Stiffness-Controlled Hydrogels for 3D Cell Culture Models

Merivaara A, Koivunotko E, Manninen K, Kaseva T, Monola J, Salli E, Koivuniemi R, Savolainen S, Valkonen S, Yliperttula M. (2022)

Abstract

"Nanofibrillated cellulose (NFC) hydrogel is a versatile biomaterial suitable, for example, for three-dimensional (3D) cell spheroid culturing, drug delivery, and wound treatment. By freeze-drying NFC hydrogel, highly porous NFC structures can be manufactured. We freeze-dried NFC hydrogel and subsequently reconstituted the samples into a variety of concentrations of NFC fibers, which resulted in different stiffness of the material, i.e., different mechanical cues. After the successful freeze-drying and reconstitution, we showed that freeze-dried NFC hydrogel can be used for one-step 3D cell spheroid culturing of primary mesenchymal stem/stromal cells, prostate cancer cells (PC3), and hepatocellular carcinoma cells (HepG2). No difference was observed in the viability or morphology between the 3D cell spheroids cultured in the freeze-dried and reconstituted NFC hydrogel and fresh NFC hydrogel. Furthermore, the 3D cultured spheroids showed stable metabolic activity and nearly 100% viability. Finally, we applied a convolutional neural network (CNN)-based automatic nuclei segmentation approach to automatically segment individual cells of 3D cultured PC3 and HepG2 spheroids. These results provide an application to culture 3D cell spheroids more readily with the NFC hydrogel and a step towards automatization of 3D cell culturing and analysis."

 

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Merivaara A, Koivunotko E, Manninen K, Kaseva T, Monola J, Salli E, Koivuniemi R, Savolainen S, Valkonen S, Yliperttula M. (2022). "Stiffness-Controlled Hydrogels for 3D Cell Culture Models". Polymers. 14(24):5530.

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