Abstract:
:Differentiation of stem cells to hepatocytes has industrial applications, as well as the potential to develop new therapeutic strategies for liver disease. The protocol described here, sequentially using cytokines that are known to have a role in liver embryonic development, efficiently differentiates rat multipotent adult progenitor cells (rMAPCs) to hepatocyte-like cells by directing them through defined embryonic intermediates, namely, primitive streak/mesendoderm/definitive endoderm, hepatoblast and hepatocyte-like phenotype. After 20 days, the final differentiated multipotent adult progenitor cell progeny is a mixture of cells, comprising cells with the characteristics of hepatoblasts and a smaller cell fraction with the morphological and phenotypical features of mature hepatocytes, as well as other mesodermal cells and some persistent undifferentiated rMAPCs. A detailed functional characterization of the stem cell progeny is also described; this should be used to confirm that differentiated cells display the functional characteristics of mature hepatocytes, including albumin secretion, glycogen storage and several detoxifying functions such as urea production, bilirubin conjugation, glutathione S-transferase activity and cytochrome activity.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Roelandt P,Sancho-Bru P,Pauwelyn K,Verfaillie Cdoi
10.1038/nprot.2010.80subject
Has Abstractpub_date
2010-07-01 00:00:00pages
1324-36issue
7eissn
1754-2189issn
1750-2799pii
nprot.2010.80journal_volume
5pub_type
杂志文章相关文献
Nature Protocols文献大全abstract::We describe a step-by-step protocol for measuring the stable products of the nitric oxide (NO) pathway: nitrite, nitrite plus nitrate and nitrate. This described protocol is easy to apply and is about 50 times more sensitive than the commonly used Griess reaction or commercially available assay kits based on the Gries...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.341
更新日期:2006-01-01 00:00:00
abstract::Surface- and matrix-bound signals modulate stem cell fate in vivo and in vitro. This protocol enables the immobilization of a wide range of biomolecules that contain primary amino groups to different types of solid carriers, including glass substrates and standard polystyrene well plates. We describe how thin polymer ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2010.70
更新日期:2010-06-01 00:00:00
abstract::Conventional gene expression studies analyze multiple cells simultaneously or single cells, for which the exact in vivo or in situ position is unknown. Although cellular heterogeneity can be discerned when analyzing single cells, any spatially defined attributes that underpin the heterogeneous nature of the cells cann...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2017.003
更新日期:2017-03-01 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::Herein we describe a protocol that uses hollow-fiber flow field-flow fractionation (FFF) coupled with multiangle light scattering (MALS) for hydrodynamic size-based separation and characterization of complex protein aggregates. The fractionation method, which requires 1.5 h to run, was successfully modified from the a...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.009
更新日期:2015-01-01 00:00:00
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
更新日期:2018-12-01 00:00:00
abstract::The telomeric repeat amplification protocol (TRAP) is a two-step process for analyzing telomerase activity in cell or tissue extracts. Recent modifications of this sensitive assay include elimination of radioactivity by using a fluorescently labeled primer instead of a radiolabeled primer. In addition, the TRAP assay ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.239
更新日期:2006-01-01 00:00:00
abstract::Using siRNAs to genetically manipulate immune cells is important to both basic immunological studies and therapeutic applications. However, siRNA delivery is challenging because primary immune cells are often sensitive to the delivery materials and generate immune responses. We have recently developed an amphiphilic d...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-020-00418-9
更新日期:2021-01-01 00:00:00
abstract::Desorption electrospray ionization (DESI) allows the direct analysis of ordinary objects or pre-processed samples under ambient conditions. Among other applications, DESI is used to identify and record spatial distributions of lipids and drug molecules in biological tissue sections. This technique does not require sam...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.11
更新日期:2008-01-01 00:00:00
abstract::New technologies have recently enabled saturation mutagenesis and functional analysis of nearly all possible variants of regulatory elements or proteins of interest in single experiments. Here we discuss the past, present, and future of such multiplexed (functional) assays for variant effects (MAVEs). MAVEs provide de...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2016.135
更新日期:2016-10-01 00:00:00
abstract::This protocol describes an optimized method for direct in vitro monitoring of homo- and heterotypic axon-axon interactions involved in the developmental assembly of neural circuits. The assay exploits a classical example of heterotypic axonal interactions by modeling the sequential extension of spinal motor and somato...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2011.442
更新日期:2012-01-26 00:00:00
abstract::The tracking of cell fate, shape and migration is an essential component in the study of the development of multicellular organisms. Here we report a protocol that uses the protein Kaede, which is fluorescent green after synthesis but can be photoconverted red by violet or UV light. We have used Kaede along with confo...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.96
更新日期:2006-01-01 00:00:00
abstract::The elucidation of protein-protein interaction networks is a crucial task in the postgenomic era. In this protocol, we describe our approach to discover protein-protein interactions using the surface plasmon resonance technique coupled to mass spectrometry (MS). A peptide or a protein is immobilized on a sensor chip a...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.84
更新日期:2009-01-01 00:00:00
abstract::Single-cell electroporation allows transfection of plasmid DNA or macromolecules into individual living cells using modified patch electrodes and common electrophysiological equipment. This protocol is optimized for rapid in vivo electroporation of Xenopus laevis tadpole brains with DNA, dextrans, morpholinos and comb...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.186
更新日期:2006-01-01 00:00:00
abstract::We describe the implementation and use of an adaptive imaging framework for optimizing spatial resolution and signal strength in a light-sheet microscope. The framework, termed AutoPilot, comprises hardware and software modules for automatically measuring and compensating for mismatches between light-sheet and detecti...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-018-0043-4
更新日期:2018-11-01 00:00:00
abstract::The cytokinesis-block micronucleus cytome assay is a comprehensive system for measuring DNA damage, cytostasis and cytotoxicity. DNA damage events are scored specifically in once-divided binucleated (BN) cells and include (a) micronuclei (MNi), a biomarker of chromosome breakage and/or whole chromosome loss, (b) nucle...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2007.77
更新日期:2007-01-01 00:00:00
abstract::Raman and Fourier transform IR (FTIR) microspectroscopic images of biological material (tissue sections) contain detailed information about their chemical composition. The challenge lies in identifying changes in chemical composition, as well as locating and assigning these changes to different conditions (pathology, ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2015.008
更新日期:2015-02-01 00:00:00
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
更新日期:2009-01-01 00:00:00
abstract::The transmembrane (TM) anchors of cell surface proteins have been one of the 'blind spots' in structural biology because they are generally very hydrophobic, sometimes dynamic, and thus difficult targets for structural characterization. A plethora of examples show these membrane anchors are not merely anchors but can ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-019-0188-9
更新日期:2019-08-01 00:00:00
abstract::Association studies can focus on candidate gene(s), a particular genomic region, or adopt a genome-wide association approach, each of which has implications for marker selection. The strategy for marker selection will affect the statistical power of the study to detect a disease association and is a crucial element of...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.38
更新日期:2009-01-01 00:00:00
abstract::Microfluidic mixing in combination with single-molecule spectroscopy allows the investigation of complex biomolecular processes under non-equilibrium conditions. Here we present a protocol for building, installing and operating microfluidic mixing devices optimized for this purpose. The mixer is fabricated by replica ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.082
更新日期:2013-08-01 00:00:00
abstract::We present a protocol for visualizing and quantifying single mRNA molecules in mammalian (mouse and human) tissues. In the approach described here, sets of about 50 short oligonucleotides, each labeled with a single fluorophore, are hybridized to target mRNAs in tissue sections. Each set binds to a single mRNA molecul...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.109
更新日期:2013-09-01 00:00:00
abstract::Mass spectrometry (MS)-based proteomics has become the preferred tool for the analysis of protein phosphorylation. To be successful at such an endeavor, there is a requirement for an efficient enrichment of phosphopeptides. This is necessary because of the substoichiometric nature of phosphorylation at a given site an...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.010
更新日期:2013-03-01 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::Growing evidence indicates that RNA G-quadruplexes have important roles in various processes such as transcription, translation, regulation of telomere length, and formation of telomeric heterochromatin. Investigation of RNA G-quadruplex structures associated with biological events is therefore essential to understand...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2017.156
更新日期:2018-04-01 00:00:00
abstract::High-throughput ballistic injection nanorheology is a method for the quantitative study of cell mechanics. Cell mechanics are measured by ballistic injection of submicron particles into the cytoplasm of living cells and tracking the spontaneous displacement of the particles at high spatial resolution. The trajectories...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2011.436
更新日期:2012-01-05 00:00:00
abstract::The bimolecular fluorescence complementation (BiFC) assay is a powerful tool for visualizing and identifying protein interactions in living cells. This assay is based on the principle of protein-fragment complementation, using two nonfluorescent fragments derived from fluorescent proteins. When two fragments are broug...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.16
更新日期:2008-01-01 00:00:00
abstract::Top-down proteomics (TDP) by mass spectrometry (MS) is a technique by which intact proteins are analyzed. It has become increasingly popDesalting and concentrating GELFrEEular in translational research because of the value of characterizing distinct proteoforms of intact proteins. Compared to bottom-up proteomics (BUP...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-018-0085-7
更新日期:2019-01-01 00:00:00
abstract::Reactive oxygen species (ROS) are continuously produced in the cell as a consequence of aerobic metabolism, and are controlled by several antioxidant mechanisms. An accurate measurement of ROS is essential to evaluate the redox status of the cell, or the effects of molecules with the pro-oxidant or antioxidant activit...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.189
更新日期:2009-01-01 00:00:00
abstract::During biosynthesis on the ribosome, an elongating nascent polypeptide chain can begin to fold, in a process that is central to all living systems. Detailed structural studies of co-translational protein folding are now beginning to emerge; such studies were previously limited, at least in part, by the inherently dyna...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2016.101
更新日期:2016-08-01 00:00:00