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The Role of Primary Cilia in Mesenchymal Stem Cell Differentiation: A Pivotal Switch in Guiding Lineage Commitment

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Abstract

Primary cilia are sensory organelles that have been shown to play a critical role in lineage commitment. It was our hypothesis that the primary cilium is necessary for chemically induced differentiation of human mesenchymal stem cells (MSC). To investigate this, polaris siRNA was used to inhibit the primary cilia and the mRNA levels of transcription factors Runx2, PPARγ were measured by RT PCR as markers of osteogenic, adipogenic and chondrogenic differentiation, respectively. MSCs with inhibited primary cilia had significantly decreased basal mRNA expression levels of all three lineages specific transcription factors indicating that primary cilia are critical in multiple differentiation pathways. Furthermore, to determine if primary cilia play a role in the differentiation potential of MSCs, progenitor cells transfected with either scrambled or polaris siRNA were cultured in osteo-inductive, chondro-inductive, or adipo-inductive media and lineage commitment was ascertained. Interestingly, within 24 h of culture, cells transfected with polaris siRNA in both osteogenic and adipogenic media lost adhesion and released from the slides; however MSCs in chondrogenic media as well as cells transfected with scrambled siRNA did not. These results suggest that the primary cilium is necessary for the normal progression of chemically induced osteogenic and adipogenic differentiation. As a control, the experiment was repeated with NIH3T3 fibroblasts and none of the effects of inhibited primary cilia were observed indicating that the loss of adhesion may be specific to MSCs. Furthermore after biochemically inducing the cells to differentiate, polaris knockdown resulted in abrogation of both Runx2 and PPARγ mRNA while SOX9 mRNA expression was significantly lower. These results suggest that primary cilia play an essential role not only in the initiation of both osteogenic and adipogenic differentiation, but also in maintaining the phenotype of differentiated cells. Interestingly, chondrogenic differentiation appeared less dependent on a functional primary cilium.

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Acknowledgments

This work was funded by National Institute of Health Grants AR45989 and AR54156 and the United States Department of Veterans Affairs.

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Authors and Affiliations

  1. Department of Veterans Affairs, Bone and Joint Rehabilitation Center, Palo Alto, CA, USA

    Padmaja Tummala, Emily J. Arnsdorf & Christopher R. Jacobs

  2. Department of Bioengineering, Stanford University, Stanford, CA, USA

    Emily J. Arnsdorf & Christopher R. Jacobs

  3. InCube Labs, Menlo Park, CA, USA

    Emily J. Arnsdorf

  4. MedImmune Vaccines, Mountain View, CA, USA

    Padmaja Tummala

  5. Department of Mechanical Engineering, Stanford University, Stanford, CA, USA

    Christopher R. Jacobs

  6. Department of Biomedical Engineering, Columbia University, 500 W. 120th Street, 351 Engineering Terrace, New York, NY, 10027, USA

    Christopher R. Jacobs

Authors
  1. Padmaja Tummala
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  2. Emily J. Arnsdorf
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  3. Christopher R. Jacobs
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Correspondence to Christopher R. Jacobs.

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Associate Editor Lance Kam and Partha Roy oversaw the review of this article.

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Tummala, P., Arnsdorf, E.J. & Jacobs, C.R. The Role of Primary Cilia in Mesenchymal Stem Cell Differentiation: A Pivotal Switch in Guiding Lineage Commitment. Cel. Mol. Bioeng. 3, 207–212 (2010). https://doi.org/10.1007/s12195-010-0127-x

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  • DOI: https://doi.org/10.1007/s12195-010-0127-x

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