The George Washington University School of Medicine and Health Sciences

2 ARTICLES PUBLISHED IN JoVE

Neuroscience

Isolation of Cerebrospinal Fluid from Rodent Embryos for use with Dissected Cerebral Cortical Explants

Mauro W. Zappaterra¹, Anthony S. LaMantia², Christopher A. Walsh^3,4, Maria K. Lehtinen⁵

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.

Cancer Research

Detection and Monitoring of Tumor Associated Circulating DNA in Patient Biofluids

Erin R. Bonner^1,2, Karim Saoud¹, Sulgi Lee^1,2, Eshini Panditharatna³, Madhuri Kambhampati¹, 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.