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Stiffness-Controlled Hydrogels for 3D Cell Culture Models.

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Summary

Freeze-dried nanofibrillated cellulose (NFC) hydrogel enables easy one-step 3D cell spheroid culturing. This biomaterial supports cell viability and morphology, paving the way for automated cell culture analysis.

Keywords:
3D cell culturebiomaterialsconvolutional neural networkfreeze-dryinghydrogelnanofibrillated cellulose

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Area of Science:

  • Biomaterials Science
  • Cell Biology
  • Biotechnology

Background:

  • Nanofibrillated cellulose (NFC) hydrogel is a versatile biomaterial.
  • Freeze-drying NFC hydrogel creates highly porous structures.
  • These porous structures can be reconstituted to tune mechanical properties.

Purpose of the Study:

  • To investigate the use of freeze-dried and reconstituted NFC hydrogel for 3D cell spheroid culturing.
  • To evaluate cell viability, morphology, and metabolic activity in 3D spheroids cultured in NFC hydrogel.
  • To explore the application of automated cell segmentation in 3D cultured spheroids.

Main Methods:

  • NFC hydrogel was freeze-dried and reconstituted into various concentrations.
  • Primary mesenchymal stem/stromal cells, PC3, and HepG2 cells were cultured as 3D spheroids.
  • Cell viability, morphology, and metabolic activity were assessed.
  • A convolutional neural network (CNN) was used for automatic nuclei segmentation.

Main Results:

  • Freeze-dried and reconstituted NFC hydrogel successfully supported one-step 3D cell spheroid culturing.
  • No significant differences in cell viability or morphology were observed compared to fresh NFC hydrogel.
  • 3D cultured spheroids exhibited stable metabolic activity and high viability (>99%).
  • CNN-based segmentation effectively analyzed individual cells within 3D spheroids.

Conclusions:

  • Freeze-dried NFC hydrogel offers a straightforward method for 3D cell spheroid culture.
  • The material is suitable for culturing various cell types, maintaining cell health.
  • This approach facilitates the automation of 3D cell culturing and subsequent analysis.