Within the project, various methods were applied to study the interaction of biomatrix polymers with the surfaces of scaffolds/microcarriers. The successful application of the biomatrix was demonstrated by a proof-of-principle study using microcarriers and primary stem cells in a simplified bioreactor culture.
In the first part of the project, a layer-by-layer procedure for the deposition of the biomatrix was analyzed and optimized for stem cell growth on 2D culture surfaces. Analytical methods such as atomic force microscopy (AFM), colorimetric assays and spectrophotometry, fluorescent imaging and zeta potential measurement were applied to study the deposition of the polymers on different surfaces and critical parameters were identified. Our results indicated that charge-based absorption of polymers can be employed to coat various materials applicable for cell culture. Finally, a qualitative model for the biomatrix deposition on cell culture plastic ware was developed. The deposition model was further verified using materials with different surface charge properties.
In the second part of the project, the defined layer-by-layer procedure was transferred to 3D scaffolds and microcarriers. A model microcarrier was used to test the applicability and efficiency of the biomatrix deposition. Subsequently, various scaffolds and microcarriers made of synthetic and natural materials were successfully coated.
Finally, 3D cell culture experiments were carried out for proof-of-concept. Therefore, primary stem cells, in particular mesenchymal stromal cells, were cultured for 10 days in a shaker flask culture system. The results showed that these primary stem cells can be successfully cultured on microcarriers functionalized with the optimized layer-by-layer procedure developed in this project.
The results of this project have the potential for commercial exploitation. The deposition model and obtained data complemented a European patent application. Furthermore, existing products benefited from the development of the deposition model and optimization of coating procedure. In addition, using the optimized layer-by-layer procedure will drastically shorten the development periods of new products. In the future, the host organization plans to bring a novel microcarrier to the market that will enable improved bioreactor culture and higher cell numbers to meet the demands of cell-based industries.
