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The Pennsylvania State University College of Medicine

8 ARTICLES PUBLISHED IN JoVE

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Immunology and Infection

Directed Differentiation of Induced Pluripotent Stem Cells towards T Lymphocytes
Fengyang Lei 1, Rizwanul Haque 1, Xiaofang Xiong 1, Jianxun Song 1
1Department of Microbiology and Immunology, Pennsylvania State University College of Medicine

Generation of T lymphocytes from induced pluripotent stem (iPS) cells gives an alternative approach of using embryonic stem cells for T cell-based immunotherapy. The method shows that by utilizing either in vitro or in vivo induction system, iPS cells are able to differentiate into both conventional and antigen-specific T lymphocytes.

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Immunology and Infection

Development of Stem Cell-derived Antigen-specific Regulatory T Cells Against Autoimmunity
Mohammad Haque 1, Kristin Fino 1, Praneet Sandhu 1, Jianxun Song 1
1Department of Microbiology and Immunology, The Pennsylvania State University College of Medicine

We present here a method to develop functional antigen (Ag)-specific regulatory T cells (Tregs) from induced pluripotent stem cells (iPSCs) for immunotherapy of autoimmune arthritis in a murine model.

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Immunology and Infection

Stem Cell-Derived Viral Ag-Specific T Lymphocytes Suppress HBV Replication in Mice
Xiaofang Xiong 1, Fengyang Lei 2, Mohammad Haque 1, Jianxun Song 1
1Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, 2Department of Ophthalmology, Harvard Medical School

Presented here is a protocol for the effective suppression of hepatitis B virus (HBV) replication in mice by utilizing adoptive cell transfer (ACT) of stem cell-derived viral antigen (Ag)-specific T lymphocytes. This procedure may be adapted for potential ACT-based immunotherapy of HBV infection.

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Medicine

Standardized Histomorphometric Evaluation of Osteoarthritis in a Surgical Mouse Model
William J. Pinamont 1, Natalie K. Yoshioka 1, Gregory M. Young 1, Vengadeshprabhu Karuppagounder 1, Elijah L. Carlson 1, Adeel Ahmad 1, Reyad Elbarbary 1,2, Fadia Kamal 1,3
1Center for Orthopedic Research and Translational Sciences, Department of Orthopedics and Rehabilitation, Pennsylvania State College of Medicine, 2Department of Biochemistry and Molecular Biology, Pennsylvania State College of Medicine, 3Department of Pharmacology, Pennsylvania State College of Medicine

The current protocol establishes a rigorous and reproducible method for quantification of morphological joint changes that accompany osteoarthritis. Application of this protocol can be valuable in monitoring disease progression and evaluating therapeutic interventions in osteoarthritis.

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Neuroscience

Traumatic Peripheral Nerve Injury in Mice
Jung Il Lee *1, Prem Kumar Govindappa *2, Grant D. Wandling 2, John C. Elfar 2
1Department of Orthopaedic Surgery, Korea University Guro Hospital, 2Department of Orthopaedics and Rehabilitation, Center for Orthopaedics and Translational Science, The Pennsylvania State University College of Medicine

The present protocol describes traumatic peripheral nerve injuries (TPNIs), including precisely calibrated crush, strictly aligned and misaligned laceration, as well as grafted and non-grafted gaps of the sciatic nerve in mice. Custom-designed sensors are developed to gauge nerve trauma, induced with commonly available tools to ensure reproducible post-TPNI outcomes.

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Neuroscience

Isolation, Culture, and Characterization of Primary Schwann Cells, Keratinocytes, and Fibroblasts from Human Foreskin
Mashanipalya G. Jagadeeshaprasad 1, Prem Kumar Govindappa 1, Amanda M. Nelson 2, John C. Elfar 1
1Department of Orthopaedics and Rehabilitation, Center for Orthopaedic Research and Translational Science (CORTS), The Pennsylvania State University College of Medicine, 2Department of Dermatology, The Pennsylvania State University College of Medicine

The study of wound healing associated with musculoskeletal injury often requires the assessment of in vitro interactions between Schwann cells (SCs), keratinocytes, and fibroblasts. This protocol describes the isolation, culturing, and characterization of these primary cells from the human foreskin.

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Biology

Assessment of Bone Fracture Healing Using Micro-Computed Tomography
Hwabok Wee *1,2, Deepak Kumar Khajuria *1,2, Fadia Kamal 1,2,3, Gregory S. Lewis 1,2, Reyad A. Elbarbary 1,2,4
1Center for Orthopaedic Research and Translational Science (CORTS), The Pennsylvania State University College of Medicine, 2Department of Orthopaedics and Rehabilitation, The Pennsylvania State University College of Medicine, 3Department of Pharmacology, The Pennsylvania State University College of Medicine, 4Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine

Micro-computed tomography (µCT) is a non-destructive imaging tool that is instrumental in assessing bone structure in preclinical studies, however there is a lack of consensus on µCT procedures for analyzing the bone healing callus. This study provides a step-by-step µCT protocol that allows the monitoring of fracture healing.

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Musculoskeletal Research: Current Methods and Innovative Approaches
Reyad A. Elbarbary 1,2,3,4
1Center for Orthopaedic Research and Translational Science (CORTS), The Pennsylvania State University College of Medicine, 2Department of Orthopaedics and Rehabilitation, The Pennsylvania State University College of Medicine, 3Department of Biochemistry and Molecular Biology, The Pennsylvania State University College of Medicine, 4Center for RNA Molecular Biology, Pennsylvania State University

Musculoskeletal Research: Current Methods and Innovative Approaches

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