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  • 1
    Online Resource
    Online Resource
    Elsevier BV ; 2014
    In:  The American Journal of Pathology Vol. 184, No. 3 ( 2014-03), p. 827-841
    In: The American Journal of Pathology, Elsevier BV, Vol. 184, No. 3 ( 2014-03), p. 827-841
    Type of Medium: Online Resource
    ISSN: 0002-9440
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    Language: English
    Publisher: Elsevier BV
    Publication Date: 2014
    detail.hit.zdb_id: 1480207-7
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  • 2
    Online Resource
    Online Resource
    Wiley ; 2021
    In:  Annals of the New York Academy of Sciences Vol. 1487, No. 1 ( 2021-03), p. 12-20
    In: Annals of the New York Academy of Sciences, Wiley, Vol. 1487, No. 1 ( 2021-03), p. 12-20
    Abstract: Organ and tissue repair are complex processes involving signaling molecules, growth factors, and cell cycle regulators that act in concert to promote cell division and differentiation at sites of injury. In embryonic development, progenitor fetal cells are actively involved in reparative mechanisms and display a biphasic interaction with the mother; and there is constant trafficking of fetal cells into maternal circulation and vice versa. This phenomenon of fetal microchimerism may have significant impact considering the primitive, multilineage nature of these cells. In published work, we have reported that fetal‐derived placental cells expressing the homeodomain protein CDX2 retain all “stem” functional proteins of embryonic stem cells yet are endowed with additional functions in areas of growth, survival, homing, and immune modulation. These cells exhibit multipotency in vitro and in vivo , giving rise to spontaneously beating cardiomyocytes and vascular cells. In mouse models, CDX2 cells from female placentas can be administered intravenously to male mice subjected to myocardial infarction with subsequent homing of the CDX2 cells to infarcted areas and evidence of cellular regeneration with enhanced cardiac function. Elucidating the role of microchimeric fetal‐derived placental cells may have broader scientific potential, as one can envision allogeneic cell therapy strategies targeted at tissue regeneration for a variety of organ systems.
    Type of Medium: Online Resource
    ISSN: 0077-8923 , 1749-6632
    URL: Issue
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    Language: English
    Publisher: Wiley
    Publication Date: 2021
    detail.hit.zdb_id: 2834079-6
    detail.hit.zdb_id: 211003-9
    detail.hit.zdb_id: 2071584-5
    SSG: 11
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  • 3
    Online Resource
    Online Resource
    Elsevier BV ; 2022
    In:  JACC: Basic to Translational Science Vol. 7, No. 1 ( 2022-01), p. 67-69
    In: JACC: Basic to Translational Science, Elsevier BV, Vol. 7, No. 1 ( 2022-01), p. 67-69
    Type of Medium: Online Resource
    ISSN: 2452-302X
    Language: English
    Publisher: Elsevier BV
    Publication Date: 2022
    detail.hit.zdb_id: 2865010-4
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  • 4
    Online Resource
    Online Resource
    Ovid Technologies (Wolters Kluwer Health) ; 2019
    In:  Circulation Research Vol. 125, No. Suppl_1 ( 2019-08-02)
    In: Circulation Research, Ovid Technologies (Wolters Kluwer Health), Vol. 125, No. Suppl_1 ( 2019-08-02)
    Abstract: The limited regeneration of adult mammalian heart has prompted the need to recognize novel strategies that can restore contractile function in heart disease. However, in cell-based therapies the lack of an appropriate cell-type that can differentiate to cardiomyocytes in vivo persists as an ultimate unmet need. Our prior study demonstrates that experimental myocardial injury in pregnant mice triggers the flux of fetal cells via the maternal circulation into the injured heart where they undergo differentiation into diverse cardiac cell fates. Among those fetal cells, the expression of Caudal type homeobox2 (Cdx2); a trophoblast stem cell marker was unique. To understand the intriguing role of placental Cdx2 cells in cardiomyogenesis, we utilized a lineage-tracing strategy to label fetal-derived Cdx2 cells with enhanced green fluorescent protein (Cdx2-eGFP). Cdx2-eGFP cells were characterized and assayed for cardiac differentiation in vitro and in vivo using a mouse model of myocardial infarction. Cdx2-eGFP cells clonally proliferated and differentiated into spontaneously beating cardiomyocytes and vascular cells in vitro , signifying a multipotent nature compared to the Cdx2 negative cell population. When administered via tail vein to infarcted wild-type male mice, Cdx2-eGFP cells selectively and robustly homed to the injured heart and differentiated to cardiomyocytes and blood vessels, significantly improving the contractility noted by magnetic resonance imaging. Proteomics and immune transcriptomics studies of Cdx2-eGFP cells compared to embryonic stem (ES) cells reveal that they appear to retain ‘stem’-related functions of ES cells, but exhibit unique signatures for homing and survival in addition to being immunologically naive. Blocking CXCR4, during the migration of Cdx2-eGFP cells to SDF1α suggested a possible role for SDF1-CXCR4 signaling in the mechanistic basis of homing. Advancing towards a translational role, we demonstrate that CDX2 expressing cells can be isolated from the chorionic region of human term placenta. Our results herein may represent a paradigmatic shift in the way we approach early embryonic lineages and cell fate choices and will establish the translational potential of placental Cdx2 cells for cardiac repair.
    Type of Medium: Online Resource
    ISSN: 0009-7330 , 1524-4571
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    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2019
    detail.hit.zdb_id: 1467838-X
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  • 5
    In: Circulation, Ovid Technologies (Wolters Kluwer Health), Vol. 146, No. Suppl_1 ( 2022-11-08)
    Abstract: Regenerative biology hinges on exploration of evolutionarily preserved healing mechanisms to maintain and restore homeostasis. CDX2 is one such conserved factor that regulates trophoblast stem cell differentiation in the placenta. We previously demonstrated that murine placental Cdx2 lineage cells are multipotent and restored cardiac function even in male mice after myocardial infarction. We have now isolated CDX2 cells from over 35 patients’ placentas and studied their properties. Using huCDX2 promoter-driven mCherry lentivirus, CDX2 cells were isolated and multi-omics analyses were carried out versus H9ES cells. In vitro cardiac and vascular differentiation was examined. CDX2 cells expressed HLAG (50.5%±5) and cytokeratin7 (95%±5) supporting a trophoblast progenitor identity. Bulk RNA-seq showed 1065 upregulated genes. Differentially expressed genes like NKX2.5, GATA4 , and TBX5 attribute a cardiac committed progenitor feature to CDX2 cells. CDX2 cells downregulated pluripotency markers negating concerns of any undesirable proliferation. Whole-cell proteome of CDX2 cells revealed 252 unique proteins of which 73 were upregulated involving development and immune modulation including FGF, EFGR, and the Map kinase/ERK pathway crucial for cardiac development, in addition to unique cell surface proteins, which can be utilized in place of lentivirus for live-cell selection. When plated on neonatal murine cardiomyocyte feeders, CDX2 cells robustly differentiated into beating cardiomyocytes expressing cTnT and alpha-actinin. Using an alternate feeder-free method, we observed 80-90% cTnT expression in differentiated CDX2 cardiomyocytes. Additionally, CDX2 cells gave rise to endothelial lineage cells expressing vWF. CDX2 cells further formed tube-like structures on matrigel-based endothelial assays and demonstrated uptake of acetylated LDL suggesting functional endothelial generation. We further demonstrated that CDX2 cells can be passaged and significantly expanded ex vivo (50,000 to 1million cells), while retaining cardiovascular potential. These results bring us closer to translation as placenta-derived CDX2 cells may form the basis for a novel clinical approach to cardiovascular regeneration.
    Type of Medium: Online Resource
    ISSN: 0009-7322 , 1524-4539
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2022
    detail.hit.zdb_id: 1466401-X
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  • 6
    In: Circulation Research, Ovid Technologies (Wolters Kluwer Health), Vol. 121, No. suppl_1 ( 2017-07-21)
    Abstract: Stem cell-based therapies for cardiac regeneration are of crucial importance and an ideal cell-type is yet to be established. We previously reported that fetal cells from placenta “home” to injured maternal heart and approximately 40% (40/100) of the migrating cells expressed homeodomain protein Cdx2. This interesting observation led us to hypothesize that placental Cdx2 could be a novel cell target for cardiac differentiation. To understand this phenomenon, we employed a cre-lox strategy that labeled Cdx2 cells in placenta with e-GFP and induced myocardial infarction (MI) in pregnant mice at mid-gestation. The maternal heart was analyzed 4 weeks post-MI for the presence of Cdx2-eGFP-derived cardiomyocytes. Additionally, Cdx2 cells were isolated from late-gestation placenta and assayed for cardiac differentiation in vitro followed by live cell imaging. Phenotypic and whole-cell proteomic analysis, clonal and vascular lineage differentiation and immune profiling were carried out subsequently. We observed that Cdx2 cells migrated to injured maternal hearts and differentiated into cardiomyocytes highlighting the functional significance of fetal-maternal stem cell transfer. Additionally, isolated Cdx2 cells from the late placenta differentiated into spontaneously beating cardiomyocytes and expressed structural proteins cardiac troponin T(cTnT), α-sarcomeric actinin and gap junction protein Cx43. These cells underwent clonal expansion and differentiated into endothelial and smooth muscle lineages in culture indicative of their multipotent nature. Low expression of MHC molecules and other components of the immune-response, infer that these cells possess the ability to evade host immune surveillance. Proteomic analysis demonstrated that 145 proteins were uniquely identified in the Cdx2 cells compared to embryonic stem cells. These protein networks reflected an increased activation of functions involving migration, fertility, homing, and chemotaxis. Our study is the first to demonstrate that Cdx2 may play a role in cardiac differentiation and delineate multipotent cells in placenta with an inherent “homing” ability. These findings point to a potential role for Cdx2 cells in cardiac regenerative therapies using allogeneic cells.
    Type of Medium: Online Resource
    ISSN: 0009-7330 , 1524-4571
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    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2017
    detail.hit.zdb_id: 1467838-X
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  • 7
    Online Resource
    Online Resource
    Ovid Technologies (Wolters Kluwer Health) ; 2020
    In:  Circulation Research Vol. 127, No. Suppl_1 ( 2020-07-31)
    In: Circulation Research, Ovid Technologies (Wolters Kluwer Health), Vol. 127, No. Suppl_1 ( 2020-07-31)
    Abstract: Coronary artery disease and heart failure are among the leading causes of mortality in United States and worldwide. The limited proliferation of the adult mammalian heart has prompted the need to recognize novel strategies that can restore contractile function in heart disease. We recently reported that cells expressing trophoblast marker Cdx2 isolated from murine end-gestation placentas are multipotent and immune-privileged, with homing ability to travel to sites of cardiac injury to regenerate injured myocardium. Since murine Cdx2 cells are immunologically naïve and trophoblast-mediated placentation is a conserved phenomenon, our findings are ripe for translational application. Here we demonstrate the expression of CDX2 and isolation of CDX2 cells from human term placentas highlighting a potentially critical cell source for allogeneic therapy for cardiac regeneration. We studied de-identified human term placenta tissues from three different anatomical sites from three different patients. Formalin-fixed paraffin embedded samples, second trimester chorionic villus samples and de-identified fresh chorion samples were utilized. Using a multiparametric approach including transcript analysis and Sanger sequencing, immunoblot and immunofluorescence analysis, with the subsequent screening of different anatomical sites, we observed that CDX2 is present in the chorionic (fetal cytotrophoblast) portion of human term placentas including the chorionic villous samples (CVS). Subsequent validation on positive control cells DLD1 (human colon carcinoma cell line), versus HeLa cells (low endogenous CDX2 expression) was performed. Visualization of CDX2 expression showed the nuclear localization in the villi region. We have further demonstrated using a CDX2 promoter-driven fluorescence-based lentiviral expression system, we can isolate viable CDX2 cells from fresh term placenta. We demonstrate for the first time that CDX2 cells are present and can be isolated from term human placentas for prospective cardiomyocgenic differentiation. A step closer to the translational approach, our results herein may represent a paradigmatic shift in the way we approach early embryonic lineages and cell fate choices towards the identification of an unexplored human cell-based approach for cardiac regeneration.
    Type of Medium: Online Resource
    ISSN: 0009-7330 , 1524-4571
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    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2020
    detail.hit.zdb_id: 1467838-X
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  • 8
    Online Resource
    Online Resource
    Ovid Technologies (Wolters Kluwer Health) ; 2017
    In:  Circulation Research Vol. 121, No. suppl_1 ( 2017-07-21)
    In: Circulation Research, Ovid Technologies (Wolters Kluwer Health), Vol. 121, No. suppl_1 ( 2017-07-21)
    Abstract: The adult mammalian heart is known to have a very low abundance of progenitor cells which can take part in active cycling and regeneration after damage. Cardiomyocytes exit the cell cycle soon after birth coincident with the silencing of cyclin A2 (CCNA2). In our previous studies, we demonstrated that viral delivery of Ccna2 induces cardiac regeneration in infarcted hearts of small and large animal models. However, the molecular mechanism whereby Ccna2 induces cardiac regeneration and increase in cardiac function deserves further study. To explore further, we isolated adult mouse cardiomyocytes and induced Ccna2 expression by using adenovirus transfection and cultured them for 3 weeks. Co-expression of the mature cardiac marker troponin Tc with the immature cardiac marker non-muscle myosin IIB was observed. Additionally, expression of epithelial to mesenchymal transition markers (vimentin and FSP1) was observed. Also, decreased expression of mature cardiac markers α-MHC , ckmt2 and cTnT was noted. To study the factors responsible for human cardiomyocyte plasticity and cell division, we have optimized a novel method for culturing adult human cardiomyocytes in our laboratory. We cultured cardiomyocytes isolated from heart tissue obtained from a 55 yr old male patient. After transfection with CCNA2 adenovirus made for human use (cTnT promoter driving human CCNA2 cDNA), they were co-transfected with two more adenoviruses (1) cTnT-GFP to label cardiomyocytes (green) and (2) CMV-α-actinin-m-Cherry to label the sarcomere (red). Time lapse live epifluorescence microscopy was carried out for 70 hrs and time lapse movies were prepared (please refer the youtube link to see a representative time lapse movie https://youtu.be/OBrJGCq7YCA ). Movies were analyzed to calculate the cytokinesis index in samples transfected with (Test) and without (Null) CCNA2 adenoviruses. We observed a significantly higher cytokinesis index in CCNA2 samples versus Null. We are further investigating the role of cyclin A2 in dedifferentiation of adult human cardiomyocytes to generate immature or progenitor cardiac cells and their contractile status, which could be utilized for regeneration and functional restoration of damaged adult heart tissue.
    Type of Medium: Online Resource
    ISSN: 0009-7330 , 1524-4571
    RVK:
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2017
    detail.hit.zdb_id: 1467838-X
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  • 9
    Online Resource
    Online Resource
    Ovid Technologies (Wolters Kluwer Health) ; 2018
    In:  Circulation Research Vol. 123, No. 1 ( 2018-06-22), p. 24-26
    In: Circulation Research, Ovid Technologies (Wolters Kluwer Health), Vol. 123, No. 1 ( 2018-06-22), p. 24-26
    Type of Medium: Online Resource
    ISSN: 0009-7330 , 1524-4571
    RVK:
    Language: English
    Publisher: Ovid Technologies (Wolters Kluwer Health)
    Publication Date: 2018
    detail.hit.zdb_id: 1467838-X
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  • 10
    Online Resource
    Online Resource
    Proceedings of the National Academy of Sciences ; 2019
    In:  Proceedings of the National Academy of Sciences Vol. 116, No. 24 ( 2019-06-11), p. 11786-11795
    In: Proceedings of the National Academy of Sciences, Proceedings of the National Academy of Sciences, Vol. 116, No. 24 ( 2019-06-11), p. 11786-11795
    Abstract: The extremely limited regenerative potential of adult mammalian hearts has prompted the need for novel cell-based therapies that can restore contractile function in heart disease. We have previously shown the regenerative potential of mixed fetal cells that were naturally found migrating to the injured maternal heart. Exploiting this intrinsic mechanism led to the current hypothesis that Caudal-type homeobox-2 (Cdx2) cells in placenta may represent a novel cell type for cardiac regeneration. Using a lineage-tracing strategy, we specifically labeled fetal-derived Cdx2 cells with enhanced green fluorescent protein (eGFP). Cdx2-eGFP cells from end-gestation placenta were assayed for cardiac differentiation in vitro and in vivo using a mouse model of myocardial infarction. We observed that these cells differentiated into spontaneously beating cardiomyocytes (CMs) and vascular cells in vitro, indicating multipotentiality. When administered via tail vein to infarcted wild-type male mice, they selectively and robustly homed to the heart and differentiated to CMs and blood vessels, resulting in significant improvement in contractility as noted by MRI. Proteomics and immune transcriptomics studies of Cdx2-eGFP cells compared with embryonic stem (ES) cells reveal that they appear to retain “stem”-related functions of ES cells but exhibit unique signatures supporting roles in homing and survival, with an ability to evade immune surveillance, which is critical for cell-based therapy. Cdx2-eGFP cells may potentially represent a therapeutic advance in allogeneic cell therapy for cardiac repair.
    Type of Medium: Online Resource
    ISSN: 0027-8424 , 1091-6490
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    Language: English
    Publisher: Proceedings of the National Academy of Sciences
    Publication Date: 2019
    detail.hit.zdb_id: 209104-5
    detail.hit.zdb_id: 1461794-8
    SSG: 11
    SSG: 12
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