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Fluid Flow Modulation of Murine Embryonic Stem Cell Pluripotency Gene Expression in the Absence of LIF

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Abstract

Fluid forces are strong modulators of cell fate and fundamental components of spinner flask bioreactors used for stem cell expansion and differentiation. Here, we investigated the effects of fluid forces on murine embryonic stem cells (mESCs) in the absence of Leukemia Inhibitory Factor (LIF) using parallel-plate flow chambers. Cells were seeded onto gelatin-coated glass slides and grown for 2.5 days before exposure to fluid forces. Pluripotency marker gene expression was quantified by qPCR. An average shear stress of 0.6 Pa applied for 24 h in the absence of LIF and presence of high molecular weight dextran increased Oct4 and Sox2, decreased Nanog, and did not change in Rex1 mRNA levels in comparison to statically cultured cells in the presence of LIF. At 0.3 Pa shear stress, Oct4 and Sox2 expression increased, with a reduction in Nanog and Rex1 levels. The presence of pulsation significantly increased expression of Rex1 and Nanog, but not expression of Oct4 or Sox2, compared to cells exposed to steady flow for 24 h. This study suggests incorporation of high cell–cell contact, viscosity elevation with dextran, moderate shear stress (0.6 Pa), and the presence of pulsatility in bioreactor expansion protocols for mESCs to support maintenance of pluripotency.

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Acknowledgments

Funding for this study was provided by the Canadian Institutes of Health Research (CIHR), the Natural Sciences and Engineering Research Council of Canada, and the Department of Chemical and Petroleum Engineering at the University of Calgary. Equipment funding was provided by the Canada Foundation for Innovation and the Alberta Advanced Education and Technology Science and Research Investments Program. GGL and OH held scholarships from the Skeletal Regenerative Medicine Team, funded by CIHR.

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Authors and Affiliations

  1. Cellular and Molecular Bioengineering Research Laboratory, Schulich School of Engineering, University of Calgary, Calgary, AB, T2N 1N4, Canada

    Giovanna G. Lara, Olesja Hazenbiller, Robert D. Shepherd & Kristina D. Rinker

  2. Biomedical Engineering Graduate Program, University of Calgary, Calgary, AB, T2N 1N4, Canada

    Giovanna G. Lara, Michael S. Kallos & Kristina D. Rinker

  3. Pharmaceutical Production Research Facility, University of Calgary, Calgary, AB, T2N 1N4, Canada

    Tia Gareau & Michael S. Kallos

  4. Centre for Bioengineering Research and Education, Schulich School of Engineering, University of Calgary, 2500 University Dr. N.W., Calgary, AB, T2N 1N4, Canada

    Robert D. Shepherd, Michael S. Kallos & Kristina D. Rinker

  5. Department of Chemical and Petroleum Engineering, Schulich School of Engineering, University of Calgary, Calgary, AB, T2N 1N4, Canada

    Robert D. Shepherd, Michael S. Kallos & Kristina D. Rinker

  6. Department of Biochemistry and Molecular Biology, Faculty of Medicine; University of Calgary, Calgary, AB, T2N 4Z6, Canada

    Derrick E. Rancourt

  7. Department of Physiology and Pharmacology, Faculty of Medicine; University of Calgary, Calgary, AB, T2N 4N1, Canada

    Kristina D. Rinker

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  1. Giovanna G. Lara
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Correspondence to Kristina D. Rinker.

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Associate Editor Chwee Teck Lim oversaw the review of this article.

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Lara, G.G., Hazenbiller, O., Gareau, T. et al. Fluid Flow Modulation of Murine Embryonic Stem Cell Pluripotency Gene Expression in the Absence of LIF. Cel. Mol. Bioeng. 6, 335–345 (2013). https://doi.org/10.1007/s12195-013-0287-6

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