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Thermo-responsive methylcellulose hydrogels as temporary substrate for cell sheet biofabrication

  • Tissue Engineering Constructs and Cell Substrates
  • Original Research
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Abstract

Methylcellulose (MC), a water-soluble polymer derived from cellulose, was investigated as a possible temporary substrate having thermo-responsive properties favorable for cell culturing. MC-based hydrogels were prepared by a dispersion technique, mixing MC powder (2, 4, 6, 8, 10, 12 % w/v) with selected salts (sodium sulphate, Na2SO4), sodium phosphate, calcium chloride, or phosphate buffered saline, to evaluate the influence of different compositions on the thermo-responsive behavior. The inversion test was used to determine the gelation temperatures of the different hydrogel compositions; thermo-mechanical properties and thermo-reversibility of the MC hydrogels were investigated by rheological analysis. Gelation temperatures and rheological behavior depended on the MC concentration and type and concentration of salt used in hydrogel preparation. In vitro cytotoxicity tests, performed using L929 mouse fibroblasts, showed no toxic release from all the tested hydrogels. Among the investigated compositions, the hydrogel composed of 8 % w/v MC with 0.05 M Na2SO4 had a thermo-reversibility temperature at 37 °C. For that reason, this formulation was thus considered to verify the possibility of inducing in vitro spontaneous detachment of cells previously seeded on the hydrogel surface. A continuous cell layer (cell sheet) was allowed to grow and then detached from the hydrogel surface without the use of enzymes, thanks to the thermo-responsive behavior of the MC hydrogel. Immunofluorescence observation confirmed that the detached cell sheet was composed of closely interacting cells.

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Acknowledgments

The authors would like to thank Prof. Antonia Follenzi for kindly providing transduced NIH GFP cells and Dr. Frances Cooper for language revision.

Funding

A. Cochis was partially supported by PRIN 2010e2011 (PRIN 20102ZLNJ5_006), financed by the Ministry of Education, University and Research (M.I.U.R.), Rome, Italy.

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Authors and Affiliations

  1. Dipartimento di Chimica, Materiali e Ingegneria Chimica “G. Natta”, Politecnico di Milano, Piazza L. Da Vinci 32, Milan, MI, Italy

    Lina Altomare, Andrea Carletta & Silvia Farè

  2. INSTM, Consorzio Nazionale di Scienza e Tecnologia dei Materiali, Local Unit Politecnico di Milano, Milan, Italy

    Lina Altomare & Silvia Farè

  3. Dipartimento di Scienze della Salute, Università del Piemonte Orientale (UPO), Via Solaroli 17, 28100, Novara, NO, Italy

    Andrea Cochis & Lia Rimondini

  4. INSTM, Consorzio Nazionale di Scienza e Tecnologia dei Materiali, Local Unit Università del Piemonte Orientale, Novara, Italy

    Andrea Cochis & Lia Rimondini

Authors
  1. Lina Altomare
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  2. Andrea Cochis
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  3. Andrea Carletta
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  4. Lia Rimondini
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  5. Silvia Farè
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Correspondence to Lia Rimondini.

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Altomare, L., Cochis, A., Carletta, A. et al. Thermo-responsive methylcellulose hydrogels as temporary substrate for cell sheet biofabrication. J Mater Sci: Mater Med 27, 95 (2016). https://doi.org/10.1007/s10856-016-5703-8

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  • DOI: https://doi.org/10.1007/s10856-016-5703-8

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