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  1. Article ; Online: Isolation of Human CD49f+ Astrocytes and In Vitro iPSC-Based Neurotoxicity Assays

    Lilianne Barbar / Tomasz Rusielewicz / Matthew Zimmer / Kriti Kalpana / Valentina Fossati

    STAR Protocols, Vol 1, Iss 3, Pp 100172- (2020)

    2020  

    Abstract: Summary: Given the critical roles of astrocytes in neuroinflammation and neurological diseases, models for studying human astrocyte biology are in increasing demand. Here, we present a protocol to isolate human astrocytes from induced pluripotent stem ... ...

    Abstract Summary: Given the critical roles of astrocytes in neuroinflammation and neurological diseases, models for studying human astrocyte biology are in increasing demand. Here, we present a protocol to isolate human astrocytes from induced pluripotent stem cell (iPSC)-based cultures, neural organoids, and primary tissue, using the surface marker CD49f. Moreover, we provide protocols for in vitro co-cultures of human iPSC-derived neurons and astrocytes, as well as for neurotoxicity assays that expose neurons to conditioned media from reactive astrocytes.For complete details on the use and execution of this protocol, please refer to Barbar et al. (2020).
    Keywords Cell Biology ; Neuroscience ; Stem Cells ; Science (General) ; Q1-390
    Language English
    Publishing date 2020-12-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  2. Article ; Online: Epigenetic Modulation of Human Induced Pluripotent Stem Cell Differentiation to Oligodendrocytes

    Panagiotis Douvaras / Tomasz Rusielewicz / Kwi Hye Kim / Jeffery D. Haines / Patrizia Casaccia / Valentina Fossati

    International Journal of Molecular Sciences, Vol 17, Iss 4, p

    2016  Volume 614

    Abstract: Pluripotent stem cells provide an invaluable tool for generating human, disease-relevant cells. Multiple sclerosis is an inflammatory demyelinating disease of the central nervous system, characterized by myelin damage. Oligodendrocytes are the ... ...

    Abstract Pluripotent stem cells provide an invaluable tool for generating human, disease-relevant cells. Multiple sclerosis is an inflammatory demyelinating disease of the central nervous system, characterized by myelin damage. Oligodendrocytes are the myelinating cells of the central nervous system (CNS); they differentiate from progenitor cells, and their membranes ensheath axons, providing trophic support and allowing fast conduction velocity. The current understanding of oligodendrocyte biology was founded by rodent studies, where the establishment of repressive epigenetic marks on histone proteins, followed by activation of myelin genes, leads to lineage progression. To assess whether this epigenetic regulation is conserved across species, we differentiated human embryonic and induced pluripotent stem cells to oligodendrocytes and asked whether similar histone marks and relative enzymatic activities could be detected. The transcriptional levels of enzymes responsible for methylation and acetylation of histone marks were analyzed during oligodendrocyte differentiation, and the post-translational modifications on histones were detected using immunofluorescence. These studies showed that also in human cells, differentiation along the oligodendrocyte lineage is characterized by the acquisition of multiple repressive histone marks, including deacetylation of lysine residues on histone H3 and trimethylation of residues K9 and K27. These data suggest that the epigenetic modulation of oligodendrocyte identity is highly conserved across species.
    Keywords human induced pluripotent stem cells ; oligodendrocyte differentiation ; histone modifications ; Biology (General) ; QH301-705.5 ; Chemistry ; QD1-999
    Subject code 572
    Language English
    Publishing date 2016-04-01T00:00:00Z
    Publisher MDPI AG
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: Directed Differentiation of Human Pluripotent Stem Cells to Microglia

    Panagiotis Douvaras / Bruce Sun / Minghui Wang / Ilya Kruglikov / Gregory Lallos / Matthew Zimmer / Cecile Terrenoire / Bin Zhang / Sam Gandy / Eric Schadt / Donald O. Freytes / Scott Noggle / Valentina Fossati

    Stem Cell Reports, Vol 8, Iss 6, Pp 1516-

    2017  Volume 1524

    Abstract: Microglia, the immune cells of the brain, are crucial to proper development and maintenance of the CNS, and their involvement in numerous neurological disorders is increasingly being recognized. To improve our understanding of human microglial biology, ... ...

    Abstract Microglia, the immune cells of the brain, are crucial to proper development and maintenance of the CNS, and their involvement in numerous neurological disorders is increasingly being recognized. To improve our understanding of human microglial biology, we devised a chemically defined protocol to generate human microglia from pluripotent stem cells. Myeloid progenitors expressing CD14/CX3CR1 were generated within 30 days of differentiation from both embryonic and induced pluripotent stem cells (iPSCs). Further differentiation of the progenitors resulted in ramified microglia with highly motile processes, expressing typical microglial markers. Analyses of gene expression and cytokine release showed close similarities between iPSC-derived (iPSC-MG) and human primary microglia as well as clear distinctions from macrophages. iPSC-MG were able to phagocytose and responded to ADP by producing intracellular Ca2+ transients, whereas macrophages lacked such response. The differentiation protocol was highly reproducible across several pluripotent stem cell lines.
    Keywords human pluripotent stem cells ; microglial differentiation ; human microglia ; Medicine (General) ; R5-920 ; Biology (General) ; QH301-705.5
    Language English
    Publishing date 2017-06-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  4. Article ; Online: Efficient Generation of Myelinating Oligodendrocytes from Primary Progressive Multiple Sclerosis Patients by Induced Pluripotent Stem Cells

    Panagiotis Douvaras / Jing Wang / Matthew Zimmer / Stephanie Hanchuk / Melanie A. O’Bara / Saud Sadiq / Fraser J. Sim / James Goldman / Valentina Fossati

    Stem Cell Reports, Vol 3, Iss 2, Pp 250-

    2014  Volume 259

    Abstract: Multiple sclerosis (MS) is a chronic demyelinating disease of unknown etiology that affects the CNS. While current therapies are primarily directed against the immune system, the new challenge is to address progressive MS with remyelinating and ... ...

    Abstract Multiple sclerosis (MS) is a chronic demyelinating disease of unknown etiology that affects the CNS. While current therapies are primarily directed against the immune system, the new challenge is to address progressive MS with remyelinating and neuroprotective strategies. Here, we develop a highly reproducible protocol to efficiently derive oligodendrocyte progenitor cells (OPCs) and mature oligodendrocytes from induced pluripotent stem cells (iPSCs). Key elements of our protocol include adherent cultures, dual SMAD inhibition, and addition of retinoids from the beginning of differentiation, which lead to increased yields of OLIG2 progenitors and high numbers of OPCs within 75 days. Furthermore, we show the generation of viral and integration-free iPSCs from primary progressive MS (PPMS) patients and their efficient differentiation to oligodendrocytes. PPMS OPCs are functional, as demonstrated by in vivo myelination in the shiverer mouse. These results provide encouraging advances toward the development of autologous cell therapies using iPSCs.
    Keywords Medicine (General) ; R5-920 ; Biology (General) ; QH301-705.5
    Language English
    Publishing date 2014-08-01T00:00:00Z
    Publisher Elsevier
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  5. Article ; Online: The epichaperome is a mediator of toxic hippocampal stress and leads to protein connectivity-based dysfunction

    Maria Carmen Inda / Suhasini Joshi / Tai Wang / Alexander Bolaender / Srinivasa Gandu / John Koren III / Alicia Yue Che / Tony Taldone / Pengrong Yan / Weilin Sun / Mohammad Uddin / Palak Panchal / Matthew Riolo / Smit Shah / Afsar Barlas / Ke Xu / Lon Yin L. Chan / Alexandra Gruzinova / Sarah Kishinevsky /
    Lorenz Studer / Valentina Fossati / Scott A. Noggle / Julie R. White / Elisa de Stanchina / Sonia Sequeira / Kyle H. Anthoney / John W. Steele / Katia Manova-Todorova / Sujata Patil / Mark P. Dunphy / NagaVaraKishore Pillarsetty / Ana C. Pereira / Hediye Erdjument-Bromage / Thomas A. Neubert / Anna Rodina / Stephen D. Ginsberg / Natalia De Marco Garcia / Wenjie Luo / Gabriela Chiosis

    Nature Communications, Vol 11, Iss 1, Pp 1-

    2020  Volume 19

    Abstract: The biology of Alzheimer’s disease (AD) remains unknown. We propose AD is a protein connectivity-based dysfunction disorder whereby a switch of the chaperome into epichaperomes rewires proteome-wide connectivity, leading to brain circuitry malfunction ... ...

    Abstract The biology of Alzheimer’s disease (AD) remains unknown. We propose AD is a protein connectivity-based dysfunction disorder whereby a switch of the chaperome into epichaperomes rewires proteome-wide connectivity, leading to brain circuitry malfunction that can be corrected by novel therapeutics.
    Keywords Science ; Q
    Language English
    Publishing date 2020-01-01T00:00:00Z
    Publisher Nature Publishing Group
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  6. Article ; Online: The epichaperome is a mediator of toxic hippocampal stress and leads to protein connectivity-based dysfunction

    Maria Carmen Inda / Suhasini Joshi / Tai Wang / Alexander Bolaender / Srinivasa Gandu / John Koren III / Alicia Yue Che / Tony Taldone / Pengrong Yan / Weilin Sun / Mohammad Uddin / Palak Panchal / Matthew Riolo / Smit Shah / Afsar Barlas / Ke Xu / Lon Yin L. Chan / Alexandra Gruzinova / Sarah Kishinevsky /
    Lorenz Studer / Valentina Fossati / Scott A. Noggle / Julie R. White / Elisa de Stanchina / Sonia Sequeira / Kyle H. Anthoney / John W. Steele / Katia Manova-Todorova / Sujata Patil / Mark P. Dunphy / NagaVaraKishore Pillarsetty / Ana C. Pereira / Hediye Erdjument-Bromage / Thomas A. Neubert / Anna Rodina / Stephen D. Ginsberg / Natalia De Marco Garcia / Wenjie Luo / Gabriela Chiosis

    Nature Communications, Vol 11, Iss 1, Pp 1-

    2020  Volume 19

    Abstract: The biology of Alzheimer’s disease (AD) remains unknown. We propose AD is a protein connectivity-based dysfunction disorder whereby a switch of the chaperome into epichaperomes rewires proteome-wide connectivity, leading to brain circuitry malfunction ... ...

    Abstract The biology of Alzheimer’s disease (AD) remains unknown. We propose AD is a protein connectivity-based dysfunction disorder whereby a switch of the chaperome into epichaperomes rewires proteome-wide connectivity, leading to brain circuitry malfunction that can be corrected by novel therapeutics.
    Keywords Science ; Q
    Language English
    Publishing date 2020-01-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

    Full text online

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    Inter-library loan at ZB MED

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