| AVS 60th International Symposium and Exhibition | |
| Biomaterial Interfaces | Monday Sessions |
| Session BI+AS+IS+NL-MoM |
| Session: | Surfaces to Control Cell Response |
| Presenter: | F.A. Simoes, University of Nottingham, UK |
| Authors: | F.A. Simoes, University of Nottingham, UK C. Alexander, University of Nottingham, UK G. Mantovani, University of Nottingham, UK L. Buttery, University of Nottingham, UK M.R. Alexander, University of Nottingham, UK |
| Correspondent: | Click to Email |
Stem cells have the ability to repair, replace or regenerate tissues. As a result their potential for regenerative medicine is vast. The processing of cells for therapeutic use and clinical diagnostics will rely on cell sorting steps to ensure a homogeneous population is obtained.1
Several techniques exist to achieve this, which rely on the physical properties of cells but tend to provide poor specificity.2-4 Fluorescence Activated Cell Sorting (FACS) and Magnetic-Activated Cell Sorting (MACS) rely on specific biomarkers. However cells require labelling and label removal steps, which can affect the phenotype.5
There is a need for a fully synthetic, inexpensive, label-free separation system, capable of sorting cells with minimum manipulation. In order to generate robust surfaces for such a system, we have developed a method to immobilize thiol-functionalised materials to a polymer substrate using thiol-ene “click” chemistry in a high throughput format. Microarrays of these functionalised polymers comprising of 6 replicates, are fabricated using pin printing to generate a combinatorial library of materials. A mixture of differentiated cells derived from mouse embryoid bodies are then seeded onto the arrays.
Immunohistochemistry techniques are employed to track the differentiation of cells into different lineages, thus enabling the visualisation of multiple cell lines. These techniques also allow for the high throughput quantification of attachment by the means of automatic fluorescence microscopy.
Surface characterisation of the “click” immobilization procedure is performed by X-Ray Photoelectron Spectroscopy. In contrast the characterisation of microarrayed materials is performed using Time of Flight - Secondary ion Mass Spectrometry, which is followed by the ranking of materials using Partial Least Square (PLS) regression analysis. This process allows for the correlation of cell attachment with key molecular ions generated from each material by mass spectrometry.
Successful materials that selectively induce cell attachment are identified and investigated further. This is the first step in the generation of new surface-based devices that have the capacity to be fully synthetic, selective, inexpensive and disposable.6
1. McIntyre C. et al., Bioprocess International, 2010, 44-53.
2. Chabert M. and Viovy J., PNAS, 2008, 105, 3191-3196.
3. Shim S. et al., Integrative Biology, 2011, 3, 850-862.
4. Kose A. R. et al., PNAS, 2009, 106, 21478-21483.
5. Bulte J. W. M. et al., Blood, 2004, 104, 3410-3413.
6. Singh A. et al., Nature Methods, 2013, 10, 438-444