Use of RUNX2 Expression to Identify Osteogenic Progenitor Cells Derived from Human Embryonic Stem Cells

Zou, Li; Kidwai, Fahad; Kopher, Ross A.; Motl, Jason A.; Kellum, Cory A.; Kaufman, Dan S.

doi:10.1016/j.stemcr.2015.01.008

Cited by 38 publications

(58 citation statements)

References 29 publications

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“…The expression of surface markers (CD73, CD90, and CD105), data from gene expression analysis, and Alizarin Red and immunofluorescent staining demonstrate the osteogenic commitment and differentiation potential of these GFP-expressing hESCs. These results are in agreement with previous studies [10, 15, 20, 34, 35]. These data also suggest the stable expression of GFP in hESC-OPs and the genetic manipulation did not affect their osteogenic differentiation.…”

Section: Discussionsupporting

confidence: 93%

“…These cell lines effectively report for pluripotency and hence play a role in detection of PSCs from their differentiated progenies. Similarly, Zou et al generated a RUNX2 promoter driven YFP based reporter system in hESCs to track their commitment to osteogenic lineage [15]. Using these transgenic hESCs, they elegantly demonstrated that BMP2 does not induce osteogenic differentiation but promotes the generation of CD73+ osteoprogenitors and their osteogenic differentiation.…”

Section: Discussionmentioning

confidence: 99%

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In Vitro Osteogenic Potential of Green Fluorescent Protein Labelled Human Embryonic Stem Cell‐Derived Osteoprogenitors

Islam

Sriram

et al. 2016

Stem Cells International

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Cellular therapy using stem cells in bone regeneration has gained increasing interest. Various studies suggest the clinical utility of osteoprogenitors-like mesenchymal stem cells in bone regeneration. However, limited availability of mesenchymal stem cells and conflicting evidence on their therapeutic efficacy limit their clinical application. Human embryonic stem cells (hESCs) are potentially an unlimited source of healthy and functional osteoprogenitors (OPs) that could be utilized for bone regenerative applications. However, limited ability to track hESC-derived progenies in vivo greatly hinders translational studies. Hence, in this study, we aimed to establish hESC-derived OPs (hESC-OPs) expressing green fluorescent protein (GFP) and to investigate their osteogenic differentiation potential in vitro. We fluorescently labelled H9-hESCs using a plasmid vector encoding GFP. The GFP-expressing hESCs were differentiated into hESC-OPs. The hESC-OPsGFP+ stably expressed high levels of GFP, CD73, CD90, and CD105. They possessed osteogenic differentiation potential in vitro as demonstrated by increased expression of COL1A1, RUNX2, OSTERIX, and OPG transcripts and mineralized nodules positive for Alizarin Red and immunocytochemical expression of osteocalcin, alkaline phosphatase, and collagen-I. In conclusion, we have demonstrated that fluorescently labelled hESC-OPs can maintain their GFP expression for the long term and their potential for osteogenic differentiation in vitro. In future, these fluorescently labelled hESC-OPs could be used for noninvasive assessment of bone regeneration, safety, and therapeutic efficacy.

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Section: Discussionsupporting

confidence: 93%

Section: Discussionmentioning

confidence: 99%

In Vitro Osteogenic Potential of Green Fluorescent Protein Labelled Human Embryonic Stem Cell‐Derived Osteoprogenitors

Islam

Sriram

et al. 2016

Stem Cells International

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“…This possibility is further corroborated by a recent manuscript in which RUNX2 was identified as one of the miR‐1305 targets . RUNX2 is an important osteogenic regulator shown to regulate the length of G1 phase of the cell cycle and apoptosis in highly proliferating mammary epithelial cells . Given that miR‐1305 is also involved in cell cycle regulation and apoptosis in pluripotent stem cells, it would be of interest to further investigate whether these two processes are mediated via regulation of RUNX2 .…”

Section: Discussionmentioning

confidence: 60%

A Novel Role for miR-1305 in Regulation of Pluripotency-Differentiation Balance, Cell Cycle, and Apoptosis in Human Pluripotent Stem Cells

et al. 2016

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Human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs) are defined as pluripotent in view of their self‐renewal ability and potential to differentiate to cells of all three germ layers. Recent studies have indicated that microRNAs (miRNAs) play an important role in the maintenance of pluripotency and cell cycle regulation. We used a microarray based approach to identify miRNAs that were enriched in hESCs when compared to differentiated cells and at the same time showed significant expression changes between different phases of cell cycle. We identified 34 candidate miRNAs and performed functional studies on one of these, miR‐1305, which showed the highest expression change during cell cycle transition. Overexpression of miR‐1305 induced differentiation of pluripotent stem cells, increased cell apoptosis and sped up G1/S transition, while its downregulation facilitated the maintenance of pluripotency and increased cell survival. Using target prediction software and luciferase based reporter assays we identified POLR3G as a downstream target by which miR‐1305 regulates the fine balance between maintenance of pluripotency and onset of differentiation. Overexpression of POLR3G rescued pluripotent stem cell differentiation induced by miR‐1305 overexpression. In contrast, knock‐down of POLR3G expression abolished the miR‐1305‐knockdown mediated enhancement of pluripotency, thus validating its role as miR‐1305 target in human pluripotent stem cells. Together our data point to an important role for miR‐1305 as a novel regulator of pluripotency, cell survival and cell cycle and uncovers new mechanisms and networks by which these processes are intertwined in human pluripotent stem cells. Stem Cells 2016;34:2306–2317

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“…This suggests that there is an uncertain threshold above which Mg induces differentiation, either by itself or in the presence of Sr (and minor reduction in Ca). RUNX2 expression typically increases during osteogenic differentiation, 33,34 but can also remain constant. 35 Extract 1 has a significantly increased RUNX2 expression [ Figure 3(B)], confirming signs of differentiation thus complementing the ALPL expression results [ Figure 3(A)].…”

Section: Discussionmentioning

confidence: 99%

The effect of Mg–Ca–Sr alloy degradation products on human mesenchymal stem cells

et al. 2017

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Biodegradable Mg alloys have the potential to replace currently used metallic medical implant devices, likely eliminating toxicity concerns and the need for secondary surgeries, while also providing a potentially stimulating environment for tissue growth. A recently developed Mg–Ca–Sr alloy possesses advantageous characteristics over other Mg alloys, having a good combination of strength and degradation behavior, while also displaying potentially osteogenic properties. To better understand the effect of alloy degradation products on cellular mechanisms, in vitro studies using human bone marrow‐derived mesenchymal stem cells were conducted. Ionic products of alloy dissolution were found to be nontoxic but changed the proliferation profile of stem cells. Furthermore, their presence changed the progress of osteogenic development, while concentrations of Mg in particular appeared to induce stem cell differentiation. The work presented herein provides a foundation for future alloy design where structures can be tailored to obtain specific implant performance. These potentially bioactive implants would reduce the risks for patients by shortening their healing time, minimizing discomfort and toxicity concerns, while reducing hospital costs. © 2017 The Authors Journal of Biomedical Materials Research Part B: Applied Biomaterials Published by Wiley Periodicals, Inc. J Biomed Mater Res Part B: Appl Biomater, 106B: 697–704, 2018.

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Use of RUNX2 Expression to Identify Osteogenic Progenitor Cells Derived from Human Embryonic Stem Cells

Cited by 38 publications

References 29 publications

In Vitro Osteogenic Potential of Green Fluorescent Protein Labelled Human Embryonic Stem Cell‐Derived Osteoprogenitors

In Vitro Osteogenic Potential of Green Fluorescent Protein Labelled Human Embryonic Stem Cell‐Derived Osteoprogenitors

A Novel Role for miR-1305 in Regulation of Pluripotency-Differentiation Balance, Cell Cycle, and Apoptosis in Human Pluripotent Stem Cells

The effect of Mg–Ca–Sr alloy degradation products on human mesenchymal stem cells

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