3D cell culture and assay automation demonstrated with a hydrogel derived from wood

Abstract:

Introduction
In the pursuit of in vitro cell models with improved functionality, materials and methods for three-dimensional (3D) cell culture are a key requirement. GrowDex®, a novel nanofibrillar cellulose (NFC) hydrogel derived from wood, has been shown to provide an effective support matrix aiding the transition from 2D to 3D cellular models.

Results
Studies with GrowDex have shown it to be an effective support matrix for a variety of applications. Hepatic cells form 3D spheroids with polarized structures and better functionality of the metabolic enzymes in GrowDex when compared to traditional 2D culture on plastic, and long term culture of primary hepatocytes enables repeated dose drug toxicity testing. Proliferation of embryonic stem cells and induced pluripotent stem cells without feeder cells has been successfully accomplished in GrowDex, as well as the proliferation and differentiation of mesenchymal stem cells. Grown 3D cell structures can be collected from GrowDex for detailed downstream analysis of the cells by cellulase enzyme treatment. The shear-thinning property of GrowDex permits its use with automated dispensing systems at ambient temperature in high throughput screening (HTS) –related 3D cell culture applications.

Conclusions
GrowDex is animal-free, ready-to-use hydrogel with shear-thinning properties that enable injecting and pipetting of the material. It is biocompatible with human cells and tissues and does not contain any animal or human derived material. 3D cell growth is made possible as GrowDex physically resembles extracellular matrix (ECM) allowing free diffusion of small molecules, such as nutrients and oxygen. GrowDex can be completely degraded to soluble glucose by enzyme treatment, whilst the 3D cellular structure is retained with no adverse impact on cells observed.

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