NIH

5 articles published in JoVE

• Cancer Research

  
  Co-Culture and Transduction of Murine Thymocytes on Delta-Like 4-Expressing Stromal Cells to Study Oncogenes in T-Cell Leukemia Gisele O. L. Rodrigues¹, WenQing Li¹, Sarah D. Cramer^1,2,3, Hila Y. Winer¹, Tu Chun Hsu^1,2,3, Timothy Gower⁴, Julie A. Hixon¹, Scott K. Durum^{1 1}Cytokines and Immunity Section, Cancer Innovation Laboratory, National Cancer Institute, National Institutes of Health, ²Comparative Biomedical Scientist Training Program, NIH, ³Department of Pathobiology and Diagnostic Investigation, Veterinary Diagnostic Laboratory, Michigan State University, ⁴NCI-Frederick Laboratory Animal Sciences Program This protocol describes the isolation of double-negative thymocytes from the mouse thymus followed by retroviral transduction and co-culture on the delta-like 4-expressing bone marrow stromal cell line co-culture system (OP9-DL4) for further functional analysis.

• Immunology and Infection

  
  An In Vitro Caseum Binding Assay that Predicts Drug Penetration in Tuberculosis Lesions Jansy P. Sarathy¹, Hsin-pin Ho Liang¹, Danielle Weiner², Jacqueline Gonzales², Laura E. Via², Véronique Dartois^{1 1}Public Health Research Institute Centre, New Jersey Medical School, Rutgers, ²Tuberculosis Research Section, Laboratory of Clinical Infectious Diseases, NIAID, NIH Here we describe a rapid equilibrium dialysis (RED) method to measure drug binding to caseum from pulmonary tuberculosis lesions and cavities. The protocol is also used with a foamy macrophage-derived matrix that is an effective surrogate to caseum.

• Neuroscience

  
  Fluorescence Activated Cell Sorting (FACS) and Gene Expression Analysis of Fos-expressing Neurons from Fresh and Frozen Rat Brain Tissue F. Javier Rubio*¹, Xuan Li*¹, Qing-Rong Liu¹, Raffaello Cimbro², Bruce T. Hope^{1 1}Behavioral Neuroscience Research Branch, Intramural Research Program, NIDA, NIH, DHHS, ²Division of Rheumatology, School of Medicine, Johns Hopkins University Here we present a Fluorescence Activated Cell Sorting (FACS) protocol to study molecular alterations in Fos-expressing neuronal ensembles from both fresh and frozen brain tissue. The use of frozen tissue allows FACS isolation of many brain areas over multiple sessions to maximize the use of valuable animal subjects.

• Immunology and Infection

  
  Isolation of Leukocytes from the Murine Tissues at the Maternal-Fetal Interface Marcia Arenas-Hernandez¹, Elly N. Sanchez-Rodriguez¹, Tara N. Mial¹, Sarah A. Robertson², Nardhy Gomez-Lopez^{1,3,4 1}Department of Obstetrics & Gynecology, Wayne State University School of Medicine, ²School of Paediatrics and Reproductive Health, Research Centre for Reproductive Health, the Robinson Research Institute, The University of Adelaide, ³Department of Immunology & Microbiology, Wayne State University School of Medicine, ⁴Perinatology Research Branch, NICHD/NIH/DHHS Described herein is a protocol to isolate and analyze the infiltrating leukocytes of tissues at the maternal-fetal interface (uterus, decidua, and placenta) of mice. This protocol maintains the integrity of most cell surface markers and yields enough viable cells for downstream applications including flow cytometry analysis.

• Immunology and Infection

  
  Isolation of Leukocytes from the Human Maternal-fetal Interface Yi Xu¹, Olesya Plazyo¹, Roberto Romero^1,2,3,4, Sonia S. Hassan^1,5, Nardhy Gomez-Lopez^{1,5,6 1}Perinatology Research Branch, NICHD/NIH/DHHS, ²Department of Obstetrics and Gynecology, University of Michigan, ³Department of Epidemiology and Biostatistics, Michigan State University, ⁴Department of Molecular Obstetrics and Genetics, Wayne State University, ⁵Department of Obstetrics and Gynecology, Wayne State University School of Medicine, ⁶Department of Immunology and Microbiology, Wayne State University School of Medicine Described herein is a protocol to isolate and further study the infiltrating leukocytes of the decidua basalis and decidua parietalis - the human maternal-fetal interface. This protocol maintains the integrity of cell surface markers and yields enough viable cells for downstream applications as proven by flow cytometry analysis.