Menaquinone-7 Supplementation Improves Osteogenesis in Pluripotent Stem Cell Derived Mesenchymal Stem Cells

A.C. Akbulut; G.B. Wasilewski; N. Rapp; F. Forin; H. Singer; K.J. Czogalla-Nitsche; L.J. Schurgers

doi:10.3389/fcell.2020.618760

Menaquinone-7 Supplementation Improves Osteogenesis in Pluripotent Stem Cell Derived Mesenchymal Stem Cells

A.C. Akbulut, G.B. Wasilewski, N. Rapp, F. Forin, H. Singer, K.J. Czogalla-Nitsche, L.J. Schurgers^*

^*Corresponding author for this work

Research output: Contribution to journal › Article › Academic › peer-review

Abstract

Development of clinical stem cell interventions are hampered by immature cell progeny under current protocols. Human mesenchymal stem cells (hMSCs) are characterized by their ability to self-renew and differentiate into multiple lineages. Generating hMSCs from pluripotent stem cells (iPSCs) is an attractive avenue for cost-efficient and scalable production of cellular material. In this study we generate mature osteoblasts from iPSCs using a stable expandable MSC intermediate, refining established protocols. We investigated the timeframe and phenotype of cells under osteogenic conditions as well as the effect of menaquinone-7 (MK-7) on differentiation. From day 2 we noted a significant increase in RUNX2 expression under osteogenic conditions with MK-7, as well as decreases in ROS species production, increased cellular migration and changes to dynamics of collagen deposition when compared to differentiated cells that were not treated with MK-7. At day 21 OsteoMK-7 increased alkaline phosphatase activity and collagen deposition, as well as downregulated RUNX2 expression, suggesting to a mature cellular phenotype. Throughout we note no changes to expression of osteocalcin suggesting a non-canonical function of MK-7 in osteoblast differentiation. Together our data provide further mechanistic insight between basic and clinical studies on extrahepatic activity of MK-7. Our findings show that MK-7 promotes osteoblast maturation thereby increasing osteogenic differentiation.

Original language	English
Article number	618760
Number of pages	13
Journal	Frontiers in Cell and Developmental Biology
Volume	8
DOIs	https://doi.org/10.3389/fcell.2020.618760
Publication status	Published - 28 Jan 2021

Keywords

menaquinone-7
mesenchymal stem cells
osteogenesis
pluripotent stem cells
vitamin k
vitamin K

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10.3389/fcell.2020.618760Licence: CC BY

Cite this

@article{cbecacd7752d4a5eab1734a1b4b1ac42,

title = "Menaquinone-7 Supplementation Improves Osteogenesis in Pluripotent Stem Cell Derived Mesenchymal Stem Cells",

abstract = "Development of clinical stem cell interventions are hampered by immature cell progeny under current protocols. Human mesenchymal stem cells (hMSCs) are characterized by their ability to self-renew and differentiate into multiple lineages. Generating hMSCs from pluripotent stem cells (iPSCs) is an attractive avenue for cost-efficient and scalable production of cellular material. In this study we generate mature osteoblasts from iPSCs using a stable expandable MSC intermediate, refining established protocols. We investigated the timeframe and phenotype of cells under osteogenic conditions as well as the effect of menaquinone-7 (MK-7) on differentiation. From day 2 we noted a significant increase in RUNX2 expression under osteogenic conditions with MK-7, as well as decreases in ROS species production, increased cellular migration and changes to dynamics of collagen deposition when compared to differentiated cells that were not treated with MK-7. At day 21 OsteoMK-7 increased alkaline phosphatase activity and collagen deposition, as well as downregulated RUNX2 expression, suggesting to a mature cellular phenotype. Throughout we note no changes to expression of osteocalcin suggesting a non-canonical function of MK-7 in osteoblast differentiation. Together our data provide further mechanistic insight between basic and clinical studies on extrahepatic activity of MK-7. Our findings show that MK-7 promotes osteoblast maturation thereby increasing osteogenic differentiation.",

keywords = "menaquinone-7, mesenchymal stem cells, osteogenesis, pluripotent stem cells, vitamin k, vitamin K",

author = "A.C. Akbulut and G.B. Wasilewski and N. Rapp and F. Forin and H. Singer and K.J. Czogalla-Nitsche and L.J. Schurgers",

note = "Funding Information: We would like to thank Dr. de Kok from Department of Toxicogenomics, Maastricht University for providing us with primers for P21. Funding. Research was in part funded via the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 722609 (AA and LS) and grant agreement No 764474 (NR and LS). Research from GW and LS was in part funded by the Norwegian Research Council (Project 241584) and NattoPharma ASA. This work was supported in part by the Emmy-Noether grant from the Deutsche Forschungsgemeinschaft (Project 405008087). Funding Information: Research was in part funded via the European Union{\textquoteright}s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 722609 (AA and LS) and grant agreement No 764474 (NR and LS). Research from GW and LS was in part funded by the Norwegian Research Council (Project 241584) and NattoPharma ASA. This work was supported in part by the Emmy-Noether grant from the Deutsche Forschungsgemeinschaft (Project 405008087). Publisher Copyright: {\textcopyright} Copyright {\textcopyright} 2021 Akbulut, Wasilewski, Rapp, Forin, Singer, Czogalla-Nitsche and Schurgers.",

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doi = "10.3389/fcell.2020.618760",

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T1 - Menaquinone-7 Supplementation Improves Osteogenesis in Pluripotent Stem Cell Derived Mesenchymal Stem Cells

AU - Akbulut, A.C.

AU - Wasilewski, G.B.

AU - Rapp, N.

AU - Forin, F.

AU - Singer, H.

AU - Czogalla-Nitsche, K.J.

AU - Schurgers, L.J.

N1 - Funding Information: We would like to thank Dr. de Kok from Department of Toxicogenomics, Maastricht University for providing us with primers for P21. Funding. Research was in part funded via the European Union's Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 722609 (AA and LS) and grant agreement No 764474 (NR and LS). Research from GW and LS was in part funded by the Norwegian Research Council (Project 241584) and NattoPharma ASA. This work was supported in part by the Emmy-Noether grant from the Deutsche Forschungsgemeinschaft (Project 405008087). Funding Information: Research was in part funded via the European Union’s Horizon 2020 research and innovation programme under the Marie Sklodowska-Curie grant agreement No 722609 (AA and LS) and grant agreement No 764474 (NR and LS). Research from GW and LS was in part funded by the Norwegian Research Council (Project 241584) and NattoPharma ASA. This work was supported in part by the Emmy-Noether grant from the Deutsche Forschungsgemeinschaft (Project 405008087). Publisher Copyright: © Copyright © 2021 Akbulut, Wasilewski, Rapp, Forin, Singer, Czogalla-Nitsche and Schurgers.

PY - 2021/1/28

Y1 - 2021/1/28

N2 - Development of clinical stem cell interventions are hampered by immature cell progeny under current protocols. Human mesenchymal stem cells (hMSCs) are characterized by their ability to self-renew and differentiate into multiple lineages. Generating hMSCs from pluripotent stem cells (iPSCs) is an attractive avenue for cost-efficient and scalable production of cellular material. In this study we generate mature osteoblasts from iPSCs using a stable expandable MSC intermediate, refining established protocols. We investigated the timeframe and phenotype of cells under osteogenic conditions as well as the effect of menaquinone-7 (MK-7) on differentiation. From day 2 we noted a significant increase in RUNX2 expression under osteogenic conditions with MK-7, as well as decreases in ROS species production, increased cellular migration and changes to dynamics of collagen deposition when compared to differentiated cells that were not treated with MK-7. At day 21 OsteoMK-7 increased alkaline phosphatase activity and collagen deposition, as well as downregulated RUNX2 expression, suggesting to a mature cellular phenotype. Throughout we note no changes to expression of osteocalcin suggesting a non-canonical function of MK-7 in osteoblast differentiation. Together our data provide further mechanistic insight between basic and clinical studies on extrahepatic activity of MK-7. Our findings show that MK-7 promotes osteoblast maturation thereby increasing osteogenic differentiation.

AB - Development of clinical stem cell interventions are hampered by immature cell progeny under current protocols. Human mesenchymal stem cells (hMSCs) are characterized by their ability to self-renew and differentiate into multiple lineages. Generating hMSCs from pluripotent stem cells (iPSCs) is an attractive avenue for cost-efficient and scalable production of cellular material. In this study we generate mature osteoblasts from iPSCs using a stable expandable MSC intermediate, refining established protocols. We investigated the timeframe and phenotype of cells under osteogenic conditions as well as the effect of menaquinone-7 (MK-7) on differentiation. From day 2 we noted a significant increase in RUNX2 expression under osteogenic conditions with MK-7, as well as decreases in ROS species production, increased cellular migration and changes to dynamics of collagen deposition when compared to differentiated cells that were not treated with MK-7. At day 21 OsteoMK-7 increased alkaline phosphatase activity and collagen deposition, as well as downregulated RUNX2 expression, suggesting to a mature cellular phenotype. Throughout we note no changes to expression of osteocalcin suggesting a non-canonical function of MK-7 in osteoblast differentiation. Together our data provide further mechanistic insight between basic and clinical studies on extrahepatic activity of MK-7. Our findings show that MK-7 promotes osteoblast maturation thereby increasing osteogenic differentiation.

KW - menaquinone-7

KW - mesenchymal stem cells

KW - osteogenesis

KW - pluripotent stem cells

KW - vitamin k

KW - vitamin K

U2 - 10.3389/fcell.2020.618760

DO - 10.3389/fcell.2020.618760

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SN - 2296-634X

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JO - Frontiers in Cell and Developmental Biology

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