University of Texas M.D. Anderson Cancer Center

8 ARTICLES PUBLISHED IN JoVE

Biology

Selecting and Isolating Colonies of Human Induced Pluripotent Stem Cells Reprogrammed from Adult Fibroblasts

Urszula Polak^1,2, Calley Hirsch¹, Sherman Ku³, Joel Gottesfeld³, Sharon Y.R. Dent¹, Marek Napierala¹

¹Department of Molecular Carcinogenesis and Center for Cancer Epigenetics, University of Texas M.D. Anderson Cancer Center, ²Department of Cell Biology, Poznan University of Medical Sciences, ³Department of Molecular Biology, The Scripps Research Institute

We present a protocol for efficient reprogramming of human somatic cells into human induced pluripotent stem cells (hiPSC) using retroviral vectors encoding Oct3/4, Sox2, Klf4 and c-myc (OSKM) and identification of correctly reprogrammed hiPSC by live staining with Tra-1-81 antibody.

Immunology and Infection

Expanding Cytotoxic T Lymphocytes from Umbilical Cord Blood that Target Cytomegalovirus, Epstein-Barr Virus, and Adenovirus

Patrick J. Hanley¹, Sharon Lam^1,2, Elizabeth J. Shpall³, Catherine M. Bollard^1,2,4,5

¹Center for Cell and Gene Therapy, Baylor College of Medicine , ²Pathology and Immunology, Baylor College of Medicine , ³Department of Stem Cell Transplantation and Cellular Therapy, University of Texas M.D. Anderson Cancer Center, ⁴Medicine, Baylor College of Medicine , ⁵Department of Pediatrics, Baylor College of Medicine

Here we describe the first good manufacturing practice (GMP)-compliant method of producing virus-specific cytotoxic T lymphocytes (CTL) from umbilical cord blood, a source of predominantly naîve T cells.

Immunology and Infection

Clinical Application of Sleeping Beauty and Artificial Antigen Presenting Cells to Genetically Modify T Cells from Peripheral and Umbilical Cord Blood

M. Helen Huls¹, Matthew J. Figliola¹, Margaret J. Dawson¹, Simon Olivares¹, Partow Kebriaei², Elizabeth J. Shpall², Richard E. Champlin², Harjeet Singh¹, Laurence J.N. Cooper¹

¹Division of Pediatrics, U.T. MD Anderson Cancer Center, ²Department of Stem Cell Transplantation and Cellular Therapy, U.T. MD Anderson Cancer Center

T cells expressing a CD19-specific chimeric antigen receptor (CAR) are infused as investigational treatment of B-cell malignancies in our first-in-human gene therapy trials. We describe genetic modification of T cells using the Sleeping Beauty (SB) system to introduce CD19-specific CAR and selective propagation on designer CD19+ artificial antigen presenting cells.

JoVE Journal

Protocols for Assessing Radiofrequency Interactions with Gold Nanoparticles and Biological Systems for Non-invasive Hyperthermia Cancer Therapy

Stuart J. Corr^1,2, Brandon T. Cisneros^1,2, Leila Green¹, Mustafa Raoof¹, Steven A. Curley^1,3

¹Department of Surgical Oncology, University of Texas M.D. Anderson Cancer Center, ²Department of Chemistry, Rice University , ³Mechanical Engineering and Materials Science, Rice University

We describe the protocols used to investigate the interactions of 13.56 MHz radiofrequency (RF) electric-fields with gold nanoparticle colloids in both non-biological and biological systems (in vitro/vivo). These interactions are being investigated for applications in cancer therapy.

Chemistry

Synthesis of Immunotargeted Magneto-plasmonic Nanoclusters

Chun-Hsien Wu^1,2, Konstantin Sokolov^1,2

¹Department of Biomedical Engineering, University of Texas at Austin, ²Department of Imaging Physics, University of Texas M.D. Anderson Cancer Center

Here, we describe a protocol for synthesis of magneto-plasmonic nanoparticles with a strong magnetic moment and a strong near-infrared (NIR) absorbance. The protocol also includes antibody conjugation to the nanoparticles through the Fc moiety for various biomedical applications which require molecular specific targeting.

Immunology and Infection

Automated Cell Enrichment of Cytomegalovirus-specific T cells for Clinical Applications using the Cytokine-capture System

Pappanaicken Kumaresan^*1, Mathew Figliola^*1, Judy S. Moyes¹, M. Helen Huls¹, Priti Tewari¹, Elizabeth J. Shpall², Richard Champlin², Laurence J.N. Cooper¹

¹Division of Pediatrics, U.T. MD Anderson Cancer Center, ²Stem Cell Transplantation and Cellular Therapy, U.T. MD Anderson Cancer Center

The goal of this protocol is to manufacture pathogen-specific clinical-grade T cells using a bench-top, automated, second generation cell enrichment device that incorporates a closed cytokine capture system and does not require dedicated staff or use of a GMP facility. The cytomegalovirus pp65-specific-T cells generated can be directly administered to patients.

Engineering

Use of a Multi-compartment Dynamic Single Enzyme Phantom for Studies of Hyperpolarized Magnetic Resonance Agents

Christopher M. Walker¹, Matthew Merritt², Jian-Xiong Wang², James A. Bankson¹

¹Imaging Physics, University of Texas M.D. Anderson Cancer Center, ²Advanced Imaging Research Center, University of Texas Southwestern Medical Center

A multi-compartment dynamic phantom is used to simulate some biology of interest for metabolic studies using hyperpolarized magnet resonance agents.

Medicine

Combined Near-infrared Fluorescent Imaging and Micro-computed Tomography for Directly Visualizing Cerebral Thromboemboli

Dong-Eog Kim¹, Jeong-Yeon Kim¹, Su-Kyoung Lee¹, Ju Hee Ryu², Ick Chan kwon², Cheol-Hee Ahn³, Kwangmeyung Kim², Dawid Schellingerhout⁴

¹Molecular Imaging and Neurovascular Research Laboratory, Dongguk University College of Medicine, ²Biomedical Research Center, Korea Institute of Science and Technology, ³Research Institute of Advanced Materials, Department of Materials Science and Engineering, Seoul National University, ⁴Departments of Radiology and Cancer Systems Imaging, University of Texas M.D. Anderson Cancer Center

This protocol describes the application of combined near-infrared fluorescent (NIRF) imaging and micro-computed tomography (microCT) for visualizing cerebral thromboemboli. This technique allows the quantification of thrombus burden and evolution. The NIRF imaging technique visualizes fluorescently labeled thrombus in excised brain, while the microCT technique visualizes thrombus inside living animals using gold-nanoparticles.