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  1. Article: OCT4 induces long-lived dedifferentiated kidney progenitors poised to redifferentiate in 3D kidney spheroids.

    Omer, Dorit / Zontag, Osnat Cohen / Gnatek, Yehudit / Harari-Steinberg, Orit / Pleniceanu, Oren / Namestnikov, Michael / Cohen, Ayelet-Hashahar / Nissim-Rafinia, Malka / Tam, Gal / Kalisky, Tomer / Meshorer, Eran / Dekel, Benjamin

    Molecular therapy. Methods & clinical development

    2023  Volume 29, Page(s) 329–346

    Abstract: Upscaling of kidney epithelial cells is crucial for renal regenerative medicine. Nonetheless, the adult kidney lacks a distinct stem cell hierarchy, limiting the ability to long-term propagate clonal populations of primary cells that retain renal ... ...

    Abstract Upscaling of kidney epithelial cells is crucial for renal regenerative medicine. Nonetheless, the adult kidney lacks a distinct stem cell hierarchy, limiting the ability to long-term propagate clonal populations of primary cells that retain renal identity. Toward this goal, we tested the paradigm of shifting the balance between differentiation and stemness in the kidney by introducing a single pluripotency factor, OCT4. Here we show that ectopic expression of OCT4 in human adult kidney epithelial cells (hKEpC) induces the cells to dedifferentiate, stably proliferate, and clonally emerge over many generations. Control hKEpC dedifferentiate, assume fibroblastic morphology, and completely lose clonogenic capacity. Analysis of gene expression and histone methylation patterns revealed that OCT4 represses the HNF1B gene module, which is critical for kidney epithelial differentiation, and concomitantly activates stemness-related pathways. OCT4-hKEpC can be long-term expanded in the dedifferentiated state that is primed for renal differentiation. Thus, when expanded OCT4-hKEpC are grown as kidney spheroids (OCT4-kSPH), they reactivate the HNF1B gene signature, redifferentiate, and efficiently generate renal structures
    Language English
    Publishing date 2023-04-23
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2872938-9
    ISSN 2329-0501 ; 2329-0501
    ISSN (online) 2329-0501
    ISSN 2329-0501
    DOI 10.1016/j.omtm.2023.04.005
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Human kidney clonal proliferation disclose lineage-restricted precursor characteristics

    Cohen-Zontag Osnat / Gershon Rotem / Harari-Steinberg Orit / Kanter Itamar / Omer Dorit / Pleniceanu Oren / Tam Gal / Oriel Sarit / Ben Hur Herzel / Katz Guy / Zohar Dotan / Kalisky Tomer / Dekel Benjamin / Pode-Shakked Naomi

    Scientific Reports, Vol 10, Iss 1, Pp 1-

    2020  Volume 11

    Abstract: Abstract In-vivo single cell clonal analysis in the adult mouse kidney has previously shown lineage-restricted clonal proliferation within varying nephron segments as a mechanism responsible for cell replacement and local regeneration. To analyze ex-vivo ...

    Abstract Abstract In-vivo single cell clonal analysis in the adult mouse kidney has previously shown lineage-restricted clonal proliferation within varying nephron segments as a mechanism responsible for cell replacement and local regeneration. To analyze ex-vivo clonal growth, we now preformed limiting dilution to generate genuine clonal cultures from one single human renal epithelial cell, which can give rise to up to 3.4 * 106 cells, and analyzed their characteristics using transcriptomics. A comparison between clonal cultures revealed restriction to either proximal or distal kidney sub-lineages with distinct cellular and molecular characteristics; rapidly amplifying de-differentiated clones and a stably proliferating cuboidal epithelial-appearing clones, respectively. Furthermore, each showed distinct molecular features including cell-cycle, epithelial-mesenchymal transition, oxidative phosphorylation, BMP signaling pathway and cell surface markers. In addition, analysis of clonal versus bulk cultures show early clones to be more quiescent, with elevated expression of renal developmental genes and overall reduction in renal identity markers, but with an overlapping expression of nephron segment identifiers and multiple identity. Thus, ex-vivo clonal growth mimics the in-vivo situation displaying lineage-restricted precursor characteristics of mature renal cells. These data suggest that for reconstruction of varying renal lineages with human adult kidney based organoid technology and kidney regeneration ex-vivo, use of multiple heterogeneous precursors is warranted.
    Keywords Medicine ; R ; Science ; Q
    Subject code 616
    Language English
    Publishing date 2020-12-01T00:00:00Z
    Publisher Nature Portfolio
    Document type Article ; Online
    Database BASE - Bielefeld Academic Search Engine (life sciences selection)

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  3. Article ; Online: Author Correction: Human kidney clonal proliferation disclose lineage-restricted precursor characteristics.

    Cohen-Zontag, Osnat / Gershon, Rotem / Harari-Steinberg, Orit / Kanter, Itamar / Omer, Dorit / Pleniceanu, Oren / Tam, Gal / Oriel, Sarit / Ben-Hur, Herzl / Katz, Guy / Dotan, Zohar / Kalisky, Tomer / Dekel, Benjamin / Pode-Shakked, Naomi

    Scientific reports

    2021  Volume 11, Issue 1, Page(s) 6970

    Language English
    Publishing date 2021-03-22
    Publishing country England
    Document type Published Erratum
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-021-86157-7
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article ; Online: Human kidney clonal proliferation disclose lineage-restricted precursor characteristics.

    Cohen-Zontag, Osnat / Gershon, Rotem / Harari-Steinberg, Orit / Kanter, Itamar / Omer, Dorit / Pleniceanu, Oren / Tam, Gal / Oriel, Sarit / Ben-Hur, Herzel / Katz, Guy / Dotan, Zohar / Kalisky, Tomer / Dekel, Benjamin / Pode-Shakked, Naomi

    Scientific reports

    2020  Volume 10, Issue 1, Page(s) 22097

    Abstract: In-vivo single cell clonal analysis in the adult mouse kidney has previously shown lineage-restricted clonal proliferation within varying nephron segments as a mechanism responsible for cell replacement and local regeneration. To analyze ex-vivo clonal ... ...

    Abstract In-vivo single cell clonal analysis in the adult mouse kidney has previously shown lineage-restricted clonal proliferation within varying nephron segments as a mechanism responsible for cell replacement and local regeneration. To analyze ex-vivo clonal growth, we now preformed limiting dilution to generate genuine clonal cultures from one single human renal epithelial cell, which can give rise to up to 3.4 * 10
    MeSH term(s) Cell Differentiation/genetics ; Cell Proliferation/genetics ; Clonal Evolution/genetics ; Computational Biology ; Epithelial Cells/cytology ; Epithelial-Mesenchymal Transition/genetics ; Humans ; Kidney/cytology ; Kidney/growth & development ; Mesoderm/growth & development ; Mesoderm/metabolism ; Nephrons/growth & development ; Nephrons/metabolism ; Primary Cell Culture ; Regeneration/genetics ; Single-Cell Analysis ; Stem Cells/cytology
    Language English
    Publishing date 2020-12-16
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/s41598-020-78366-3
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Evidence of In Vitro Preservation of Human Nephrogenesis at the Single-Cell Level.

    Pode-Shakked, Naomi / Gershon, Rotem / Tam, Gal / Omer, Dorit / Gnatek, Yehudit / Kanter, Itamar / Oriel, Sarit / Katz, Guy / Harari-Steinberg, Orit / Kalisky, Tomer / Dekel, Benjamin

    Stem cell reports

    2017  Volume 9, Issue 1, Page(s) 279–291

    Abstract: During nephrogenesis, stem/progenitor cells differentiate and give rise to early nephron structures that segment to proximal and distal nephron cell types. Previously, we prospectively isolated progenitors from human fetal kidney (hFK) utilizing a ... ...

    Abstract During nephrogenesis, stem/progenitor cells differentiate and give rise to early nephron structures that segment to proximal and distal nephron cell types. Previously, we prospectively isolated progenitors from human fetal kidney (hFK) utilizing a combination of surface markers. However, upon culture nephron progenitors differentiated and could not be robustly maintained in vitro. Here, by culturing hFK in a modified medium used for in vitro growth of mouse nephron progenitors, and by dissection of NCAM
    Language English
    Publishing date 2017-07-11
    Publishing country United States
    Document type Journal Article
    ZDB-ID 2720528-9
    ISSN 2213-6711 ; 2213-6711
    ISSN (online) 2213-6711
    ISSN 2213-6711
    DOI 10.1016/j.stemcr.2017.04.026
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Dissecting Stages of Human Kidney Development and Tumorigenesis with Surface Markers Affords Simple Prospective Purification of Nephron Stem Cells.

    Pode-Shakked, Naomi / Pleniceanu, Oren / Gershon, Rotem / Shukrun, Rachel / Kanter, Itamar / Bucris, Efrat / Pode-Shakked, Ben / Tam, Gal / Tam, Hadar / Caspi, Revital / Pri-Chen, Sara / Vax, Einav / Katz, Guy / Omer, Dorit / Harari-Steinberg, Orit / Kalisky, Tomer / Dekel, Benjamin

    Scientific reports

    2016  Volume 6, Page(s) 23562

    Abstract: When assembling a nephron during development a multipotent stem cell pool becomes restricted as differentiation ensues. A faulty differentiation arrest in this process leads to transformation and initiation of a Wilms' tumor. Mapping these transitions ... ...

    Abstract When assembling a nephron during development a multipotent stem cell pool becomes restricted as differentiation ensues. A faulty differentiation arrest in this process leads to transformation and initiation of a Wilms' tumor. Mapping these transitions with respective surface markers affords accessibility to specific cell subpopulations. NCAM1 and CD133 have been previously suggested to mark human renal progenitor populations. Herein, using cell sorting, RNA sequencing, in vitro studies with serum-free media and in vivo xenotransplantation we demonstrate a sequential map that links human kidney development and tumorigenesis; In nephrogenesis, NCAM1(+)CD133(-) marks SIX2(+) multipotent renal stem cells transiting to NCAM1(+)CD133(+) differentiating segment-specific SIX2(-) epithelial progenitors and NCAM1(-)CD133(+) differentiated nephron cells. In tumorigenesis, NCAM1(+)CD133(-) marks SIX2(+) blastema that includes the ALDH1(+) WT cancer stem/initiating cells, while NCAM1(+)CD133(+) and NCAM1(-)CD133(+) specifying early and late epithelial differentiation, are severely restricted in tumor initiation capacity and tumor self-renewal. Thus, negative selection for CD133 is required for defining NCAM1(+) nephron stem cells in normal and malignant nephrogenesis.
    MeSH term(s) AC133 Antigen/genetics ; AC133 Antigen/metabolism ; Animals ; Biomarkers/metabolism ; CD56 Antigen/genetics ; CD56 Antigen/metabolism ; Carcinogenesis/genetics ; Carcinogenesis/metabolism ; Cells, Cultured ; Child ; Child, Preschool ; Female ; Gene Expression Regulation ; Homeodomain Proteins/genetics ; Homeodomain Proteins/metabolism ; Humans ; Immunohistochemistry ; Infant ; Kidney/embryology ; Kidney/metabolism ; Male ; Mice, Inbred NOD ; Neoplastic Stem Cells/metabolism ; Neoplastic Stem Cells/pathology ; Nephrons/cytology ; Nephrons/metabolism ; Nerve Tissue Proteins/genetics ; Nerve Tissue Proteins/metabolism ; Organogenesis/genetics ; Prospective Studies ; Reverse Transcriptase Polymerase Chain Reaction ; Stem Cells/metabolism ; Transplantation, Heterologous ; Tumor Cells, Cultured
    Chemical Substances AC133 Antigen ; Biomarkers ; CD56 Antigen ; Homeodomain Proteins ; NCAM1 protein, human ; Nerve Tissue Proteins ; SIX2 protein, human
    Language English
    Publishing date 2016-03-29
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2615211-3
    ISSN 2045-2322 ; 2045-2322
    ISSN (online) 2045-2322
    ISSN 2045-2322
    DOI 10.1038/srep23562
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