent microscopy.

Isolation and Transplantation of Hematopoietic Stem Cells (HSCs)

JoVE 157 2/25/2007

Cristina Lo Celso, David Scadden

Center for Regenerative Medicine, Massachusetts General Hospital, Harvard Stem Cell Institute, Harvard Medical School

Longitudinal Evaluation of Mouse Hind Limb Bone Loss After Spinal Cord Injury using Novel, in vivo, Methodology

JoVE 3246 12/07/2011

Madonna M. McManus, Raymond J. Grill

Department of Integrative Biology and Pharmacology, University of Texas Health Science Center at Houston

A longitudinal examination of bone loss in the femurs and tibiae of adult mice was performed following spinal cord injury using sequential low-dose X-ray scans. Tibia bone loss was detected throughout the study, while bone loss in the femur was not detected until 40 days post injury.

Creating Rigidly Stabilized Fractures for Assessing Intramembranous Ossification, Distraction Osteogenesis, or Healing of Critical Sized Defects

JoVE 3552 4/11/2012

Yan-yiu Yu, Chelsea Bahney, Diane Hu, Ralph S. Marcucio, Theodore Miclau, III

Department of Orthopaedic Surgery, University of California, San Francisco

This article describes a method for stabilizing long bone fractures that is based on the application of modified Ilizarov external fixators ^1-3. After application of the fixators and creation of the bone injury, healing can be assessed, distraction osteogenesis can be performed, or non-union or critical sized defect can be created and used to study therapeutic interventions.