Key Characterization and Synthesis Tools for Defined Polymeric Hydrogels to Guide Cell Fate

点击翻译按钮获取中文摘要

The primary objective is to examine the role of the extracellular matrix on cell fate and specifically to design a series of polymeric hydrogels and investigate their impact on cell differentiation, proliferation and guidance. Current methods of growing cells in culture provide limited information due to lack of biological cues. This leads to significant false positives (and likely just as many false negatives) in drug screening. To overcome this deficit we propose to synthesize biomimetic hydrogels for 3-dimensional cell culture and subsequent drug screening. Moreover, many cells cannot be cultured by traditional 2-dimensional techniques, requiring xenograft mouse models, which are expensive and laborious. Our 3D hydrogel strategy promises to overcome these deficits and will enable the culture of numerous cell types. Another key challenge that we aim to overcome with a defined biomimetic ECM is cell survival after transplantation. Traditional methods use saline for cell delivery with the consequence that nearly all (if not all) of the cells die. Delivering the cells in a defined hydrogel with appropriate factors will result in improved survival. The proposed equipment is key to our success. Peptides form key attachment sites for the cells through specific integrin interactions, necessitating a peptide synthesizer with which we can synthesize customized, functional peptides. The proposed peptide synthesizer will use 90% less solvent and take minutes instead of hours for coupling chemistry, making this system more efficient and environmentally friendly. The gel count is a critical instrument with which to interrogate our cells cultured in 3D. There is only one other GelCount in Toronto (at the Hospital for Sick Children), which is unpractical for daily use given the distance between our facilities and the consequent effect this distance would have on our cultured cells. The upgrade to the mechanical tester is critical to our optimized use of the mechanical tester, which is specifically designed to measure the mechanical properties of weak materials, such as our hydrogels. This relatively small investment will provide great utility to our advancement of knowledge. The incubator is key to the successful functioning of any cell culture facility. There are over 120 highly qualified personnel in our combined laboratories who will benefit from the proposed equipment. It will provide them with the tools required to design and evaluate novel biomaterials for use in biomedical engineering applications.