JoVE Monthly Highlights: October 2018

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1JoVE Content Production

Immunophenotyping of Orthotopic Homograft (Syngeneic) of Murine Primary KPC Pancreatic Ductal Adenocarcinoma by Flow Cytometry

Xiaoyu An*1,2, Xuesong Ouyang*1, Hui Zhang1, Tingting Li1, Yu-yang Huang1, Zhiyuan Li1, Demin Zhou2, Qi-Xang Li1,2
1Crown Bioscience Inc., 2State Key Laboratory of Natural and Biomimetic Drugs, Peking University
* These authors contributed equally

The experimental procedure on the immunophenotyping of murine orthotopic PDAC homografts aims at profiling the tumor immuno-microenvironment. Tumors are orthotopically implanted via surgery. Tumors of 200–600 mm3 in size were harvested and dissociated to prepare single-cell suspensions, followed by multi-immune marker FACS analysis using different fluorescently-labeled antibodies.

Fabrication and Testing of Photonic Thermometers

Nikolai N. Klimov1,2, Zeeshan Ahmed2
1Joint Quantum Institute, University of Maryland, 2Physical Measurement Laboratory, National Institute of Standards and Technology

We describe the process of fabrication and testing of photonic thermometers.

Intravenous and Intra-amniotic In Utero Transplantation in the Murine Model

Nicholas J. Ahn1, John D. Stratigis1, Barbara E. Coons1, Alan W. Flake1, Hyun-Duck Nah-Cederquist2, William H. Peranteau1
1Division of General, Thoracic, and Fetal Surgery, Center for Fetal Research, Children's Hospital of Philadelphia, 2Division of Plastic Reconstructive Surgery, Center for Fetal Research, Children's Hospital of Philadelphia

We describe a protocol for performing an in utero transplantation (IUT) through intravenous and intra-amniotic routes of injection in the murine model. This protocol can be used to introduce cells, viral vectors, and other substances into the unique immune-tolerant fetal environment.

A Rat Methyl-Seq Platform to Identify Epigenetic Changes Associated with Stress Exposure

Jenny L. Carey1, Olivia H. Cox1, Fayaz Seifuddin1, Leonard Marque1, Kellie L.K. Tamashiro1, Peter P. Zandi1,3, Gary S. Wand1,2, Richard S. Lee1
1Departments of Psychiatry and Behavioral Sciences, Johns Hopkins School of Medicine, 2Department of Medicine, Johns Hopkins School of Medicine, 3Department of Mental Health, Johns Hopkins School of Public Health

Here, we describe the protocol and implementation of Methyl-Seq, an epigenomic platform, using a rat model to identify epigenetic changes associated with chronic stress exposure. Results demonstrate that the rat Methyl-Seq platform is capable of detecting methylation differences that arise from stress exposure in rats.

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