(277h) Engineered Cellulose-Based Cell Culture Platforms to Improve Human Health

Two-dimensional (2D) in vitro cell cultures do not mimic the microenvironment in the native tissues.Currently, there are several systems that are used to study cell cultures in 3D; however, these systems use complex instrumentation and expensive set-ups, require long and tedious optimization procedures, and do not yield in physiologically relevant structures. Other drawbacks of these constructs include heterogeneous distribution of cells, and potential mass transport limitations. Collectively, it is challenging to fabricate biocompatible constructs in cm-scale. We developed simple cellulose-based cell culture platforms that can be used for a wide range of applications for mammalian cells, bacteria, fungi, and plants cells. These new systems are simple, tunable, and low-cost.

We used cellulose-based materials as 3D scaffolds to support cells and hydrogels. We grew different types of cells including connective tissue cells, osteoblasts, neutrophils, gut cells, tumor cells, stem cells, plant cells, fungi, or bacteria in cellulose-based materials. Subsequently, we characterized the cell behavior by using viability assays, metabolic activity assays, immunocytochemistry, mechanical tests, X-rays scanning, and high-resolution imaging.

In this study, we successfully fabricated cellulose-based scaffolds to (i) investigate migration of primary human cells (tumor fibroblasts and lung cancer cells), (ii) induce template-guided biomineralization, (iii) generate 3D osteoblast cultures that are amenable to high-throughput sample preparation and analysis, (iv) form and control gradients of oxygen and biological molecules, and (v) develop new co-culture systems in multi-layered stacked cultures. In addition to primary cells and mammalian cell lines, we also cultured bacteria, fungi, and plant cells in cellulose-based materials. These cell cultures in cellulose resulted in high cell viability and high metabolic activity, and enabled differentiation of cells. In these experiments, we did not observe mass transport limitations to the cells. Our results have indicated that the cellulose-based materials allow for patterning and easy recovery of cells, can adapt modular configurations, and can provide physiologically relevant tissue models. The scaffolds we fabricated from thin layers of cellulose also yielded in free-standing structures in different scales (micron- to cm-).

To sum up, we developed new cellulose-based cell culture platforms to provide multicellular and compartmentalized tissue-mimetics for clinical applications. To overcome the limitations of the traditional tissue models, we developed a novel layer-by-layer approach to assemble tissue-like structures from cellulose-based low-cost materials. This strategy offers unique opportunities such as understanding fundamental biology, developing disease models for personalized medicine, and assembling different organs together.