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The George Washington University School of Medicine and Health Sciences

2 ARTICLES PUBLISHED IN JoVE

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Neuroscience

Isolation of Cerebrospinal Fluid from Rodent Embryos for use with Dissected Cerebral Cortical Explants
Mauro W. Zappaterra 1, Anthony S. LaMantia 2, Christopher A. Walsh 3,4, Maria K. Lehtinen 5
1Department of Physical Medicine and Rehabilitation, VA Greater Los Angeles Healthcare System, 2Department of Pharmacology and Physiology, Institute for Neuroscience, The George Washington University School of Medicine and Health Sciences, 3Division of Genetics, Department of Medicine, Boston Children's Hospital, 4Howard Hughes Medical Institute, Boston Children's Hospital, 5Department of Pathology, Boston Children's Hospital, Harvard Medical School

The ventricular cerebrospinal fluid (CSF) bathes the neuroepithelial and cerebral cortical progenitor cells during early brain development in the embryo. Here we describe the method developed to isolate ventricular CSF from rodent embryos of different ages in order to investigate its biological function. In addition, we demonstrate our cerebral cortical explant dissection and culture technique that allows for explant growth with minimal volumes of culture medium or CSF.

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Cancer Research

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids
Erin R. Bonner 1,2, Karim Saoud 1, Sulgi Lee 1,2, Eshini Panditharatna 3, Madhuri Kambhampati 1, Sabine Mueller 4,5, Javad Nazarian 1,2,5
1Center for Genetic Medicine, Children's National Health System, 2Institute for Biomedical Sciences, The George Washington University School of Medicine and Health Sciences, 3Dana-Farber Cancer Institute, 4Department of Neurology, University of California San Francisco, 5DIPG Centre of Expertise Zurich, Universitats-Kinderspital Zurich

Here, we present a protocol to detect tumor somatic mutations in circulating DNA present in patient biological fluids (biofluids). Our droplet digital polymerase chain reaction (dPCR)-based method enables quantification of the tumor mutation allelic frequency (MAF), facilitating a minimally invasive complement to diagnosis and temporal monitoring of tumor response.

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