| AVS 60th International Symposium and Exhibition | |
| Biomaterial Interfaces | Wednesday Sessions |
| Session BI-WeM |
| Session: | Cell-Surface Interactions |
| Presenter: | A.D. Celiz, University of Nottingham, UK |
| Authors: | A.D. Celiz, University of Nottingham, UK J.G.W. Smith, University of Nottingham, UK A.K. Patel, University of Nottingham, UK R. Langer, Massachusetts Institute of Technology D.G. Anderson, Massachusetts Institute of Technology D.A. Barrett, University of Nottingham, UK L.E. Young, University of Nottingham, UK C. Denning, University of Nottingham, UK M.C. Davies, University of Nottingham, UK M.R. Alexander, University of Nottingham, UK |
| Correspondent: | Click to Email |
A key hurdle in translating stem cell therapies from research to industrial scale and clinical application is to produce the necessary numbers of cells. For example, a major heart attack causes loss of 1 billion cardiomyocytes and similar cell numbers are lost during progression of other conditions such as multiple sclerosis and diabetes. To meet the demand for such high cell numbers, a defined growth substrate free of animal-derived components is desirable. To this end, we have employed polymer microarrays to screen for human pluripotent stem cell (hPSC) attachment and phenotype on polymers in a high-throughput manner. Polymer microarrays enable a large combinatorial chemical space to be interrogated on a single glass slide. Furthermore, since monomers can be robotically printed and polymerized on the slide via UV photo polymerization, rapid evolution of large numbers of polymers is facilitated. ‘Hit’ materials identified from the initial screen can be taken forward to a second generation array and mixed in a combinatorial manner to test hypotheses formed from the first generation array and this iteration continues until scaled up to plastic ware for automated culture protocols to achieve long-term expansion of hPSCs.
We have screened over 140 acrylate and acrylamide homopolymers in an array format for hPSC attachment in mouse embryonic fibroblast (MEF) conditioned medium and defined media including StemPro® and mTeSR™1 after 24 hours in culture. Hit materials were then mixed to produce a second generation array of over 500 unique copolymers to optimise cell attachment. Polymer microarrays were characterized using time of flight secondary-ion mass spectrometry (ToF-SIMS), X-ray photoelectron spectroscopy (XPS), atomic force microscopy (AFM) and water contact angle (WCA) measurements. Multivariate analysis (MVA) was used to successfully predict material wettability and cell performance from the ToF SIMS data. Cell attachment was identified using DAPI (nuclei) staining and maintenance of pluripotency was confirmed by OCT-4 staining and imaged using automated fluorescence microscopy. This approach to materials discovery will provide a defined, synthetic growth substrate for hPSC culture that is amenable to scale up for industrial application and is a step toward xeno-free hPSC culture conditions necessary for clinical application.