Objective

Introduction

In the pursuit of in vitro cell models that are biological relevant with improved functionality, new materials and methods for creating of three-dimensional (3D) cell culture systems are a key requirement. GrowDex^®, nanofibrillar cellulose (NFC) hydrogel which is derived from the Birch tree, has been shown to provide an effective support matrix for culturing cells in 3D. As such this hydrogel can play a vital role in simplifying the transition from 2D to 3D cellular models and furthering their use in high throughput screening (HTS).

Results

Studies have shown NFC hydrogel to be an effective support matrix for a variety of cells including primary, ES, iPS and patient derived cells. Hepatic cells have been shown to form 3D spheroids with polarized structures with better functionality of the metabolic enzymes when compared to traditional 2D culture. 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 also been successfully accomplished, as well as the proliferation and differentiation of mesenchymal stem cells, as well as culture of patient derived cells and biopsies. The shear-thinning properties of GrowDex and the fact that is not temperature sensitive nor does it require a cross-linking step, permits its use with automated dispensing systems at ambient temperature. Finally, structures grown in 3D can be collected for detailed downstream analysis by use of GrowDase™ enzyme treatment without impact to cell viability, phenotype or function.

Conclusions

GrowDex is biocompatible with both cells and tissues. It is an animal-free, ready-to-use hydrogel with shear-thinning properties that allow the hydrogel to be dispensed manually or via automated systems or injected for use in drug release studies. GrowDex physically resembles extracellular matrix (ECM) allowing free diffusion of small molecules, such as nutrients and oxygen. It can be completely degraded to soluble glucose by one-step treatment with GrowDase which preserves the 3D cellular structure with no adverse impact on cells being observed. These properties make GrowDex a suitable matrix for a culturing a wide variety cells and tissues for use in numerous 3D cell culture applications and drug discovery studies.