Broad Institute of MIT and Harvard

10 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

Quantification of Proteins Using Peptide Immunoaffinity Enrichment Coupled with Mass Spectrometry
Lei Zhao *1, Jeffrey R. Whiteaker *1, Matthew E. Pope 2, Eric Kuhn 3, Angela Jackson 4, N. Leigh Anderson 5, Terry W. Pearson 2, Steven A. Carr 3, Amanda G. Paulovich 1
1Clinical Research Division, Fred Hutchinson Cancer Research Center - FHCRC, 2Department of Biochemistry and Microbiology, University of Victoria, 3Broad Institute of MIT and Harvard, 4Genome BC Proteomics Centre, University of Victoria, 5Plasma Proteome Institute

Stable Isotope Standards and Capture by Anti-Peptide Antibodies (SISCAPA) couples affinity enrichment of peptides with stable isotope dilution mass spectrometry (MRM-MS) to provide quantitative measurement of peptides as surrogates for their respective proteins. Here we describe the protocol using magnetic particles in a partially automated format.

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Medicine

Unbiased Deep Sequencing of RNA Viruses from Clinical Samples
Christian B. Matranga 1, Adrianne Gladden-Young 1, James Qu 1, Sarah Winnicki 1, Dolo Nosamiefan 1, Joshua Z. Levin 1, Pardis C. Sabeti 1,2
1Broad Institute of MIT and Harvard, 2Harvard University

This protocol describes a rapid and broadly applicable method for unbiased RNA-sequencing of viral samples from human clinical isolates.

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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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Neuroscience

Derivation, Expansion, Cryopreservation and Characterization of Brain Microvascular Endothelial Cells from Human Induced Pluripotent Stem Cells
Sovannarath Pong 1,2,3, Paulo Lizano 1,2,3,4, Rakesh Karmacharya 1,3,4,5
1Center for Genomic Medicine, Massachusetts General Hospital, 2Department of Psychiatry, Beth Israel Deaconess Medical Center, 3Chemical Biology and Therapeutic Science Program, Broad Institute of MIT and Harvard, 4Department of Psychiatry, Harvard Medical School, 5Schizophrenia and Bipolar Disorder Program, McLean Hospital

This protocol details an adapted method to derive, expand, and cryopreserve brain microvascular endothelial cells obtained by differentiating human induced pluripotent stem cells, and to study blood brain barrier properties in an ex vivo model.

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Cancer Research

Dissociation of Human and Mouse Tumor Tissue Samples for Single-cell RNA Sequencing
Jacy Fang *1,2, Isabella Salinas *1,2, Sarah San Vicente *1,2, Caroline Zielinski *1,2, Moshe Sade-Feldman 1,2,3
1Krantz Family Center for Cancer Research, Department of Medicine, Massachusetts General Hospital, 2Broad Institute of MIT and Harvard, 3Department of Medicine, Harvard Medical School

This protocol outlines the procedure for rapidly dissociating human and mouse tumor samples for single-cell RNA sequencing.

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Neuroscience

IntelliSleepScorer, a Software Package with a Graphic User Interface for Mice Automated Sleep Stage Scoring
Ziyue Zhu 1, Lei A. Wang 1, Ryan Kern 2, Jen Q. Pan 1
1Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, 2Stanford University

We present a software package with a graphic user interface for researchers without coding experience to score sleep stages in mice with a simple download and operation.