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  1. Article ; Online: Generation of two human NRF2 knockout iPSC clones using CRISPR/Cas9 editing.

    Merkert, Sylvia / Haase, Alexandra / Dahlmann, Julia / Göhring, Gudrun / Waqas, Fakhar H / Pessler, Frank / Martin, Ulrich / Olmer, Ruth

    Stem cell research

    2023  Volume 69, Page(s) 103090

    Abstract: The nuclear factor erythroid 2-related factor 2 (NFE2L2, known as NRF2) regulates the expression of antioxidative and anti-inflammatory proteins. In order to investigate its impact during viral infections and testing of antiviral compounds, we applied ... ...

    Abstract The nuclear factor erythroid 2-related factor 2 (NFE2L2, known as NRF2) regulates the expression of antioxidative and anti-inflammatory proteins. In order to investigate its impact during viral infections and testing of antiviral compounds, we applied CRISPR/Cas9 editing to eliminate NRF2 in the human iPS cell line MHHi001-A and generated two NRF2 knockout iPSC clones MHHi001-A-6 and MHHi001-A-7. After differentiation into epithelia or endothelial cells, these cells are useful tools to examine the antiviral effects of activators of the NRF2 signaling pathway.
    MeSH term(s) Humans ; CRISPR-Cas Systems/genetics ; Induced Pluripotent Stem Cells/metabolism ; NF-E2-Related Factor 2/genetics ; NF-E2-Related Factor 2/metabolism ; Endothelial Cells/metabolism ; Clone Cells/metabolism
    Chemical Substances NF-E2-Related Factor 2
    Language English
    Publishing date 2023-04-01
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2393143-7
    ISSN 1876-7753 ; 1873-5061
    ISSN (online) 1876-7753
    ISSN 1873-5061
    DOI 10.1016/j.scr.2023.103090
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article ; Online: Standardized production of hPSC-derived cardiomyocyte aggregates in stirred spinner flasks.

    Kriedemann, Nils / Triebert, Wiebke / Teske, Jana / Mertens, Mira / Franke, Annika / Ullmann, Kevin / Manstein, Felix / Drakhlis, Lika / Haase, Alexandra / Halloin, Caroline / Martin, Ulrich / Zweigerdt, Robert

    Nature protocols

    2024  

    Abstract: A promising cell-therapy approach for heart failure aims at differentiating human pluripotent stem cells (hPSCs) into functional cardiomyocytes (CMs) in vitro to replace the disease-induced loss of patients' heart muscle cells in vivo. But many ... ...

    Abstract A promising cell-therapy approach for heart failure aims at differentiating human pluripotent stem cells (hPSCs) into functional cardiomyocytes (CMs) in vitro to replace the disease-induced loss of patients' heart muscle cells in vivo. But many challenges remain for the routine clinical application of hPSC-derived CMs (hPSC-CMs), including good manufacturing practice (GMP)-compliant production strategies. This protocol describes the efficient generation of hPSC-CM aggregates in suspension culture, emphasizing process simplicity, robustness and GMP compliance. The strategy promotes clinical translation and other applications that require large numbers of CMs. Using a simple spinner-flask platform, this protocol is applicable to a broad range of users with general experience in handling hPSCs without extensive know-how in biotechnology. hPSCs are expanded in monolayer to generate the required cell numbers for process inoculation in suspension culture, followed by stirring-controlled formation of cell-only aggregates at a 300-ml scale. After 48 h at checkpoint (CP) 0, chemically defined cardiac differentiation is induced by WNT-pathway modulation through use of the glycogen-synthase kinase-3 inhibitor CHIR99021 (WNT agonist), which is replaced 24 h later by the chemical WNT-pathway inhibitor IWP-2. The exact application of the described process parameters is important to ensure process efficiency and robustness. After 10 d of differentiation (CP I), the production of ≥100 × 10
    Language English
    Publishing date 2024-03-28
    Publishing country England
    Document type Journal Article ; Review
    ZDB-ID 2244966-8
    ISSN 1750-2799 ; 1754-2189
    ISSN (online) 1750-2799
    ISSN 1754-2189
    DOI 10.1038/s41596-024-00976-2
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Matrix-free human pluripotent stem cell manufacturing by seed train approach and intermediate cryopreservation.

    Ullmann, Kevin / Manstein, Felix / Triebert, Wiebke / Kriedemann, Nils / Franke, Annika / Teske, Jana / Mertens, Mira / Lupanow, Victoria / Göhring, Gudrun / Haase, Alexandra / Martin, Ulrich / Zweigerdt, Robert

    Stem cell research & therapy

    2024  Volume 15, Issue 1, Page(s) 89

    Abstract: Background: Human pluripotent stem cells (hPSCs) have an enormous therapeutic potential, but large quantities of cells will need to be supplied by reliable, economically viable production processes. The suspension culture (three-dimensional; 3D) of ... ...

    Abstract Background: Human pluripotent stem cells (hPSCs) have an enormous therapeutic potential, but large quantities of cells will need to be supplied by reliable, economically viable production processes. The suspension culture (three-dimensional; 3D) of hPSCs in stirred tank bioreactors (STBRs) has enormous potential for fuelling these cell demands. In this study, the efficient long-term matrix-free suspension culture of hPSC aggregates is shown.
    Methods and results: STBR-controlled, chemical aggregate dissociation and optimized passage duration of 3 or 4 days promotes exponential hPSC proliferation, process efficiency and upscaling by a seed train approach. Intermediate high-density cryopreservation of suspension-derived hPSCs followed by direct STBR inoculation enabled complete omission of matrix-dependent 2D (two-dimensional) culture. Optimized 3D cultivation over 8 passages (32 days) cumulatively yielded ≈4.7 × 10
    Conclusions: Together, an entirely matrix-free, highly efficient, flexible and automation-friendly hPSC expansion strategy is demonstrated, facilitating the development of good manufacturing practice-compliant closed-system manufacturing in large scale.
    MeSH term(s) Humans ; Cell Culture Techniques/methods ; Pluripotent Stem Cells/metabolism ; Cell Differentiation ; Bioreactors ; Cryopreservation
    Language English
    Publishing date 2024-03-25
    Publishing country England
    Document type Journal Article
    ZDB-ID 2548671-8
    ISSN 1757-6512 ; 1757-6512
    ISSN (online) 1757-6512
    ISSN 1757-6512
    DOI 10.1186/s13287-024-03699-z
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article: Initial WNT/β-Catenin Activation Enhanced Mesoderm Commitment, Extracellular Matrix Expression, Cell Aggregation and Cartilage Tissue Yield From Induced Pluripotent Stem Cells.

    Kreuser, Ursula / Buchert, Justyna / Haase, Alexandra / Richter, Wiltrud / Diederichs, Solvig

    Frontiers in cell and developmental biology

    2020  Volume 8, Page(s) 581331

    Abstract: Mesodermal differentiation of induced pluripotent stem cells (iPSCs) ...

    Abstract Mesodermal differentiation of induced pluripotent stem cells (iPSCs)
    Language English
    Publishing date 2020-10-30
    Publishing country Switzerland
    Document type Journal Article
    ZDB-ID 2737824-X
    ISSN 2296-634X
    ISSN 2296-634X
    DOI 10.3389/fcell.2020.581331
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  5. Article ; Online: Generation of human induced pluripotent stem cell lines encoding for genetically encoded calcium indicators RCaMP1h and GCaMP6f.

    Ricci Signorini, Maria Elena / Szepes, Monika / Melchert, Anna / Bakar, Mine / Merkert, Sylvia / Haase, Alexandra / Göhring, Gudrun / Martin, Ulrich / Gruh, Ina

    Stem cell research

    2022  Volume 60, Page(s) 102697

    Abstract: Calcium plays a key role in cardiomyocytes (CMs) for the translation of the electrical impulse of an action potential into contraction forces. A rapid, not-invasive fluorescence imaging technology allows for the monitoring of calcium transients in human ... ...

    Abstract Calcium plays a key role in cardiomyocytes (CMs) for the translation of the electrical impulse of an action potential into contraction forces. A rapid, not-invasive fluorescence imaging technology allows for the monitoring of calcium transients in human induced pluripotent stem cell derived-cardiomyocytes (hiPSC-CMs) to investigate the cardiac electrophysiology in vitro and after cell transplantation in vivo. The genetically encoded calcium indicators (GECIs) GCaMP6f or RCaMP1h were successfully transfected in the previously established hiPSC line MHHi001-A, together with a cardiac specific antibiotic selection cassette facilitating the monitoring of the calcium handling in highly pure populations of hiPSC-CMs.
    MeSH term(s) Action Potentials ; Calcium/metabolism ; Cell Differentiation ; Humans ; Induced Pluripotent Stem Cells/metabolism ; Myocytes, Cardiac/metabolism
    Chemical Substances Calcium (SY7Q814VUP)
    Language English
    Publishing date 2022-02-02
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2393143-7
    ISSN 1876-7753 ; 1873-5061
    ISSN (online) 1876-7753
    ISSN 1873-5061
    DOI 10.1016/j.scr.2022.102697
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article ; Online: Generation of non-transgenic iPS cells from human cord blood CD34

    Haase, Alexandra / Göhring, Gudrun / Martin, Ulrich

    Stem cell research

    2017  Volume 21, Page(s) 71–73

    Abstract: Recently, many hurdles and limitations for production of clinically applicable iPSC derivatives have been overcome. Transgene-free iPSCs can be efficiently derived from easily accessible cell sources such as blood. Here we describe the generation of ... ...

    Abstract Recently, many hurdles and limitations for production of clinically applicable iPSC derivatives have been overcome. Transgene-free iPSCs can be efficiently derived from easily accessible cell sources such as blood. Here we describe the generation of transgene-free hiPS cells from cord blood derived CD34
    Language English
    Publishing date 2017-05
    Publishing country England
    Document type Journal Article
    ISSN 1876-7753
    ISSN (online) 1876-7753
    DOI 10.1016/j.scr.2017.03.022
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  7. Article ; Online: Generation of an induced pluripotent stem cell line, ZIPi021-A, from fibroblasts of a Prader-Willi syndrome patient with maternal uniparental disomy (mUPD).

    Heseding, Hannah / Jahn, Kirsten / Brändl, Björn / Haase, Alexandra / Shum, Ian O / Kohrn, Tim / Bleich, Stefan / Frieling, Helge / Martin, Ulrich / Müller, Franz-Josef / Wunderlich, Stephanie / Deest, Maximilian

    Stem cell research

    2023  Volume 71, Page(s) 103143

    Abstract: Prader-Willi syndrome (PWS) is a neurodevelopmental disorder caused by loss of paternal expression of imprinted genes on chromosome 15q11-q13. We established a human induced pluripotent stem cell line (hiPSC), ZIPi021-A, from fibroblasts of a 4-year-old ... ...

    Abstract Prader-Willi syndrome (PWS) is a neurodevelopmental disorder caused by loss of paternal expression of imprinted genes on chromosome 15q11-q13. We established a human induced pluripotent stem cell line (hiPSC), ZIPi021-A, from fibroblasts of a 4-year-old female PWS patient with the subtype of maternal uniparental disomy (mUPD). The generated hiPSC line was transgene-free, expressed pluripotency markers and showed the ability to differentiate into all three germ layers in vitro. The ZIPi021-A hiPSC line could be used as a cellular model for PWS in humans.
    MeSH term(s) Female ; Humans ; Child, Preschool ; Prader-Willi Syndrome/genetics ; Uniparental Disomy/genetics ; Induced Pluripotent Stem Cells/metabolism ; Neurodevelopmental Disorders ; Fibroblasts/metabolism ; Chromosomes, Human, Pair 15/genetics
    Language English
    Publishing date 2023-06-14
    Publishing country England
    Document type Case Reports ; Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2393143-7
    ISSN 1876-7753 ; 1873-5061
    ISSN (online) 1876-7753
    ISSN 1873-5061
    DOI 10.1016/j.scr.2023.103143
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  8. Book ; Online ; Thesis: Herstellung von humanen induzierten pluripotenten Stammzellen (iPS-Zellen) aus Nabelschnurblut

    Haase, Alexandra

    2010  

    Abstract: Induzierte pluripotente Stammzellen, Reprogrammierung, Nabelschnurblut. - Induced pluripotent stem cells, reprogramming, cord ... ...

    Author's details von Alexandra Haase
    Abstract Induzierte pluripotente Stammzellen, Reprogrammierung, Nabelschnurblut. - Induced pluripotent stem cells, reprogramming, cord blood
    Language German
    Size Online-Ressource (125 S., 7451 KB)
    Publisher Technische Informationsbibliothek u. Universitätsbibliothek
    Publishing place Hannover
    Document type Book ; Online ; Thesis
    Thesis / German Habilitation thesis Univ., Diss.--Hannover, 2011
    Database Library catalogue of the German National Library of Science and Technology (TIB), Hannover

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  9. Book ; Thesis: Herstellung von humanen induzierten pluripotenten Stammzellen (iPS-Zellen) aus Nabelschnurblut

    Haase, Alexandra

    2010  

    Abstract: Induzierte pluripotente Stammzellen, Reprogrammierung, Nabelschnurblut. - Induced pluripotent stem cells, reprogramming, cord ... ...

    Author's details von Alexandra Haase
    Abstract Induzierte pluripotente Stammzellen, Reprogrammierung, Nabelschnurblut. - Induced pluripotent stem cells, reprogramming, cord blood
    Language German
    Size III, II Bl., Bl. 12-124, Ill., graph. Darst.
    Document type Book ; Thesis
    Thesis / German Habilitation thesis Univ., Diss.--Hannover, 2011
    Database Library catalogue of the German National Library of Science and Technology (TIB), Hannover

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  10. Article ; Online: Generation of pulmonary arterial hypertension patient-specific induced pluripotent stem cell lines from three unrelated patients with a heterozygous missense mutation in exon 12, a heterozygous in-frame deletion in exon 3 and a missense mutation in exon 11 of the BMPR2 gene.

    Usman, Abdulai / Haase, Alexandra / Merkert, Sylvia / Göhring, Gudrun / Hansmann, Georg / Gall, Henning / Schermuly, Ralph / Martin, Ulrich / Olmer, Ruth

    Stem cell research

    2021  Volume 55, Page(s) 102488

    Abstract: Loss-of-function mutations in the bone morphogenetic protein receptor 2 (BMPR2) gene are common in heritable or idiopathic pulmonary arterial hypertension (PAH), and can result in functional impairment of both endothelial and vascular smooth muscle cells. ...

    Abstract Loss-of-function mutations in the bone morphogenetic protein receptor 2 (BMPR2) gene are common in heritable or idiopathic pulmonary arterial hypertension (PAH), and can result in functional impairment of both endothelial and vascular smooth muscle cells. Here, we report 3 PAH patient-specific induced pluripotent stem cells (iPSC) lines from 3 unrelated patients harbouring different mutations in the BMPR2 gene: a heterozygous missense mutation in exon 12, a heterozygous frame shift deletion in exon 3, and a heterozygous missense mutation in exon 11. These cell lines will serve as a valuable resource to model PAH in vitro.
    MeSH term(s) Bone Morphogenetic Protein Receptors, Type II/genetics ; Exons/genetics ; Familial Primary Pulmonary Hypertension ; Humans ; Hypertension, Pulmonary/genetics ; Induced Pluripotent Stem Cells ; Mutation ; Mutation, Missense/genetics ; Pulmonary Arterial Hypertension
    Chemical Substances BMPR2 protein, human (EC 2.7.11.30) ; Bone Morphogenetic Protein Receptors, Type II (EC 2.7.11.30)
    Language English
    Publishing date 2021-08-05
    Publishing country England
    Document type Journal Article ; Research Support, Non-U.S. Gov't
    ZDB-ID 2393143-7
    ISSN 1876-7753 ; 1873-5061
    ISSN (online) 1876-7753
    ISSN 1873-5061
    DOI 10.1016/j.scr.2021.102488
    Database MEDical Literature Analysis and Retrieval System OnLINE

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