Abstract:
:The fabrication of 3D tissues retaining the original functions of tissues/organs in vitro is crucial for optimal tissue engineering and regenerative medicine. The fabrication of 3D tissues also contributes to the establishment of in vitro tissue/organ models for drug screening. Our laboratory has developed a fabrication system for functional 3D tissues by stacking cell sheets of confluent cultured cells detached from a temperature-responsive culture dish. Here we describe the protocols for the fabrication of 3D tissues by cell sheet engineering. Three-dimensional cardiac tissues fabricated by stacking cardiac cell sheets pulsate spontaneously, synchronously and macroscopically. Via this protocol, it is also possible to fabricate other tissues, such as 3D tissue including capillary-like prevascular networks, from endothelial cells sandwiched between layered cell sheets. Cell sheet stacking technology promises to provide in vitro tissue/organ models and more effective therapies for curing tissue/organ failures.
journal_name
Nat Protocjournal_title
Nature protocolsauthors
Haraguchi Y,Shimizu T,Sasagawa T,Sekine H,Sakaguchi K,Kikuchi T,Sekine W,Sekiya S,Yamato M,Umezu M,Okano Tdoi
10.1038/nprot.2012.027subject
Has Abstractpub_date
2012-04-05 00:00:00pages
850-8issue
5eissn
1754-2189issn
1750-2799pii
nprot.2012.027journal_volume
7pub_type
杂志文章相关文献
Nature Protocols文献大全abstract::The world is on the cusp of a post-antibiotic era, but researchers and medical doctors have found a way forward-by looking back at how infections were treated before the advent of antibiotics, namely using phage therapy. Although bacteriophages (phages) continue to lack drug approval in Western medicine, an increasing...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-020-0346-0
更新日期:2020-09-01 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::The dynamics of transcription can be studied genome wide by high-throughput sequencing of nascent and newly synthesized RNA. 4-thiouridine (4SU) labeling in vivo enables the specific capture of such new transcripts, with 4SU residues being tagged by biotin linkers and captured using streptavidin beads before library p...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-019-0262-3
更新日期:2020-02-01 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::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
更新日期:2018-09-01 00:00:00
abstract::The ability to isolate and analyze rare circulating tumor cells (CTCs) has the potential to further our understanding of cancer metastasis and enhance the care of cancer patients. In this protocol, we describe the procedure for isolating rare CTCs from blood samples by using tumor antigen-independent microfluidic CTC-...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.044
更新日期:2014-03-01 00:00:00
abstract::The analysis of large microbiome data sets holds great promise for the delineation of the biological and metabolic functioning of living organisms and their role in the environment. In the midst of this genomic puzzle, viruses, especially those that infect microbial communities, represent a major reservoir of genetic ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2017.063
更新日期:2017-08-01 00:00:00
abstract::Cyclodepsipeptides are cyclic peptides in which at least one amide link on the backbone is replaced with an ester link. These natural products present a high structural diversity that corresponds to a broad range of biological activities. Therefore, they are very promising pharmaceutical candidates. Most of the cyclod...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2016.116
更新日期:2016-10-01 00:00:00
abstract::Mass spectrometry imaging (MSI) enables label-free spatial mapping of hundreds of biomolecules in tissue sections. This capability provides valuable information on tissue heterogeneity that is difficult to obtain using population-averaged assays. Despite substantial developments in both instrumentation and methodology...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-019-0237-4
更新日期:2019-12-01 00:00:00
abstract::Collagen-mimetic peptides and lipopeptides are widely used as substrates for matrix degrading enzymes, as new biomaterials for tissue engineering, as drug delivery systems and so on. However, the preparation and subsequent purification of these peptides and their fatty-acid conjugates are really challenging. Herein, w...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.195
更新日期:2010-01-01 00:00:00
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
更新日期:2015-09-01 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::RNase H-dependent PCR-enabled T-cell receptor sequencing (rhTCRseq) can be used to determine paired alpha/beta T-cell receptor (TCR) clonotypes in single cells or perform alpha and beta TCR repertoire analysis in bulk RNA samples. With the enhanced specificity of RNase H-dependent PCR (rhPCR), it achieves TCR-specific...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-019-0195-x
更新日期:2019-08-01 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::An important area of proteomics involves the need for quantification, whether relative or absolute. Many methods now exist for relative quantification, but to support biomarker proteomics and systems biology, absolute quantification rather than relative quantification is required. Absolute quantification usually invol...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.129
更新日期:2006-01-01 00:00:00
abstract::Neurotransmitter release is triggered by membrane depolarization, Ca(2+) influx and Ca(2+) sensing by the release machinery, causing synaptic vesicle (SV) fusion with the plasma membrane. Interlinked is a complex membrane cycle in which vesicles are tethered to the release site, primed, fused and recycled. As many of ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2012.074
更新日期:2012-06-21 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::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::Protein ADP-ribosylation is a structurally heterogeneous post-translational modification (PTM) that influences the physicochemical and biological properties of the modified protein. ADP-ribosylation of chromatin changes its structural properties, thereby regulating important nuclear functions. A lack of suitable antib...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2017.072
更新日期:2017-09-01 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
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::A mechanistic understanding of HIV-1 latency depends on a model system that recapitulates the in vivo condition of latently infected, resting CD4(+) T lymphocytes. Latency seems to be established after activated CD4(+) T cells, the principal targets of HIV-1 infection, become productively infected and survive long eno...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2014.188
更新日期:2014-12-01 00:00:00
abstract::The combination of ChIP-seq and transcriptome analysis is a compelling approach to unravel the regulation of gene expression. Several recently published methods combine transcription factor (TF) binding and gene expression for target prediction, but few of them provide an efficient software package for the community. ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2013.150
更新日期:2013-12-01 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::Here, we describe a robust protocol for the reverse transfection of cells on small interfering (siRNA) arrays, which, in combination with multi-channel immunofluorescence or time-lapse microscopy, is suitable for genome-wide RNA interference (RNAi) screens in intact human cells. The automatic production of 48 'transfe...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.483
更新日期:2007-01-01 00:00:00
abstract::Freestanding plasmonic nanoparticle (NP) superlattice sheets are novel 2D nanomaterials with tailorable properties that enable their use for broad applications in sensing, anticounterfeit measures, ionic gating, nanophotonics and flat lenses. We recently developed a robust, yet general, two-step drying-mediated approa...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/s41596-019-0200-4
更新日期:2019-09-01 00:00:00
abstract::Here, we describe a simple micromolding method to construct three-dimensional arrays of organotypic epithelial tissue structures that approximate in vivo histology. An elastomeric stamp containing an array of posts of defined geometry and spacing is used to mold microscale cavities into the surface of type I collagen ...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.35
更新日期:2008-01-01 00:00:00
abstract::The effect of genetic mutation on phenotype is of significant interest in genetics. The type of genetic mutation that causes a single amino acid substitution (AAS) in a protein sequence is called a non-synonymous single nucleotide polymorphism (nsSNP). An nsSNP could potentially affect the function of the protein, sub...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2009.86
更新日期:2009-01-01 00:00:00
abstract::In this report we describe the development of a standardized three-dimensional (3D) system of the human oral mucosa based on an immortalized human oral keratinocyte cell line (OKF6/TERT-2). The procedure takes approximately 2-3 weeks to complete and includes three main stages: preparation of collagen-embedded fibrobla...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2006.323
更新日期:2006-01-01 00:00:00
abstract::Unlike transplantation of other solid organs, vascularized mouse lung transplantation has only recently been developed. In this protocol, we describe a detailed method for performing a vascularized and aerated mouse orthotopic lung transplant, which to date represents the most physiological mouse model of lung transpl...
journal_title:Nature protocols
pub_type: 杂志文章
doi:10.1038/nprot.2008.218
更新日期:2009-01-01 00:00:00