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Non-Enzymatic Cell Expansion and Harvesting Using a Smart Thermo-Responsive Gel.

Zhiyu Yan1, Nuno Honrado1, Naiwen Tan1

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Summary

This study introduces temperature-responsive alginate beads with poly(N-isopropylacrylamide) (PNIPAAm) as an enzyme-free method for harvesting mammalian cells. This approach preserves cell viability and extracellular matrix, offering a promising alternative to traditional trypsinization for cell manufacturing.

Keywords:
PNIPAAmalginate hydrogel beadscell therapynon-enzymatic cell harvestingthermo-responsive polymer

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Area of Science:

  • Biotechnology and Regenerative Medicine
  • Materials Science
  • Cell Biology

Background:

  • Advanced cell-based therapies require large numbers of viable mammalian cells.
  • Conventional enzymatic harvesting (e.g., trypsinization) can damage cells and reduce viability.
  • There is a need for gentler, non-enzymatic cell harvesting methods.

Purpose of the Study:

  • To develop and evaluate a temperature-responsive alginate bead system for enzyme-free mammalian cell harvesting.
  • To assess the impact of this method on cell integrity, viability, and extracellular matrix preservation.
  • To explore its potential for advanced cell manufacturing and therapeutic applications.

Main Methods:

  • Synthesis of alginate hydrogel beads incorporating poly(N-isopropylacrylamide) (PNIPAAm).
  • Characterization of beads using SEM, FTIR, DSC, and µ-CT.
  • Culture of mouse fibroblast cells (L929) on beads and assessment of proliferation and viability (CCK-8, Live/Dead assays).

Main Results:

  • PNIPAAm-modified alginate beads demonstrated temperature-responsive properties.
  • Significant cell proliferation and high viability were observed over seven days of culture.
  • The method successfully enabled temperature-controlled cell detachment, preserving cellular structure.

Conclusions:

  • PNIPAAm-modified alginate beads offer a promising enzyme-free platform for mammalian cell harvesting.
  • This technology has the potential to improve cell viability and preserve extracellular matrix compared to trypsin.
  • The system is suitable for applications in advanced cell manufacturing and therapeutic delivery.