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Cell proliferation and differentiation on synthetic nanofibrillar surface

M.A. Lodhi, G.W. Opperman and S.M. Larkin
SurModics, Inc., US

Keywords:
nanofiber, synthetic cell culture matrix, PC12, ES-D3

Abstract:
In recent years there has been a growing concern among the cell biologists regarding the use of flat surfaces for growing cells - a tradition that is more than a century old. Cells grown on the flat surfaces, such as polystyrene tissue culture plates, lack the morphology and physiology characteristics of such cells in vivo. Naturally cells grow on 3-dimensional fibrillar structures such as basal laminae and extra cellular matrix (ECM). Cells recognize the mechanical and chemical signals of the ECM and attach to proliferate, communicate, differentiate, migrate and do all sort of normal functions. The lack of such microenvironment results into abnormal cellular morphology and functioning of the cells. Proteins extracted from ECM have been used on flat surfaces to overcome this problem but the use of these proteins opens up other regulatory and usage issues. Several synthetic structures have also been tested to mimic in vivo environment, including nanofibers. We designed several experiments to test the effect of surface properties of the electrospun nanofibers chemically modified with PhotoLink® technology on growth and differentiation of PC12 and ES-D3 cells. PuraMatrix™ and Matrigel™ were used as substrates in addition to the nanofibers coated with photo polyamines. Quantitatively better cell differentiation was achieved in PC12 cells grown on coated nanofibers (60%), PuraMatrix (10%), Matrigel (5%) and uncoated polystyrene control (1%). In addition, ES-D3 stem cells attach and divide well on nanofiber surfaces coated with polyamines while remaining totipotent in the presence of FGF. With the proper change in growth conditions, D3 cells differentiated into neurons, oligodendrocytes and type I and type II astrocytes. Moreover, nanofibers proved to be an effective substrate in the long-term maintenance and growth of the cells. PC12 and ES-D3 cells grown on polyamine coated nanofibers were robust and displayed their respective morphologies even after 60 and 30 days in cultures respectively, without being passaged onto fresh substrate. In conclusion, a nanofiber-based surface provides a robust synthetic substrate for cell attachment, differentiation and long-term culture of the cells.

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