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  • 1
    Publication Date: 2013-06-04
    Description: The generation of induced pluripotent stem (iPS) cells presents a challenge to normal developmental processes. The low efficiency and heterogeneity of most methods have hindered understanding of the precise molecular mechanisms promoting, and roadblocks preventing, efficient reprogramming. Although several intermediate populations have been described, it has proved difficult to characterize the rare, asynchronous transition from these intermediate stages to iPS cells. The rapid expansion of minor reprogrammed cells in the heterogeneous population can also obscure investigation of relevant transition processes. Understanding the biological mechanisms essential for successful iPS cell generation requires both accurate capture of cells undergoing the reprogramming process and identification of the associated global gene expression changes. Here we demonstrate that in mouse embryonic fibroblasts, reprogramming follows an orderly sequence of stage transitions, marked by changes in the cell-surface markers CD44 and ICAM1, and a Nanog-enhanced green fluorescent protein (Nanog-eGFP) reporter. RNA-sequencing analysis of these populations demonstrates two waves of pluripotency gene upregulation, and unexpectedly, transient upregulation of several epidermis-related genes, demonstrating that reprogramming is not simply the reversal of the normal developmental processes. This novel high-resolution analysis enables the construction of a detailed reprogramming route map, and the improved understanding of the reprogramming process will lead to new reprogramming strategies.〈br /〉〈br /〉〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3743022/" target="_blank"〉〈img src="https://static.pubmed.gov/portal/portal3rc.fcgi/4089621/img/3977009" border="0"〉〈/a〉   〈a href="https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3743022/" target="_blank"〉This paper as free author manuscript - peer-reviewed and accepted for publication〈/a〉〈br /〉〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉O'Malley, James -- Skylaki, Stavroula -- Iwabuchi, Kumiko A -- Chantzoura, Eleni -- Ruetz, Tyson -- Johnsson, Anna -- Tomlinson, Simon R -- Linnarsson, Sten -- Kaji, Keisuke -- 261075/European Research Council/International -- Medical Research Council/United Kingdom -- England -- Nature. 2013 Jul 4;499(7456):88-91. doi: 10.1038/nature12243. Epub 2013 Jun 2.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉MRC Centre for Regenerative Medicine, University of Edinburgh, Edinburgh BioQuarter, 5 Little France Drive, Edinburgh EH16 4UU, UK.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/23728301" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; Antigens, CD44/genetics/*metabolism ; Biomarkers/analysis/metabolism ; Cellular Reprogramming/genetics/*physiology ; Epidermis/metabolism ; Fibroblasts ; Flow Cytometry ; Gene Expression Profiling ; Genes, Reporter ; Induced Pluripotent Stem Cells/*cytology/*metabolism ; Intercellular Adhesion Molecule-1/genetics/*metabolism ; Mice ; Sequence Analysis, RNA ; Single-Cell Analysis ; Up-Regulation/genetics
    Print ISSN: 0028-0836
    Electronic ISSN: 1476-4687
    Topics: Biology , Chemistry and Pharmacology , Medicine , Natural Sciences in General , Physics
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  • 2
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    American Association for the Advancement of Science (AAAS)
    Publication Date: 2015-10-03
    Description: 〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Linnarsson, Sten -- New York, N.Y. -- Science. 2015 Oct 2;350(6256):37. doi: 10.1126/science.aad2792.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-17177 Stockholm, Sweden. sten.linnarsson@ki.se.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/26430106" target="_blank"〉PubMed〈/a〉
    Keywords: Cerebral Cortex/*cytology/*growth & development ; Female ; Humans ; Male ; *Mutation ; Neurons/*cytology/*physiology ; *Polymorphism, Single Nucleotide ; *Transcription, Genetic
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 3
    Publication Date: 2015-02-24
    Description: The mammalian cerebral cortex supports cognitive functions such as sensorimotor integration, memory, and social behaviors. Normal brain function relies on a diverse set of differentiated cell types, including neurons, glia, and vasculature. Here, we have used large-scale single-cell RNA sequencing (RNA-seq) to classify cells in the mouse somatosensory cortex and hippocampal CA1 region. We found 47 molecularly distinct subclasses, comprising all known major cell types in the cortex. We identified numerous marker genes, which allowed alignment with known cell types, morphology, and location. We found a layer I interneuron expressing Pax6 and a distinct postmitotic oligodendrocyte subclass marked by Itpr2. Across the diversity of cortical cell types, transcription factors formed a complex, layered regulatory code, suggesting a mechanism for the maintenance of adult cell type identity.〈br /〉〈span class="detail_caption"〉Notes: 〈/span〉Zeisel, Amit -- Munoz-Manchado, Ana B -- Codeluppi, Simone -- Lonnerberg, Peter -- La Manno, Gioele -- Jureus, Anna -- Marques, Sueli -- Munguba, Hermany -- He, Liqun -- Betsholtz, Christer -- Rolny, Charlotte -- Castelo-Branco, Goncalo -- Hjerling-Leffler, Jens -- Linnarsson, Sten -- New York, N.Y. -- Science. 2015 Mar 6;347(6226):1138-42. doi: 10.1126/science.aaa1934. Epub 2015 Feb 19.〈br /〉〈span class="detail_caption"〉Author address: 〈/span〉Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm, Sweden. ; Department of Immunology, Genetics and Pathology, Rudbeck Laboratory, Uppsala University, Dag Hammarskjolds vag 20, S-751 85 Uppsala, Sweden. ; Department of Immunology, Genetics and Pathology, Rudbeck Laboratory, Uppsala University, Dag Hammarskjolds vag 20, S-751 85 Uppsala, Sweden. Division of Vascular Biology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm, Sweden. ; Department of Oncology-Pathology, Karolinska Institutet, S-171 76 Stockholm, Sweden. ; Division of Molecular Neurobiology, Department of Medical Biochemistry and Biophysics, Karolinska Institutet, S-171 77 Stockholm, Sweden. sten.linnarsson@ki.se jens.hjerling-leffler@ki.se.〈br /〉〈span class="detail_caption"〉Record origin:〈/span〉 〈a href="http://www.ncbi.nlm.nih.gov/pubmed/25700174" target="_blank"〉PubMed〈/a〉
    Keywords: Animals ; CA1 Region, Hippocampal/*cytology/metabolism ; Eye Proteins/genetics ; Gene Expression ; Genetic Markers ; Homeodomain Proteins/genetics ; Inositol 1,4,5-Trisphosphate Receptors/genetics ; Interneurons/*classification/cytology/metabolism ; Mice ; Oligodendroglia/*classification/cytology/metabolism ; Paired Box Transcription Factors/genetics ; Phylogeny ; Repressor Proteins/genetics ; Sequence Analysis, RNA/*methods ; Single-Cell Analysis/*methods ; Somatosensory Cortex/*cytology/metabolism ; Transcription Factors/classification/genetics ; Transcriptome
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 4
    Publication Date: 2001-11-27
    Print ISSN: 0027-8424
    Electronic ISSN: 1091-6490
    Topics: Biology , Medicine , Natural Sciences in General
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  • 5
    Publication Date: 2010-08-24
    Print ISSN: 0027-8424
    Electronic ISSN: 1091-6490
    Topics: Biology , Medicine , Natural Sciences in General
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  • 6
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    American Association for the Advancement of Science (AAAS)
    In: Science
    Publication Date: 2017-10-06
    Description: The stereotyped spatial architecture of the brain is both beautiful and fundamentally related to its function, extending from gross morphology to individual neuron types, where soma position, dendritic architecture, and axonal projections determine their roles in functional circuitry. Our understanding of the cell types that make up the brain is rapidly accelerating, driven in particular by recent advances in single-cell transcriptomics. However, understanding brain function, development, and disease will require linking molecular cell types to morphological, physiological, and behavioral correlates. Emerging spatially resolved transcriptomic methods promise to fill this gap by localizing molecularly defined cell types in tissues, with simultaneous detection of morphology, activity, or connectivity. Here, we review the requirements for spatial transcriptomic methods toward these goals, consider the challenges ahead, and describe promising applications.
    Keywords: Genetics
    Print ISSN: 0036-8075
    Electronic ISSN: 1095-9203
    Topics: Biology , Chemistry and Pharmacology , Geosciences , Computer Science , Medicine , Natural Sciences in General , Physics
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  • 7
    Publication Date: 2013-10-30
    Description: Motivation: Recent transcriptome studies have revealed that total transcript numbers vary by cell type and condition; therefore, the statistical assumptions for single-cell transcriptome studies must be revisited. SAMstrt is an extension code for SAMseq, which is a statistical method for differential expression, to enable spike-in normalization and statistical testing based on the estimated absolute number of transcripts per cell for single-cell RNA-seq methods. Availability and Implementation: SAMstrt is implemented on R and available in github ( https://github.com/shka/R-SAMstrt ). Contact: shintaro.katayama@ki.se Supplementary Information: Supplementary data are available at Bioinformatics online.
    Print ISSN: 1367-4803
    Electronic ISSN: 1460-2059
    Topics: Biology , Computer Science , Medicine
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