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Development of rolled scaffold for high-density adherent cell culture

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Abstract

Bioreactors for large-scale culture of mammalian cells are playing vital roles in biotechnology and bioengineering. Various bioreactors have been developed, but their capacity and efficiency are often limited by insufficient mass transfer rate and high shear stress. A rolled scaffold (RS) is a fully defined scaffold for high-density adherent culture of mammalian cells. The RS is a polymer film with spacers, that is rolled into a cylinder with a pre-determined gap between each turn. Cells are cultured on its inner surfaces, while media flows through the gap. The RS exhibits high surface-area-to-volume ratio over 100 cm2/mL and can transport nutrients and gases with significantly reduced shear stress via convection in a unidirectional laminar flow, rather than diffusion and random turbulent flow as in stirred-tank bioreactors. In this paper, we expanded Chinese Hamster Ovary cells with RS bioreactors and demonstrated cell culture density over 60 million cells/mL with a growth rate higher than conventional suspension culture. Besides, murine embryonic stem cells were successfully expanded without losing their pluripotency. The RS will provide an affordable, scalable, and reliable platform for large-scale culture of recombinant cells in biopharmaceutical industries and shear-sensitive stem cells for tissue engineering.

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Acknowledgements

The ES cell line used for this research project, E14 RP11-88 L12 NKX2-5-EmGFP ES cell line, RRID:MMRRC_030473-UCD, was obtained from the Mutant Mouse Resource & Research Centers, a NCRR-NIH funded strain repository, and was donated to the MMRRC by Dr. Bruce Conklin, M.D., Gladstone Institute of Cardiovascular Disease. A. Mehrnezhad was supported from the Louisiana Board of Regents (Grant LEQSF(2014-17)-RD-A-05) and Katie M. Hamel was supported by the Flagship Graduate Fellowship.

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Authors and Affiliations

  1. Division of Electrical and Computer Engineering, Louisiana State University, Baton Rouge, LA, 70803, USA

    Ashkan YekrangSafakar, Ali Mehrnezhad & Kidong Park

  2. Department of Biological Engineering, Louisiana State University, Baton Rouge, LA, USA

    Katie M. Hamel & Jangwook P. Jung

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  1. Ashkan YekrangSafakar
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  2. Katie M. Hamel
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  3. Ali Mehrnezhad
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  4. Jangwook P. Jung
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  5. Kidong Park
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Contributions

K.P. conceived RS and RS-bioreactor; K.P., A.Y., and A.M. built experimental setups; A.Y. collected data on RS culture; K.P., A.Y., and J.P.J. designed experiments; J.P.J. and K.M.H. provided and characterized mESCs.

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Correspondence to Kidong Park.

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A patent application has been filed on this technology by Louisiana State University.

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YekrangSafakar, A., Hamel, K.M., Mehrnezhad, A. et al. Development of rolled scaffold for high-density adherent cell culture. Biomed Microdevices 22, 4 (2020). https://doi.org/10.1007/s10544-019-0459-9

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