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Methylcellulose Based Thermally Reversible Hydrogels

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Adipose-Derived Stem Cells

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1773))

Abstract

The creation of single and multilayered adult stem cells (ASCs) sheets is presented. The stem cell sheets preserve the cell–cell and cell–extracellular matrices and are developed by utilizing a thermally reversible methylcellulose (MC) coated tissue culture polystyrene (TCPS) dish. This technique is an improvement and a simplification of earlier noninvasive cell retrieval methods based on the use of a temperature-responsive poly(N-isopropylacrylamide) (PIPAAm) coated TCPS dishes. The optimal combination of MC–water–salt was determined to be 12–14% of MC (mol. wt. of 15,000) in water with 0.5× PBS (~150 mOsm). This solution exhibited a gel formation temperature of ~32 °C. The addition (evenly spread) of 1 ml of 3 mg/ml rat tail type-I (pH adjusted to 7.5) over the MC coated surface at 37 °C improves ASC adhesion and proliferation on the methylcellulose system. Upon confluence, a continuous monolayer ASC sheet was formed on the surface of the MC hydrogel system. When the grown cell sheet was removed from the incubator and exposed to room temperature (~30 °C), it spontaneously and gradually detached from the surface of the thermoresponsive hydrogel, creating an ASC sheet.

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Acknowledgments

The authors thank Dr. Elizabeth Clubb and Dr. James Wade at the Pennington Biomedical Research Center (PBRC) for supplying the liposuction aspirates and their many patients for consenting to participate in this protocol. Acknowledgements are also due to Dr. Gimble, a longtime colleague and friend without whose help this work would never have been initiated. And finally, acknowledgements are also due to Dr. S. Thirumala for performing the seminal experiments reported in this chapter. This work was supported in part by funding from the Louisiana Board of Regents and the Department of Mechanical Engineering at the Louisiana State University (LSU). Anoosha Forghani was supported the NSF EPSCoR LA-SiGMA project under award #EPS-1003897.

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Authors and Affiliations

  1. Bioengineering Laboratory, Department of Mechanical Engineering, Louisiana State University, Baton Rouge, LA, USA

    Anoosha Forghani & Ram Devireddy

Authors
  1. Anoosha Forghani
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  2. Ram Devireddy
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Corresponding author

Correspondence to Ram Devireddy .

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Editors and Affiliations

  1. Center for Stem Cell Research and Regenerative Medicine, Tulane University School of Medicine, New Orleans, LA, USA

    Bruce A. Bunnell  & Jeffrey M. Gimble  & 

  2. LaCell LLC, New Orleans, LA, USA

    Jeffrey M. Gimble

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Forghani, A., Devireddy, R. (2018). Methylcellulose Based Thermally Reversible Hydrogels. In: Bunnell, B.A., Gimble, J.M. (eds) Adipose-Derived Stem Cells. Methods in Molecular Biology, vol 1773. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-7799-4_5

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  • DOI: https://doi.org/10.1007/978-1-4939-7799-4_5

  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-7797-0

  • Online ISBN: 978-1-4939-7799-4

  • eBook Packages: Springer Protocols

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