Generation of cortical neurons from mouse embryonic stem cells.

Abstract:

:Embryonic stem cells (ESCs) constitute a tool of great potential in neurobiology, enabling the directed differentiation of specific neural cell types. We have shown recently that neurons of the cerebral cortex can be generated from mouse ESCs cultured in a chemically defined medium that contains no morphogen, but in the presence of the sonic hedgehog inhibitor cyclopamine. Corticogenesis from ESCs recapitulates the most important steps of cortical development, leading to the generation of multipotent cortical progenitors that sequentially produce cortical pyramidal neurons displaying distinct layer-specific identities. The protocol provides a most reductionist cellular model to tackle the complex mechanisms of cortical development and function, thereby opening new perspectives for the modeling of cortical diseases and the design of novel neurological treatments, while offering an alternative to animal use. In this protocol, we describe a method by which millions of cortical neurons can be generated in 2-3 weeks, starting from a single frozen vial of ESCs.

journal_name

Nat Protoc

journal_title

Nature protocols

authors

Gaspard N,Bouschet T,Herpoel A,Naeije G,van den Ameele J,Vanderhaeghen P

doi

10.1038/nprot.2009.157

subject

Has Abstract

pub_date

2009-01-01 00:00:00

pages

1454-63

issue

10

eissn

1754-2189

issn

1750-2799

pii

nprot.2009.157

journal_volume

4

pub_type

杂志文章
  • Determining the polysaccharide composition of plant cell walls.

    abstract::The plant cell wall is a chemically complex structure composed mostly of polysaccharides. Detailed analyses of these cell wall polysaccharides are essential for our understanding of plant development and for our use of plant biomass (largely wall material) in the food, agriculture, fabric, timber, biofuel and biocompo...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2012.081

    authors: Pettolino FA,Walsh C,Fincher GB,Bacic A

    更新日期:2012-09-01 00:00:00

  • Nucleotide resolution profiling of m7G tRNA modification by TRAC-Seq.

    abstract::Precise identification of sites of RNA modification is key to studying the functional role of such modifications in the regulation of gene expression and for elucidating relevance to diverse physiological processes. tRNA reduction and cleavage sequencing (TRAC-Seq) is a chemically based approach for the unbiased globa...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-019-0226-7

    authors: Lin S,Liu Q,Jiang YZ,Gregory RI

    更新日期:2019-11-01 00:00:00

  • Methods for isolation, purification and structural elucidation of bioactive secondary metabolites from marine invertebrates.

    abstract::In the past few decades, marine natural products bioprospecting has yielded a considerable number of drug candidates. Two marine natural products have recently been admitted as new drugs: Prialt (also known as ziconotide) as a potent analgesic for severe chronic pain and Yondelis (known also as trabectedin or E-743) a...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2008.182

    authors: Ebada SS,Edrada RA,Lin W,Proksch P

    更新日期:2008-01-01 00:00:00

  • Polyethylene glycol-based bidentate ligands to enhance quantum dot and gold nanoparticle stability in biological media.

    abstract::We describe a simple and versatile scheme to prepare a series of poly(ethylene glycol)-based bidentate ligands that permit strong interactions with colloidal semiconductor nanocrystals (quantum dots, QDs) and gold nanoparticles (AuNPs) alike and promote their dispersion in aqueous solutions. These ligands are synthesi...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2008.243

    authors: Mei BC,Susumu K,Medintz IL,Mattoussi H

    更新日期:2009-01-01 00:00:00

  • Preparation, functionalization and characterization of engineered carbon nanodots.

    abstract::Carbon-based dots (CDs) and their functionalized (nano)composites have recently attracted attention due to their seemingly easy preparation and numerous potential applications, ranging from those in the biomedical field (i.e., imaging and drug delivery) to those in (opto)electronics (i.e., solar cells and LEDs). This ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-019-0207-x

    authors: Ðorđević L,Arcudi F,Prato M

    更新日期:2019-10-01 00:00:00

  • Population-specific genotype imputations using minimac or IMPUTE2.

    abstract::In order to meaningfully analyze common and rare genetic variants, results from genome-wide association studies (GWASs) of multiple cohorts need to be combined in a meta-analysis in order to obtain enough power. This requires all cohorts to have the same single-nucleotide polymorphisms (SNPs) in their GWASs. To this e...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2015.077

    authors: van Leeuwen EM,Kanterakis A,Deelen P,Kattenberg MV,Genome of the Netherlands Consortium.,Slagboom PE,de Bakker PI,Wijmenga C,Swertz MA,Boomsma DI,van Duijn CM,Karssen LC,Hottenga JJ

    更新日期:2015-09-01 00:00:00

  • Application of cationic conjugated polymers in microarrays using label-free DNA targets.

    abstract::A fluorescence-based microarray technique that does not require target DNA labeling is detailed. This 'label-free' approach utilizes a cationic, water-soluble conjugated polymer PFBT (poly[9,9'-bis(6''-(N,N,N-trimethylammonium)hexyl)fluorene-co-alt-4,7-(2,1,3-benzothiadiazole) dibromide]), and neutral PNA (peptide nuc...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.307

    authors: Sun C,Gaylord BS,Hong JW,Liu B,Bazan GC

    更新日期:2007-01-01 00:00:00

  • Fluorescence resonance energy transfer microscopy as demonstrated by measuring the activation of the serine/threonine kinase Akt.

    abstract::This protocol describes procedures for performing fluorescence resonance energy transfer (FRET) microscopy analysis by three different methods: acceptor photobleaching, sensitized emission and spectral imaging. We also discuss anisotropy and fluorescence lifetime imaging microscopy-based FRET techniques. By using the ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2012.147

    authors: Broussard JA,Rappaz B,Webb DJ,Brown CM

    更新日期:2013-02-01 00:00:00

  • The touchscreen operant platform for assessing executive function in rats and mice.

    abstract::This protocol details a subset of assays developed within the touchscreen platform to measure various aspects of executive function in rodents. Three main procedures are included: extinction, measuring the rate and extent of curtailing a response that was previously, but is no longer, associated with reward; reversal ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2013.123

    authors: Mar AC,Horner AE,Nilsson SR,Alsiö J,Kent BA,Kim CH,Holmes A,Saksida LM,Bussey TJ

    更新日期:2013-10-01 00:00:00

  • A protocol for isolation and culture of mesenchymal stem cells from mouse compact bone.

    abstract::Unlike humans, mouse bone marrow-derived mesenchymal stem cells (MSCs) cannot be easily harvested by adherence to plastic owing to the contamination of cultures by hematopoietic cells. The design of the protocol described here is based on the phenomenon that compact bones abound in MSCs and hematopoietic cells exist i...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2009.238

    authors: Zhu H,Guo ZK,Jiang XX,Li H,Wang XY,Yao HY,Zhang Y,Mao N

    更新日期:2010-03-01 00:00:00

  • Feeder-free culture of human embryonic stem cells in conditioned medium for efficient genetic modification.

    abstract::Realizing the potential of human embryonic stem cells (hESCs) in research and commercial applications requires generic protocols for culture, expansion and genetic modification that function between multiple lines. Here we describe a feeder-free hESC culture protocol that was tested in 13 independent hESC lines derive...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2008.140

    authors: Braam SR,Denning C,Matsa E,Young LE,Passier R,Mummery CL

    更新日期:2008-01-01 00:00:00

  • Primary culture of bovine chromaffin cells.

    abstract::This protocol describes the primary culture of individual chromaffin cells derived by enzymatic digestion from the adrenal medulla of the bovine adrenal gland. Since the late 1970s, such cells have provided a useful model system to study neurotransmitter biosynthesis, storage and release in the catecholaminergic syste...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.136

    authors: O'Connor DT,Mahata SK,Mahata M,Jiang Q,Hook VY,Taupenot L

    更新日期:2007-01-01 00:00:00

  • Measuring bone blood supply in mice using fluorescent microspheres.

    abstract::Fluorescent microspheres are commonly used to assess bone blood supply in large animals, but the technique is not widely used in smaller mammals, as traditional methods such as reference blood sampling, ventilation and catheterization are not easily applied. This protocol describes a viable alternative for measuring b...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2009.190

    authors: Serrat MA

    更新日期:2009-01-01 00:00:00

  • Derivation and maintenance of human embryonic stem cells from poor-quality in vitro fertilization embryos.

    abstract::Human embryonic stem (hES) cells are self-renewing, pluripotent cells that are valuable research tools and hold promise for use in regenerative medicine. Most hES cell lines are derived from cryopreserved human embryos that were created during in vitro fertilization (IVF) and are in excess of clinical need. Embryos th...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2008.60

    authors: Lerou PH,Yabuuchi A,Huo H,Miller JD,Boyer LF,Schlaeger TM,Daley GQ

    更新日期:2008-01-01 00:00:00

  • Genetically modified mouse models to help fight COVID-19.

    abstract::The research community is in a race to understand the molecular mechanisms of severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) infection, to repurpose currently available antiviral drugs and to develop new therapies and vaccines against coronavirus disease 2019 (COVID-19). One major challenge in achieving ...

    journal_title:Nature protocols

    pub_type: 杂志文章,评审

    doi:10.1038/s41596-020-00403-2

    authors: Gurumurthy CB,Quadros RM,Richardson GP,Poluektova LY,Mansour SL,Ohtsuka M

    更新日期:2020-12-01 00:00:00

  • A protocol for phenotypic detection and characterization of vascular cells of different origins in a lung neovascularization model in rodents.

    abstract::The goal of many current studies of neovascularization is to define the phenotype of vascular cell populations of different origins and to determine how such cells promote assembly of vascular channel. Here, we describe a protocol to immunophenotype vascular cells by high-resolution imaging and by fluorescence-activat...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.537

    authors: Jones RC,Capen DE,Cohen KS,Munn LL,Jain RK,Duda DG

    更新日期:2008-01-01 00:00:00

  • CRISPR-Cas9-mediated genome editing in apple and grapevine.

    abstract::The CRISPR-Cas9 genome-editing tool and the availability of whole-genome sequences from plant species have revolutionized our ability to introduce targeted mutations into important crop plants, both to explore genetic changes and to introduce new functionalities. Here, we describe protocols adapting the CRISPR-Cas9 sy...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-018-0067-9

    authors: Osakabe Y,Liang Z,Ren C,Nishitani C,Osakabe K,Wada M,Komori S,Malnoy M,Velasco R,Poli M,Jung MH,Koo OJ,Viola R,Nagamangala Kanchiswamy C

    更新日期:2018-12-01 00:00:00

  • Affinity purification-mass spectrometry and network analysis to understand protein-protein interactions.

    abstract::By determining protein-protein interactions in normal, diseased and infected cells, we can improve our understanding of cellular systems and their reaction to various perturbations. In this protocol, we discuss how to use data obtained in affinity purification-mass spectrometry (AP-MS) experiments to generate meaningf...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2014.164

    authors: Morris JH,Knudsen GM,Verschueren E,Johnson JR,Cimermancic P,Greninger AL,Pico AR

    更新日期:2014-11-01 00:00:00

  • A general purpose method for extracting RNA from Dictyostelium cells.

    abstract::Here we present a protocol for the extraction of RNA from Dictyostelium discoideum. Dictyostelium is a social amoeba that undergoes a basic developmental program, and therefore analysis of RNA levels over a time course is a commonly used technique. This procedure is similar to other guanidine thiocyanate-based methods...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.191

    authors: Pilcher KE,Gaudet P,Fey P,Kowal AS,Chisholm RL

    更新日期:2007-01-01 00:00:00

  • Accessing crystal-crystal interaction forces with oriented nanocrystal atomic force microscopy probes.

    abstract::Biominerals serve as critical structures of living systems and play important roles in biochemical processes. Understanding their crystallization mechanisms is therefore central to many areas of biology, biogeoscience, and biochemistry. Some biominerals, such as bone and dentin, are hierarchical nanocomposite structur...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/s41596-018-0027-4

    authors: Zhang X,He Y,Liu J,Bowden ME,Kovarik L,Mao SX,Wang C,De Yoreo JJ,Rosso KM

    更新日期:2018-09-01 00:00:00

  • Using the rat forced swim test to assess antidepressant-like activity in rodents.

    abstract::The forced swim test (FST) is one of the most commonly used animal models for assessing antidepressant-like behavior. This protocol details using the FST in rats, which takes place over 48 h and is followed by the video analysis of the behavior. The swim test involves the scoring of active (swimming and climbing) or p...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2012.044

    authors: Slattery DA,Cryan JF

    更新日期:2012-05-03 00:00:00

  • Generation and purification of highly specific antibodies for detecting post-translationally modified proteins in vivo.

    abstract::Post-translational modifications alter protein structure, affecting activity, stability, localization and/or binding partners. Antibodies that specifically recognize post-translationally modified proteins have a number of uses including immunocytochemistry and immunoprecipitation of the modified protein to purify prot...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2014.017

    authors: Arur S,Schedl T

    更新日期:2014-02-01 00:00:00

  • Isolation of human monoclonal antibodies from peripheral blood B cells.

    abstract::Isolation of monoclonal antibodies is an important technique for understanding the specificities and characteristics of antibodies that underlie the humoral immune response to a given antigen. Here we describe a technique for isolating monoclonal antibodies from human peripheral blood mononuclear cells. The protocol i...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2013.117

    authors: Huang J,Doria-Rose NA,Longo NS,Laub L,Lin CL,Turk E,Kang BH,Migueles SA,Bailer RT,Mascola JR,Connors M

    更新日期:2013-10-01 00:00:00

  • Characterizing peptides in individual mammalian cells using mass spectrometry.

    abstract::Cell-to-cell chemical signaling plays multiple roles in coordinating the activity of the functional elements of an organism, with these elements ranging from a three-neuron reflex circuit to the entire animal. In recent years, single-cell mass spectrometry (MS) has enabled the discovery of cell-to-cell signaling molec...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2007.277

    authors: Rubakhin SS,Sweedler JV

    更新日期:2007-01-01 00:00:00

  • Count-based differential expression analysis of RNA sequencing data using R and Bioconductor.

    abstract::RNA sequencing (RNA-seq) has been rapidly adopted for the profiling of transcriptomes in many areas of biology, including studies into gene regulation, development and disease. Of particular interest is the discovery of differentially expressed genes across different conditions (e.g., tissues, perturbations) while opt...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2013.099

    authors: Anders S,McCarthy DJ,Chen Y,Okoniewski M,Smyth GK,Huber W,Robinson MD

    更新日期:2013-09-01 00:00:00

  • A cell-free organelle-based in vitro system for studying the peroxisomal protein import machinery.

    abstract::Here we describe a protocol to dissect the peroxisomal matrix protein import pathway using a cell-free in vitro system. The system relies on a postnuclear supernatant (PNS), which is prepared from rat/mouse liver, to act as a source of peroxisomes and cytosolic components. A typical in vitro assay comprises the follow...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2016.147

    authors: Rodrigues TA,Francisco T,Dias AF,Pedrosa AG,Grou CP,Azevedo JE

    更新日期:2016-12-01 00:00:00

  • A high-yield double-purification proteomics strategy for the identification of SUMO sites.

    abstract::The small ubiquitin-like modifier (SUMO) is a protein modifier that is post-translationally coupled to thousands of lysines in more than a thousand proteins. An understanding of which lysines are modified by SUMO is critical in unraveling its function as a master regulator of all nuclear processes, as well as its invo...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2016.082

    authors: Hendriks IA,Vertegaal AC

    更新日期:2016-09-01 00:00:00

  • High-throughput identification of proteins with AMPylation using self-assembled human protein (NAPPA) microarrays.

    abstract::AMPylation (adenylylation) has been recognized as an important post-translational modification that is used by pathogens to regulate host cellular proteins and their associated signaling pathways. AMPylation has potential functions in various cellular processes, and it is widely conserved across both prokaryotes and e...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2015.044

    authors: Yu X,LaBaer J

    更新日期:2015-05-01 00:00:00

  • Radiolabeling of HYNIC-annexin V with technetium-99m for in vivo imaging of apoptosis.

    abstract::Apoptosis is a critical factor in AIDS and other viral illnesses, cerebral and myocardial ischemia, autoimmune and neurodegenerative states, organ and bone marrow transplant rejection, and tumor response to chemotherapy and radiation. Improved methods to identify sites of apoptosis are increasing our understanding of ...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2006.17

    authors: Blankenberg FG,Vanderheyden JL,Strauss HW,Tait JF

    更新日期:2006-01-01 00:00:00

  • Footprinting protein-DNA complexes using the hydroxyl radical.

    abstract::Hydroxyl radical footprinting has been widely used for studying the structure of DNA and DNA-protein complexes. The high reactivity and lack of base specificity of the hydroxyl radical makes it an excellent probe for high-resolution footprinting of DNA-protein complexes; this technique can provide structural detail th...

    journal_title:Nature protocols

    pub_type: 杂志文章

    doi:10.1038/nprot.2008.72

    authors: Jain SS,Tullius TD

    更新日期:2008-01-01 00:00:00