| AVS 59th Annual International Symposium and Exhibition | |
| Biomaterial Interfaces | Monday Sessions |
| Session BI-MoA |
| Session: | Cell-Surface Interactions: High Throughput Methodologies |
| Presenter: | A.D. Celiz, Univeristy of Nottingham, UK |
| Authors: | A.D. Celiz, Univeristy of Nottingham, UK M. Mahlstedt, University of Nottingham, UK A.L. Hook, University of Nottingham, UK D.J. Scurr, University of Nottingham, UK D.G. Anderson, Massachusetts Institute of Technology R. Langer, Massachusetts Institute of Technology D.A. Barrett, University of Nottingham, UK C. Denning, University of Nottingham, UK L. Young, University of Nottingham, UK M.C. Davies, University of Nottingham, UK M.R. Alexander, University of Nottingham, UK |
| Correspondent: | Click to Email |
Numerous regenerative medicine procedures are already in clinical trials or in the pipeline and if these succeed and reach the clinic, stem cell factories will be needed to meet demand for the billions of cells required per intervention. Current protocols for stem cell culture employ poorly defined biological substrates such as MatrigelTM, and/or non-human feeder cell layers which exhibit batch-to-batch variability, and are a potential source of pathogens. More recently, recombinant protein surfaces have been successfully employed, but these are not always cost effective for high-throughput culture methods. Consequently, there is significant research into xeno-free culture alternatives that use fully defined culture media and synthetic substrates. To discover materials for application as substrates, we have used high-throughput polymer arrays with which to fabricate chemically defined and scalable stem cell culture systems. A combinatorial chemical polymer library can be synthesized as a microarray on-slide allowing cell response of hundreds of different materials on a single glass slide, enabling hit materials to be identified in an automated manner [1]. High-throughput surface characterization (HT-SC) can be performed on the polymer microarray to (i) rapidly evaluate the surface chemistry of each individual spot and (ii) identify surface structure-property relationships in the complex and large datasets generated. (2)
In this study, a polymer library substantially increased in number and diversity compared with previous studies is prepared by printing (meth)acrylate monomers in pair-wise combination on a polyHEMA coated slide. HT-SC is carried out using time-of-flight secondary-ion mass spectrometry, X-ray photoelectron spectroscopy, atomic force microscopy and water contact angle to characterize all polymer surfaces. Human pluripotent stem cell (hPSC) attachment is probed using automated fluorescence microcopy. This suite of HT-SC techniques has allowed the identification of ‘hit’ materials that support the expansion and proliferation of hPSCs. Human embryonic stem cell lines (HUES7 and H9) are screened for attachment in 3 different media; mouse embryonic fibroblast-conditioned media (MEF-CM), StemPro® and mTeSR®1 without any protein preconditioning to provide an insight into the impact of material surface properties on cellular interactions. The hits identified represent novel platforms for hPSCs culture without the need for any protein preconditioning and show great potential for development into future synthetic culture systems.
[1] Anderson, D. G. et al. Nat. Biotechnol. 22, 863 - 866 (2004).
[2] Mei, Y. et al. Nat. Mater. 9, 768 – 778 (2010).