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  1. Article ; Online: Induced pluripotent stem cell technology in bone biology.

    Kidwai, Fahad K / Canalis, Ernesto / Robey, Pamela G

    Bone

    2023  Volume 172, Page(s) 116760

    Abstract: Technologies on the development and differentiation of human induced pluripotent stem cells (hiPSCs) are rapidly improving, and have been applied to create cell types relevant to the bone field. Differentiation protocols to form bona fide bone-forming ... ...

    Abstract Technologies on the development and differentiation of human induced pluripotent stem cells (hiPSCs) are rapidly improving, and have been applied to create cell types relevant to the bone field. Differentiation protocols to form bona fide bone-forming cells from iPSCs are available, and can be used to probe details of differentiation and function in depth. When applied to iPSCs bearing disease-causing mutations, the pathogenetic mechanisms of diseases of the skeleton can be elucidated, along with the development of novel therapeutics. These cells can also be used for development of cell therapies for cell and tissue replacement.
    MeSH term(s) Humans ; Induced Pluripotent Stem Cells/metabolism ; Cell Differentiation/genetics ; Biology
    Language English
    Publishing date 2023-04-06
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural ; Research Support, N.I.H., Extramural
    ZDB-ID 632515-4
    ISSN 1873-2763 ; 8756-3282
    ISSN (online) 1873-2763
    ISSN 8756-3282
    DOI 10.1016/j.bone.2023.116760
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  2. Article: Metabolic Quadrivalency in RSeT Human Embryonic Stem Cells.

    Chen, Kevin G / Park, Kyeyoon / Maric, Dragan / Johnson, Kory R / Robey, Pamela G / Mallon, Barbara S

    bioRxiv : the preprint server for biology

    2024  

    Abstract: One of the most important properties of human embryonic stem cells (hESCs) is related to their pluripotent states. In our recent study, we identified a previously unrecognized pluripotent state induced by RSeT medium. This state makes primed hESCs ... ...

    Abstract One of the most important properties of human embryonic stem cells (hESCs) is related to their pluripotent states. In our recent study, we identified a previously unrecognized pluripotent state induced by RSeT medium. This state makes primed hESCs resistant to conversion to naïve pluripotent state. In this study, we have further characterized the metabolic features in these RSeT hESCs, including metabolic gene expression, metabolomic analysis, and various functional assays. The commonly reported metabolic modes include glycolysis or both glycolysis and oxidative phosphorylation (i.e., metabolic bivalency) in pluripotent stem cells. However, besides the presence of metabolic bivalency, RSeT hESCs exhibited a unique metabolome with additional fatty acid oxidation and imbalanced nucleotide metabolism. This metabolic quadrivalency is linked to hESC growth independent of oxygen tension and restricted capacity for naïve reprogramming in these cells. Thus, this study provides new insights into pluripotent state transitions and metabolic stress-associated hPSC growth
    Language English
    Publishing date 2024-02-22
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2024.02.21.581486
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  3. Article ; Online: Microtissue Culture Provides Clarity on the Relative Chondrogenic and Hypertrophic Response of Bone-Marrow-Derived Stromal Cells to TGF-β1, BMP-2, and GDF-5.

    Franco, Rose Ann G / McKenna, Eamonn / Shajib, Md Shafiullah / Guillesser, Bianca / Robey, Pamela G / Crawford, Ross W / Doran, Michael R / Futrega, Kathryn

    Cells

    2023  Volume 13, Issue 1

    Abstract: Chondrogenic induction of bone-marrow-derived stromal cells (BMSCs) is typically accomplished with medium supplemented with growth factors (GF) from the transforming growth factor-beta (TGF-β)/bone morphogenetic factor (BMP) superfamily. In a previous ... ...

    Abstract Chondrogenic induction of bone-marrow-derived stromal cells (BMSCs) is typically accomplished with medium supplemented with growth factors (GF) from the transforming growth factor-beta (TGF-β)/bone morphogenetic factor (BMP) superfamily. In a previous study, we demonstrated that brief (1-3 days) stimulation with TGF-β1 was sufficient to drive chondrogenesis and hypertrophy using small-diameter microtissues generated from 5000 BMSC each. This biology is obfuscated in typical large-diameter pellet cultures, which suffer radial heterogeneity. Here, we investigated if brief stimulation (2 days) of BMSC microtissues with BMP-2 (100 ng/mL) or growth/differentiation factor (GDF-5, 100 ng/mL) was also sufficient to induce chondrogenic differentiation, in a manner comparable to TGF-β1 (10 ng/mL). Like TGF-β1, BMP-2 and GDF-5 are reported to stimulate chondrogenic differentiation of BMSCs, but the effects of transient or brief use in culture have not been explored. Hypertrophy is an unwanted outcome in BMSC chondrogenic differentiation that renders engineered tissues unsuitable for use in clinical cartilage repair. Using three BMSC donors, we observed that all GFs facilitated chondrogenesis, although the efficiency and the necessary duration of stimulation differed. Microtissues treated with 2 days or 14 days of TGF-β1 were both superior at producing extracellular matrix and expression of chondrogenic gene markers compared to BMP-2 and GDF-5 with the same exposure times. Hypertrophic markers increased proportionally with chondrogenic differentiation, suggesting that these processes are intertwined for all three GFs. The rapid action, or "temporal potency", of these GFs to induce BMSC chondrogenesis was found to be as follows: TGF-β1 > BMP-2 > GDF-5. Whether briefly or continuously supplied in culture, TGF-β1 was the most potent GF for inducing chondrogenesis in BMSCs.
    MeSH term(s) Humans ; Transforming Growth Factor beta1/pharmacology ; Growth Differentiation Factor 5/pharmacology ; Bone Marrow ; Chondrogenesis ; Transforming Growth Factor beta ; Hypertrophy ; Mesenchymal Stem Cells
    Chemical Substances Transforming Growth Factor beta1 ; Growth Differentiation Factor 5 ; Transforming Growth Factor beta
    Language English
    Publishing date 2023-12-23
    Publishing country Switzerland
    Document type Journal Article ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
    ZDB-ID 2661518-6
    ISSN 2073-4409 ; 2073-4409
    ISSN (online) 2073-4409
    ISSN 2073-4409
    DOI 10.3390/cells13010037
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  4. Article: SP7

    Franco, Rose Ann G / McKenna, Eamonn / Robey, Pamela G / Crawford, Ross W / Doran, Michael R / Futrega, Kathryn

    Journal of tissue engineering

    2023  Volume 14, Page(s) 20417314231177136

    Abstract: For bone marrow stromal cells (BMSC) to be useful in cartilage repair their propensity for hypertrophic differentiation must be overcome. A single day of TGF-β1 stimulation activates intrinsic signaling cascades in BMSCs which subsequently drives both ... ...

    Abstract For bone marrow stromal cells (BMSC) to be useful in cartilage repair their propensity for hypertrophic differentiation must be overcome. A single day of TGF-β1 stimulation activates intrinsic signaling cascades in BMSCs which subsequently drives both chondrogenic and hypertrophic differentiation. TGF-β1 stimulation upregulates
    Language English
    Publishing date 2023-06-21
    Publishing country England
    Document type Journal Article
    ZDB-ID 2573915-3
    ISSN 2041-7314
    ISSN 2041-7314
    DOI 10.1177/20417314231177136
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  5. Article: Transcriptomic signature and pro-osteoclastic secreted factors of abnormal bone marrow stromal cells in fibrous dysplasia.

    Michel, Zachary / Raborn, Layne N / Spencer, Tiahna / Pan, Kristen / Martin, Daniel / Roszko, Kelly L / Wang, Yan / Robey, Pamela G / Collins, Michael T / Boyce, Alison M / de Castro Diaz, Luis Fernandez

    bioRxiv : the preprint server for biology

    2024  

    Abstract: Fibrous dysplasia (FD) is a mosaic skeletal disorder caused by somatic activating variants ... ...

    Abstract Fibrous dysplasia (FD) is a mosaic skeletal disorder caused by somatic activating variants in
    Language English
    Publishing date 2024-02-28
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2024.02.23.581225
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  6. Article: Resistance to Naïve and Formative Pluripotency Conversion in RSeT Human Embryonic Stem Cells.

    Chen, Kevin G / Johnson, Kory R / Park, Kyeyoon / Maric, Dragan / Yang, Forest / Liu, Wen Fang / Fann, Yang C / Mallon, Barbara S / Robey, Pamela G

    bioRxiv : the preprint server for biology

    2024  

    Abstract: One of the most important properties of human embryonic stem cells (hESCs) is related to their primed and naïve pluripotent states. Our previous meta-analysis indicates the existence of heterogeneous pluripotent states derived from diverse naïve ... ...

    Abstract One of the most important properties of human embryonic stem cells (hESCs) is related to their primed and naïve pluripotent states. Our previous meta-analysis indicates the existence of heterogeneous pluripotent states derived from diverse naïve protocols. In this study, we have characterized a commercial medium (RSeT)-based pluripotent state under various growth conditions. Notably, RSeT hESCs can circumvent hypoxic growth conditions as required by naïve hESCs, in which some RSeT cells (e.g., H1 cells) exhibit much lower single cell plating efficiency, having altered or much retarded cell growth under both normoxia and hypoxia. Evidently, hPSCs lack many transcriptomic hallmarks of naïve and formative pluripotency (a phase between naive and primed states). Integrative transcriptome analysis suggests our primed and RSeT hESCs are close to the early stage of post-implantation embryos, similar to the previously reported primary hESCs and early hESC cultures. Moreover, RSeT hESCs did not express naïve surface markers such as CD75, SUSD2, and CD130 at a significant level. Biochemically, RSeT hESCs exhibit a differential dependency of FGF2 and co-independency of both Janus kinase (JAK) and TGFβ signaling in a cell-line-specific manner. Thus, RSeT hESCs represent a previously unrecognized pluripotent state downstream of formative pluripotency. Our data suggest that human naïve pluripotent potentials may be restricted in RSeT medium. Hence, this study provides new insights into pluripotent state transitions
    Language English
    Publishing date 2024-04-12
    Publishing country United States
    Document type Preprint
    DOI 10.1101/2024.02.16.580778
    Database MEDical Literature Analysis and Retrieval System OnLINE

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  7. Article ; Online: Transcriptomic Signature and Pro-Osteoclastic Secreted Factors of Abnormal Bone-Marrow Stromal Cells in Fibrous Dysplasia.

    Michel, Zachary / Raborn, Layne N / Spencer, Tiahna / Pan, Kristen S / Martin, Daniel / Roszko, Kelly L / Wang, Yan / Robey, Pamela G / Collins, Michael T / Boyce, Alison M / de Castro, Luis Fernandez

    Cells

    2024  Volume 13, Issue 9

    Abstract: Fibrous dysplasia (FD) is a mosaic skeletal disorder caused by somatic activating variants ... ...

    Abstract Fibrous dysplasia (FD) is a mosaic skeletal disorder caused by somatic activating variants of
    MeSH term(s) Humans ; Animals ; Mesenchymal Stem Cells/metabolism ; Transcriptome/genetics ; Mice ; Fibrous Dysplasia of Bone/genetics ; Fibrous Dysplasia of Bone/metabolism ; Fibrous Dysplasia of Bone/pathology ; Male ; Female ; Cytokines/metabolism ; GTP-Binding Protein alpha Subunits, Gs/metabolism ; GTP-Binding Protein alpha Subunits, Gs/genetics ; Adult ; Middle Aged
    Chemical Substances Cytokines ; GTP-Binding Protein alpha Subunits, Gs (EC 3.6.5.1)
    Language English
    Publishing date 2024-04-30
    Publishing country Switzerland
    Document type Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
    ZDB-ID 2661518-6
    ISSN 2073-4409 ; 2073-4409
    ISSN (online) 2073-4409
    ISSN 2073-4409
    DOI 10.3390/cells13090774
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  8. Article ; Online: Bone Marrow Stromal Cell Assays: In Vitro and In Vivo.

    Robey, Pamela G / Kuznetsov, Sergei A / Bianco, Paolo / Riminucci, Mara

    Methods in molecular biology (Clifton, N.J.)

    2020  Volume 2230, Page(s) 379–396

    Abstract: Populations of bone marrow stromal cells (BMSCs, also known as bone marrow-derived "mesenchymal stem cells") contain a subset of cells that are able to recapitulate the formation of a bone/marrow organ (skeletal stem cells, SSCs). It is now apparent that ...

    Abstract Populations of bone marrow stromal cells (BMSCs, also known as bone marrow-derived "mesenchymal stem cells") contain a subset of cells that are able to recapitulate the formation of a bone/marrow organ (skeletal stem cells, SSCs). It is now apparent that cells with similar but not identical properties can be isolated from other skeletal compartments (growth plate, periosteum). The biological properties of BMSCs, and these related stem/progenitor cells, are assessed by a variety of assays, both in vitro and in vivo. Application of these assays in an appropriate fashion provide a great deal of information on the role of BMSCs, and the subset of SSCs, in health and in disease.
    MeSH term(s) Animals ; Bone Marrow Cells/ultrastructure ; Bone and Bones/ultrastructure ; Cell Differentiation/genetics ; Colony-Forming Units Assay/methods ; Growth Plate/ultrastructure ; Humans ; Mesenchymal Stem Cells/ultrastructure
    Language English
    Publishing date 2020-11-16
    Publishing country United States
    Document type Journal Article ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
    ISSN 1940-6029
    ISSN (online) 1940-6029
    DOI 10.1007/978-1-0716-1028-2_23
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  9. Article ; Online: From Stem Cells to Bone-Forming Cells.

    Donsante, Samantha / Palmisano, Biagio / Serafini, Marta / Robey, Pamela G / Corsi, Alessandro / Riminucci, Mara

    International journal of molecular sciences

    2021  Volume 22, Issue 8

    Abstract: Bone formation starts near the end of the embryonic stage of development and continues throughout life during bone modeling and growth, remodeling, and when needed, regeneration. Bone-forming cells, traditionally termed osteoblasts, produce, assemble, ... ...

    Abstract Bone formation starts near the end of the embryonic stage of development and continues throughout life during bone modeling and growth, remodeling, and when needed, regeneration. Bone-forming cells, traditionally termed osteoblasts, produce, assemble, and control the mineralization of the type I collagen-enriched bone matrix while participating in the regulation of other cell processes, such as osteoclastogenesis, and metabolic activities, such as phosphate homeostasis. Osteoblasts are generated by different cohorts of skeletal stem cells that arise from different embryonic specifications, which operate in the pre-natal and/or adult skeleton under the control of multiple regulators. In this review, we briefly define the cellular identity and function of osteoblasts and discuss the main populations of osteoprogenitor cells identified to date. We also provide examples of long-known and recently recognized regulatory pathways and mechanisms involved in the specification of the osteogenic lineage, as assessed by studies on mice models and human genetic skeletal diseases.
    MeSH term(s) Animals ; Bone and Bones/cytology ; Bone and Bones/embryology ; Epigenesis, Genetic ; Humans ; Osteoblasts/cytology ; Osteogenesis/genetics ; Signal Transduction ; Stem Cells/cytology
    Language English
    Publishing date 2021-04-13
    Publishing country Switzerland
    Document type Journal Article ; Review
    ZDB-ID 2019364-6
    ISSN 1422-0067 ; 1422-0067 ; 1661-6596
    ISSN (online) 1422-0067
    ISSN 1422-0067 ; 1661-6596
    DOI 10.3390/ijms22083989
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  10. Article ; Online: A single day of TGF-β1 exposure activates chondrogenic and hypertrophic differentiation pathways in bone marrow-derived stromal cells.

    Futrega, Kathryn / Robey, Pamela G / Klein, Travis J / Crawford, Ross W / Doran, Michael R

    Communications biology

    2021  Volume 4, Issue 1, Page(s) 29

    Abstract: Virtually all bone marrow-derived stromal cell (BMSC) chondrogenic induction cultures include greater than 2 weeks exposure to transforming growth factor-β (TGF-β), but fail to generate cartilage-like tissue suitable for joint repair. Herein we used a ... ...

    Abstract Virtually all bone marrow-derived stromal cell (BMSC) chondrogenic induction cultures include greater than 2 weeks exposure to transforming growth factor-β (TGF-β), but fail to generate cartilage-like tissue suitable for joint repair. Herein we used a micro-pellet model (5 × 10
    MeSH term(s) Bone Marrow Cells/physiology ; Cartilage, Articular/cytology ; Chondrocytes/physiology ; Chondrogenesis ; Humans ; Hypertrophy ; Sequence Analysis, RNA ; Tissue Engineering/methods ; Transforming Growth Factor beta1/physiology
    Chemical Substances TGFB1 protein, human ; Transforming Growth Factor beta1
    Language English
    Publishing date 2021-01-04
    Publishing country England
    Document type Comparative Study ; Evaluation Study ; Journal Article ; Research Support, N.I.H., Intramural ; Research Support, Non-U.S. Gov't
    ISSN 2399-3642
    ISSN (online) 2399-3642
    DOI 10.1038/s42003-020-01520-0
    Database MEDical Literature Analysis and Retrieval System OnLINE

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