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Pennsylvania State University College of Medicine

9 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

Accelerated Type 1 Diabetes Induction in Mice by Adoptive Transfer of Diabetogenic CD4+ T Cells
Gregory Berry 1, Hanspeter Waldner 1
1Department of Microbiology & Immunology, Pennsylvania State University College of Medicine

We provide a reproducible method to induce type 1 diabetes (T1D) in mice within two weeks by the adoptive transfer of islet antigen-specific, primary CD4+ T cells.

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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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Neuroscience

A Free-breathing fMRI Method to Study Human Olfactory Function
Jianli Wang 1, Sebastian Rupprecht 1, Xiaoyu Sun 1, Diana Freiberg 1, Courtney Crowell 1, Emma Cartisano 1, Megha Vasavada 1, Qing X. Yang 1,2
1Center for NMR Research, Department of Radiology, Pennsylvania State University College of Medicine, 2Department of Neurosurgery, Pennsylvania State University College of Medicine

We present the technical challenges and solutions for obtaining reliable functional magnetic resonance imaging (fMRI) data from the human central olfactory system. This includes special considerations in olfactory fMRI paradigm design, descriptions of fMRI data acquisition with an MRI-compatible olfactometer, odorant selection, and a special software tool for data post-processing.

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Immunology and Infection

Lung microRNA Profiling Across the Estrous Cycle in Ozone-exposed Mice
Nathalie Fuentes 1, Patricia Silveyra 1,2
1Pulmonary, Immunology and Physiology Laboratory, Department of Pediatrics, Pennsylvania State University College of Medicine, 2Department of Biochemistry and Molecular Biology, Pennsylvania State University College of Medicine

Here we describe a method to assess lung expression of miRNAs that are predicted to regulate inflammatory genes using mice exposed to ozone or filtered air at different stages of the estrous cycle.

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Genetics

HOX Loci Focused CRISPR/sgRNA Library Screening Identifying Critical CTCF Boundaries
Huacheng Luo 1, Amin Sobh 2, Christopher D. Vulpe 2, Edmond Brewer 1, Sinisa Dovat 1, Yi Qiu 3, Suming Huang 1
1Department of Pediatrics, Pennsylvania State University College of Medicine, 2Department of Physiological Sciences, University of Florida, 3Department of Anatomy and Cell Biology, University of Florida

A CRISPR/sgRNA library has been applied to interrogating protein-coding genes.However, the feasibility of a sgRNA library to uncover the function of a CTCF boundary in gene regulation remains unexplored. Here, we describe a HOX loci specific sgRNA library to elucidate the function of CTCF boundaries in HOX loci.

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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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A Xenograft Mouse Model to Assess Efficacy of Therapeutic Agents for Human Acute Leukemia
Chandrika Gowda *1, Charyguly Annageldiyev *2,3, Pavan Kumar Dhanyamraju 1, Morgann Klink 1, Sinisa Dovat 1, Mark Kester 4,5, Thomas P. Loughran, Jr 4,6, David Claxton 2,3, Arati Sharma 1,2,7
1Department of Pediatrics, Pennsylvania State University College of Medicine, 2Penn State Hershey Cancer Institute, Pennsylvania State University College of Medicine, 3Departments of Medicine, Division of Hematology and Oncology, Pennsylvania State University College of Medicine, 4University of Virginia Cancer Center, 5nanoSTAR Institute, University of Virginia, 6Division of Hematology and Oncology, Department of Medicine, University of Virginia School of Medicine, 7Department of Pharmacology, Pennsylvania State University College of Medicine

Mouse (Mus Musculus) models are being widely used to develop xenografts using human leukemia cells. These models provide a comparable biological system to study drug efficacy, pharmacodynamics, and pharmacokinetics. Modeling acute myeloid leukemia in immunocompromised mice is described in detail using the U937 cell line xenograft as an example.

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Biology

3D Culturing of Organoids from the Intestinal Villi Epithelium Undergoing Dedifferentiation
Christina Li 1, Jeel Shah 1, Kylee Wrath 1, Dahlia Matouba 1, Connor Mills 1, Kishore Punnath 2, Ansu Perekatt 1
1Department of Biology and Chemical Biology, Stevens Institute of Technology, 2Department of Pharmacology, Pennsylvania State University College of Medicine

The procedure describes isolation of the villi from the mouse intestinal epithelium undergoing dedifferentiation to determine their organoid forming potential.

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