Abstract：
:Establishing a small animal model that accurately recapitulates hepatotropic pathogens, including hepatitis C virus (HCV) infection and immunopathogenesis, is essential for the study of hepatitis virus-induced liver disease and for therapeutics development. This protocol describes our recently developed humanized mouse model for studying HCV and other hepatotropic infections, human immune response and hepatitis and liver fibrosis. The first 5-h stage is the isolation of human liver progenitor and hematopoietic stem cells from fetal liver. Next, AFC8 immunodeficient mice are transplanted with the isolated progenitor/stem cells. This generally takes 2 h. The transplanted mice are then treated for a month with the mouse liver apoptosis-inducing AFC8 dimerizer and left for an additional 2-month period to permit human liver and immune cell growth as well as system reconstitution and development before inoculation with HCV clinical isolates. HCV infection, human immune response and liver disease are observed with high incidence from approximately 2 months after inoculation.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Bility MT,Zhang L,Washburn ML,Curtis TA,Kovalev GI,Su Ldoi
10.1038/nprot.2012.083subject
Has Abstractpub_date
2012-09-01 00:00:00pages
1608-17issue
9eissn
1754-2189issn
1750-2799pii
nprot.2012.083journal_volume
7pub_type
杂志文章相关文献
Nature Protocols文献大全abstract：:The Strep-tag II is an eight-residue minimal peptide sequence (Trp-Ser-His-Pro-Gln-Phe-Glu-Lys) that exhibits intrinsic affinity toward streptavidin and can be fused to recombinant proteins in various fashions. We describe a protocol that enables quick and mild purification of corresponding Strep-tag II fusion protein...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.209
更新日期：2007-01-01 00:00:00
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
更新日期：2008-01-01 00:00:00
abstract：:Pseudo-afferent cervical lymph-duct cannulation in a sheep model allows large amounts of lymph cells to be collected under physiological conditions, carrying immune signaling information from the head tissues, including oro-nasal mucosae. Importantly, large quantities of dendritic cells (DCs) of several subtypes are o...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.147
更新日期：2006-01-01 00:00:00
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
更新日期：2008-01-01 00:00:00
abstract：:Cap-analysis gene expression (CAGE) provides accurate high-throughput measurement of RNA expression. CAGE allows mapping of all the initiation sites of both capped coding and noncoding RNAs. In addition, transcriptional start sites within promoters are characterized at single-nucleotide resolution. The latter allows t...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2012.005
更新日期：2012-02-23 00:00:00
abstract：:Characterization of the cellular participants in tissue immune responses is crucial to understanding infection, cancer, autoimmunity, allergy, graft rejection and other immunological processes. Previous reports indicate that leukocytes in lung vasculature fail to be completely removed by perfusion. Several studies sug...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2014.005
更新日期：2014-01-01 00:00:00
abstract：:RNA interference (RNAi) is an efficient method for silencing genes in cultured cells. Here we describe a simple RNAi approach for silencing genes in a cell type-specific and tissue-specific way in vivo. The approach, which mimics the means by which naturally occurring 'microRNA's are generated, uses a tissue-specific ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.260
更新日期：2006-01-01 00:00:00
abstract：:This protocol outlines the carboxyfluorescein diacetate succinimidyl ester (CFSE) method for following the proliferation of human lymphocytes in vitro and mouse lymphocytes both in vitro and in vivo. The method relies on the ability of CFSE to covalently label long-lived intracellular molecules with the highly fluores...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.296
更新日期：2007-01-01 00:00:00
abstract：:With the advent of massively parallel sequencing, considerable work has gone into adapting chromosome conformation capture (3C) techniques to study chromosomal architecture at a genome-wide scale. We recently demonstrated that the inactive murine X chromosome adopts a bipartite structure using a novel 3C protocol, ter...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.126
更新日期：2016-11-01 00:00:00
abstract：:Spectroscopic techniques such as Fourier-transform infrared (FTIR) spectroscopy are used to study interactions of light with biological materials. This interaction forms the basis of many analytical assays used in disease screening/diagnosis, microbiological studies, and forensic/environmental investigations. Advantag...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/s41596-019-0150-x
更新日期：2019-05-01 00:00:00
abstract：:One of the challenges for modern neuroscience is to understand the rules of concerted neuronal function in vivo. This question can be addressed using noninvasive high-resolution imaging techniques like two-photon microscopy. This protocol describes a versatile approach for in vivo two-photon calcium imaging of neural ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.58
更新日期：2006-01-01 00:00:00
abstract：:MicroRNAs (miRNAs) are short, about 21 nucleotides in length, noncoding, regulatory RNA molecules representing a new layer in post-transcriptional regulation of gene expression. Intensive miRNA research has necessitated the development of effective miRNA detection methods such as northern analyses, quantitative real-t...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.528
更新日期：2008-01-01 00:00:00
abstract：:We present a detailed protocol for the structural analysis of protein-linked glycans. In this approach, appropriate for glycomics studies, N-linked glycans are released using peptide N-glycosidase F and O-linked glycans are released by reductive alkaline beta-elimination. Using strategies based on mass spectrometry (m...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2007.227
更新日期：2007-01-01 00:00:00
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 t...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2009.157
更新日期：2009-01-01 00:00:00
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
更新日期：2016-12-01 00:00:00
abstract：:This protocol describes how to purify and radiolabel collagen for use as a substrate to assay collagenolytic members of the matrix metalloproteinases (MMPs). This assay measures enzymes that specifically cleave native triple helical collagen. After incubation of the MMP enzyme with the collagen substrate at 37 degrees...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2008.244
更新日期：2009-01-01 00:00:00
abstract：:Multiplex-fluorescence in situ hybridization (M-FISH) was initially developed to stain human chromosomes--the 22 autosomes and X and Y sex chromosomes--with uniquely distinctive colors to facilitate karyotyping. The characteristic spectral signatures of all different combinations of fluorochromes are determined by mul...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.160
更新日期：2006-01-01 00:00:00
abstract：:Mouse embryonic stem cells (mESCs) are key tools for genetic engineering, development of stem cell-based therapies and basic research on pluripotency and early lineage commitment. However, successful derivation of germline-competent embryonic stem cell lines has, until recently, been limited to a small number of inbre...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2014.030
更新日期：2014-03-01 00:00:00
abstract：:Single-cell gene expression analysis has contributed to a better understanding of the transcriptional heterogeneity in a variety of model systems, including those used in research in developmental, cancer and stem cell biology. Nowadays, technological advances facilitate the generation of large gene expression data se...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2015.022
更新日期：2015-03-01 00:00:00
abstract：:The rodent malaria parasite Plasmodium chabaudi chabaudi shares many features with human malaria species, including P. falciparum, and is the in vivo model of choice for many aspects of malaria research in the mammalian host, from sequestration of parasitized erythrocytes, to antigenic variation and host immunity and ...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2011.313
更新日期：2011-04-01 00:00:00
abstract：:Generation of precisely patterned neural cells from human pluripotent stem cells (hPSCs) is instrumental in developing disease models and stem cell therapies. Here, we provide a detailed 16-d protocol for obtaining high-purity ventral midbrain (VM) dopamine (DA) progenitors for intracerebral transplantation into anima...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2017.078
更新日期：2017-09-01 00:00:00
abstract：:This protocol details a method for obtaining a considerable number of human mast cells from peripheral blood cells from normal donors without using stem cell mobilization treatment. By using the magnetic cell sorting system, 10(4)-10(5) cells are retrieved in the CD34+ fraction when 100 ml of blood is drawn from a hea...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.344
更新日期：2006-01-01 00:00:00
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
更新日期：2015-05-01 00:00:00
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
更新日期：2012-05-03 00:00:00
abstract：:Aptamers are single-stranded DNA or RNA molecules that can bind target molecules with high affinity and specificity. The conformation of an aptamer usually changes upon binding to its target analyte, and this property has been used in a wide variety of sensing applications, including detection based on fluorescence in...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.38
更新日期：2006-01-01 00:00:00
abstract：:This protocol provides a detailed procedure for the chemical synthesis of proteins through native chemical ligation of peptide hydrazides. The two crucial stages of this protocol are (i) the solid-phase synthesis of peptide hydrazides via Fmoc chemistry and (ii) the native chemical ligation of peptide hydrazides throu...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2013.152
更新日期：2013-12-01 00:00:00
abstract：:The small ubiquitin-like modifiers (SUMOs) are posttranslationally conjugated to eukaryotic cellular proteins with generally unpredictable consequences. SUMO substrates are found in many cellular systems, and functional analysis has revealed that substrate SUMOylation often has an important role in their regulation. H...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2009.128
更新日期：2009-01-01 00:00:00
abstract：:Boronic acids and esters have critical roles in the areas of synthetic organic chemistry, molecular sensors, materials science, drug discovery, and catalysis. Many of the current applications of boronic acids and esters require materials with very low levels of transition metal contamination. Most of the current metho...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2016.184
更新日期：2017-03-01 00:00:00
abstract：:Forebrain γ-aminobutyric acid (GABA) interneurons have crucial roles in high-order brain function via modulating network activities and plasticity, and they are implicated in many psychiatric disorders. Availability of enriched functional human forebrain GABA interneurons, especially those from people affected by GABA...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2013.106
更新日期：2013-09-01 00:00:00
abstract：:The synthesis of an azobenzene amino acid (aa) for use as a photo-inducible conformational switch in polypeptides is described. The compound can be easily incorporated into an aa sequence by solid-phase peptide synthesis using standard 9-fluorenylmethoxycarbonyl methods. A reversible conformational change of the pepti...
journal_title：Nature protocols
pub_type： 杂志文章
doi：10.1038/nprot.2006.488
更新日期：2007-01-01 00:00:00
