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Children’s Hospital Los Angeles

3 ARTICLES PUBLISHED IN JoVE

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Cancer Research

Aqueous Humor as a Liquid Biopsy for Retinoblastoma: Clear Corneal Paracentesis and Genomic Analysis
Mary E. Kim 1,2, Liya Xu 1,2,3,11, Rishvanth K. Prabakar 4, Lishuang Shen 5, Chen-Ching Peng 1,3, Peter Kuhn 3,6,7,8, Xiaowu Gai 5,9, James Hicks 3,6,10, Jesse L. Berry 1,2,6,11
1The Vision Center, Children’s Hospital Los Angeles, 2USC Roski Eye Institute, Keck School of Medicine of the University of Southern California, 3Department of Biological Sciences, Dornsife College of Letters, Arts, and Sciences, University of Southern California, 4Department of Molecular and Computational Biology, University of Southern California, 5Center for Personalized Medicine, Department of Pathology and Laboratory Medicine, Children’s Hospital Los Angeles, 6Norris Comprehensive Cancer Center, Keck School of Medicine, University of Southern California, 7Department of Aerospace and Mechanical Engineering, Viterbi School of Engineering, University of Southern California, 8Department of Biomedical Engineering, Viterbi School of Engineering, University of Southern California, 9Department of Pathology and Laboratory Medicine, Keck School of Medicine of USC, 10Department of Biochemistry and Molecular Medicine, Keck School of Medicine, University of Southern California, 11The Saban Research Institute, Children’s Hospital Los Angeles

The aqueous humor is a high-yield liquid biopsy for retinoblastoma, intraocular cancer that cannot be biopsied in vivo due to the risk of extraocular spread. Herein, a method for safely extracting aqueous humor via clear corneal paracentesis and steps for genomic analysis to identify prognostic biomarkers are presented.

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Developmental Biology

Modeling Paracrine Noncanonical Wnt Signaling In Vitro
Omar Toubat 1, Jongkyu Choi 1, S. Ram Kumar 1,2,3
1Department of Surgery, Keck School of Medicine of USC, 2Department of Pediatrics, Keck School of Medicine of USC, 3Heart Institute, Children’s Hospital Los Angeles

The present study outlines a highly reproducible and tractable method to study paracrine noncanonical Wnt signaling events in vitro. This protocol was applied to evaluate the impact of paracrine Wnt5a signaling in murine neural crest cells and myoblasts.

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Biology

Obtaining High-Quality Cryosections of Whole Rabbit Eye
Erik A. Souverein 1, Aaron Nagiel 2,3,4, Ksenia Gnedeva 5,6
1Keck School of Medicine, University of Southern California, 2The Vision Center, Department of Surgery, Children’s Hospital Los Angeles, 3The Saban Research Institute, Children’s Hospital Los Angeles, 4USC Roski Eye Institute, Department of Ophthalmology, Keck School of Medicine, University of Southern California, 5Eli and Edythe Broad CIRM Center for Regenerative Medicine and Stem Cell Research, University of Southern California, 6Tina and Rick Caruso Department of Otolaryngology-Head and Neck Surgery, University of Southern California

This protocol describes a reliable method for obtaining high-quality cryosections of whole rabbit eyes. It details rabbit eye dissection, fixation, embedding, and sectioning procedures, which may be easily adapted for use in any study utilizing immunohistochemistry in larger eyes.

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