Surface modification of closed plastic bags for adherent cell cultivation

In modern medicine human mesenchymal stem cells are becoming increasingly important. However, a successful cultivation of this type of cells is only possible under very specific conditions. Of great importance, for instance, are the absence of contaminants such as foreign microbiological organisms, i.e., sterility, and the chemical functionalization of the ground on which the cells are grown. As cultivation of these cells makes high demands, a new procedure for cell cultivation has been developed in which closed plastic bags are used. For adherent cell growth chemical functional groups have to be introduced on the inner surface of the plastic bag. This can be achieved by a new, atmospheric-pressure plasma-based method presented in this paper. The method which was developed jointly by the Fraunhofer IST and the Helmholtz HZI can be implemented in automated equipment as is also shown in this contribution. Plasma process gases used include helium or helium-based gas mixtures (He + N2 + H2) and vapors of suitable film-forming agents or precursors such as APTMS, DACH, and TMOS in helium. The effect of plasma treatment is investigated by FTIR-ATR spectroscopy as well as surface tension determination based on contact angle measurements and XPS. Plasma treatment in nominally pure helium increases the surface tension of the polymer foil due to the presence of oxygen traces in the gas and oxygen diffusing through the gas-permeable foil, respectively, reacting with surface radical centers formed during contact with the discharge. Primary amino groups are obtained on the inner surface by treatment in mixtures with nitrogen and hydrogen albeit their amount is comparably small due to diffusion of oxygen through the gas-permeable bag, interfering with the plasma-amination process. Surface modifications introducing amino groups on the inner surface turned out to be most efficient in the promotion of cell growth

Surface modification of closed plastic bags for adherent cell cultivation

In modern human mesenchymal stem cells are gaining increasing importance. Usein helium-based gas mixtures (He+N2+H2) as well as helium enriched with suitable film- forming agents the inner surface of polymer bags can be modified with a plasma process development at Fraunhofer IST. The effect of plasma treatment in investigated by FTIR-ATR spectroscopy as well as surface tension determination based on contact angle measurements. Plasma treatment in nominally pure helium increases the surface tension of the polymer foil due to the presence of oxygen traces in the gas and oxygen diffusing through the gas-permeable foil, resp., reacting with surface radical centres formed in the discharge. For the same reason the area density of primary amino groups on the surface obtained by treatment in mixtures with nitrogen is comparably small. With regard to their applicability in cell cultivation three different coatings (bases on APTMS, DACH and TMOS) were tested. It is shown that primary amino groups play an important role in cell cultivation of adherent growing bone marrow mesenchymal stem cells