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Texas A&M University Health Science Center

9 ARTICLES PUBLISHED IN JoVE

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Biology

Microfluidic Co-culture of Epithelial Cells and Bacteria for Investigating Soluble Signal-mediated Interactions
Jeongyun Kim 1, Manjunath Hegde 1, Arul Jayaraman 1,2
1McFerrin Department of Chemical Engineering, Texas A&M University, 2Department of Biomedical Engineering, Texas A&M University

This protocol describes a microfluidic co-culture model for simultaneous and localized culture of epithelial cells and bacteria. This model can be used for investigating the role of different soluble molecular signals on pathogenesis as well as screen the effectiveness of putative probiotic bacterial strains.

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Biology

A Microfluidic Device for Quantifying Bacterial Chemotaxis in Stable Concentration Gradients
Derek L. Englert 1, Michael D. Manson 2, Arul Jayaraman 1,3
1McFerrin Department of Chemical Engineering, Texas A&M University, 2Department of Biology, Texas A&M University, 3Department of Biomedical Engineering, Texas A&M University

This protocol describes the development of a microfluidic device for investigating bacterial chemotaxis in stable concentration gradients of chemoeffectors.

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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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Developmental Biology

Production and Administration of Therapeutic Mesenchymal Stem/Stromal Cell (MSC) Spheroids Primed in 3-D Cultures Under Xeno-free Conditions
Joni H. Ylostalo 1, Nikolay Bazhanov 2, Arezoo Mohammadipoor 3, Thomas J. Bartosh 4
1Department of Biology, University of Mary Hardin-Baylor, 2Department of Pediatrics, University of Texas Medical Branch, 3Multi-Organ Support Technology Task Area, U.S. Army Institute of Surgical Research, 4Internal Medicine, Texas A&M University Health Science Center

The therapeutic potential of mesenchymal stem/stromal cells (MSCs) is well-documented, however the best method of preparing the cells for patients remains controversial. Herein, we communicate protocols to efficiently generate and administer therapeutic spherical aggregates or 'spheroids' of MSCs primed under xeno-free conditions for experimental and clinical applications.

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Immunology and Infection

Examination of Host Phenotypes in Gambusia affinis Following Antibiotic Treatment
Jeanette M. Carlson 1, Oscar Chavez 1,2, Sonali Aggarwal 1, Todd P. Primm 1
1Biological Sciences, Sam Houston State University, 2Irma Lerma Rangel College of Pharmacy, Texas A&M University Health Science Center

This study involves methods to reveal effects on a model fish host following alteration of the skin and gut microbiome communities composition by an antibiotic.

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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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Immunology and Infection

A Comprehensive High-Efficiency Protocol for Isolation, Culture, Polarization, and Glycolytic Characterization of Bone Marrow-Derived Macrophages
Sahar Eshghjoo 1, Da Mi Kim 2, Arul Jayaraman 1,3, Yuxiang Sun 2, Robert C. Alaniz 1
1Department of Microbial Pathogenesis and Immunology, College of Medicine, Texas A&M University Health Science Center, 2Department of Nutrition and Food Science, Texas A&M University, 3Artie McFerrin Department of Chemical Engineering, Texas A&M University

This protocol provides detailed and comprehensive methods for the isolation, culture, polarization, and measurement of the glycolytic metabolic state of live bone marrow-derived macrophages (BMDMs). This paper provides step-by-step instructions with realistic visual illustrations for workflow and glycolytic assessment of BMDMs in real-time.

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Cancer Research

Efficient Gene Knockdown in the Liver via Intrasplenic Injection of Adeno-Associated Virus Serotype 8 (AAV8)-Delivered Small Hairpin RNA
Xiaoqin Liu 1, Robert Y. L. Tsai 1,2
1Institute of Biosciences and Technology, Texas A&M University Health Science Center, 2Department of Translational Medical Sciences, College of Medicine, Texas A&M University Health Science Center

This protocol describes how intrasplenic injection of AAV8-delivered small hairpin RNA achieves the same gene knockdown efficiency in the liver as portal vein injection, representing a simpler procedure with much lower perioperative and postoperative mortality and complications.

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