Paper BI+AS+IS+NL-MoM10
What Makes the Heart Grow Fonder? Chemically Diverse Polyacrylate and Polyacrylamide Surfaces for Human Cardiomyocyte Culture and Their Effect on Phenotype
Monday, October 28, 2013, 11:20 am, Room 201 B
| Session: |
Surfaces to Control Cell Response |
| Presenter: |
A.K. Patel, University of Nottingham, UK |
| Authors: |
A.K. Patel, University of Nottingham, UK
D.G. Anderson, Massachusetts Institute of Technology
R. Langer, Massachusetts Institute of Technology
M.C. Davies, University of Nottingham, UK
M.R. Alexander, University of Nottingham, UK
C. Denning, University of Nottingham, UK
|
| Correspondent: |
Click to Email |
Human pluripotent stem cell (hPSC) derived cardiomyocytes hold the potential to strengthen pharmaceutical toxicity testing and to provide disease models for development of treatment targets1. The maturation and maintenance of the cardiomyocyte phenotype may be controlled by the manipulation of the substrate supporting the cells2. However, the surfaces currently in use still fall short of producing cardiomyocytes of adult maturity. Standard culture-ware requires coating with biological substrates such as fibronectin which can be expensive and subject to poor reproducibility due to batch variation. We are exploring an alternative, combinatorial materials high throughput screening approach3 to identify novel materials that can improve cardiomyocyte culture. Polymer microarrays comprising of 6 replicates of 116 acrylates and acrylamides are fabricated using contact printing. Cardiomyocytes derived from the HUES7 human stem cell line are seeded onto the arrays. Immunostaining of nuclei (DAPI) and the cardiomyocyte specific motor protein, sarcomeric alpha actinin is performed to visually estimate cell function and maturity and enable quantification of cell attachment in a high throughput manner using automated fluorescence microscopy and image analysis software. Surface characterisation of the arrays is performed using time of flight secondary ion mass spectrometry. Partial least squares (PLS) regression analysis allows for correlation of cell attachment with key molecular ions identified from mass spectrometry4. Successful monomers that permit cardiomyocyte attachment, spreading and contraction are identified from the first generation homopolymer microarray and are mixed pair-wise to form second generation microarrays. This diverse library of copolymers enables unique combinations of chemical moieties to be investigated. Hit monomers and combinations identified to be synergistic can be analysed for their effect on cardiomyocyte function including electrophysiology measured by patch clamping, myofibril alignment and gene expression.
The lead materials generated by this approach are the first step in a discovery process for novel synthetic biomaterials capable of enhancing the culture of cardiomyocytes to move towards more reproducible, economical and defined conditions.
References: 1. Matsa E. et al. European Heart Journal. 2011;32(8):952-62
2. Engler A. et al.The Journal of Cell Biology. 2004;166(6):877–887
3. Hook A. et al. Biomaterials. 2010;31(2):187–198
4. Yang J. et al.Biomaterials. 2010;31(34): 8827–8838