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11 ARTICLES PUBLISHED IN JoVE

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Biology

Passaging HuES Human Embryonic Stem Cell-lines with Trypsin.
Erin Trish 1, John Dimos 1, Kevin Eggan 1
1Department of Molecular and Cell Biology, Harvard

In this video we demonstrate how our lab routinely passages HuES human embryonic stem cell lines with trypsin. Brought to you by JoVE.

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Biology

Freezing Human ES Cells
Erin Trish 1, John Dimos 1, Kevin Eggan 1
1Department of Molecular and Cell Biology, Harvard

Here we demonstrate how our lab freezes HuES human embryonic stem cell lines.

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Biology

Human ES cells: Starting Culture from Frozen Cells
Erin Trish 1, John Dimos 1, Kevin Eggan 1
1Department of Molecular and Cell Biology, Harvard

Here we demonstrate how our lab begins a HuES human embryonic stem cell line culture from a frozen stock.

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Biology

Nuclear Transfer into Mouse Oocytes
Dieter Egli 1, Kevin Eggan 1
1Department of Molecular and Cell Biology, Harvard

This movie and the protocol are intended to help learning nuclear transfer.

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Biology

Measuring Exocytosis in Neurons Using FM Labeling
Jamila Newton 1, Venkatesh Murthy 1
1Department of Molecular and Cell Biology, Harvard

The ability to measure the kinetics of vesicle release can help provide insight into some of the basics of neurotransmission. Here we used real-time imaging of vesicles labeled with the red fluorescent dye FM 4-64 to measure the rate of presynaptic vesicle release in hippocampal neuronal cultures.

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Biology

Derivation of Human Embryonic Stem Cells by Immunosurgery
Alice E. Chen 1, Douglas A. Melton 1
1Department of Molecular and Cell Biology, Harvard

The ability of human embryonic stem cells to self-renew and differentiate into all cell types of the body suggests that they hold great promise for both medical applications and as a research tool for addressing fundamental questions in development and disease. Here, we provide a concise, step-by-step protocol for the derivation of human embryonic stem cells from embryos by immunosurgical isolation of the inner cell mass.

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Biology

Studying Membrane Biogenesis with a Luciferase-Based Reporter Gene Assay
Shaochong Zhang 1, Axel Nohturfft 1,2
1Department of Molecular and Cell Biology, Harvard, 2Molecular and Metabolic Signalling Centre, Division of Basic Medical Sciences, St. George's University of London

Here, we describe procedures for studying changes in phagocytosis-induced gene expression with a luciferase-based reporter gene approach using the Dual-GloTM Luciferase Assay System from Promega.

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Biology

A Behavioral Assay to Measure Responsiveness of Zebrafish to Changes in Light Intensities
Farida Emran 1, Jason Rihel 1, John E. Dowling 1
1Department of Molecular and Cell Biology, Harvard

We developed the Visual-Motor Response to quantitate the motor output of larval zebrafish in response to light increments and decrements. We also examined zebrafish vision mutants, including the no optokinetic response (nrc) mutants, which were thought to be completely blind when tested by another vision assay, the optokinetic reflex.

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Biology

Making MR Imaging Child's Play - Pediatric Neuroimaging Protocol, Guidelines and Procedure
Nora M. Raschle 1,2, Michelle Lee 1, Roman Buechler 1, Joanna A. Christodoulou 3, Maria Chang 1, Monica Vakil 1, Patrice L. Stering 1, Nadine Gaab 1,3,4
1Department of Developmental Medicine, Children’s Hospital Boston, 2Department of Neuropsychology, University of Zurich, 3Graduate School of Education, Harvard, 4Harvard Medical School

Despite an increase in the use of structural and functional magnetic resonance imaging (fMRI) in humans, the study of young pediatric populations remains a challenge. We present a hands-on, step-by-step video protocol including guidelines for clinicians and researchers intending to perform (f)MRI in young children.

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Biology

Primary Culture and Plasmid Electroporation of the Murine Organ of Corti.
Mark Parker 1,2,3, Aurore Brugeaud 1,2, Albert S. B. Edge 1,2,4
1Department of Otology and Laryngology, Harvard Medical School, 2Eaton-Peabody Laboratory, Massachusetts Eye and Ear Infirmary, 3Department of Communication Sciences and Disorders, Emerson College, 4Program in Speech and Hearing Bioscience and Technology, Division of Health Science and Technology, Harvard

This procedure describes a method for the isolation and culture of the murine organ of Corti with or without the spiral limbus and spiral ganglion neurons. We also demonstrate a method for the expression of an exogenous reporter gene in the organ of Corti explant by electroporation.

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Neuroscience

Modelling Zika Virus Infection of the Developing Human Brain In Vitro Using Stem Cell Derived Cerebral Organoids
Max R Salick *1, Michael F Wells *2,3, Kevin Eggan 2,3, Ajamete Kaykas 1
1Department of Neuroscience, Novartis Institutes for BioMedical Research, 2Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, 3Department of Stem Cell and Regenerative Biology and Harvard Stem Cell Institute, Harvard University

This protocol describes a technique used to model Zika virus infection of the developing human brain. Using wildtype or engineered stem cell lines, researchers may use this technique to uncover the various mechanisms or treatments that may affect early brain infection and resulting microcephaly in Zika virus-infected embryos.

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