Homogeneous seeding of mesenchymal stem cells into nonwoven fabric for tissue engineering.

Mesenchymal stem cells (MSCs) were isolated from the bone marrow of rats and seeded into a nonwoven fabric of polyethylene terephtalate (PET) by agitation and static methods. MSC attachment was investigated in terms of the number of cells attached to the fabric, their distribution inside the fabric, and cell damage. The number of MSCs attached was greater for the agitation seeding method than for the static seeding method. The higher the rotating speed in the agitation seeding method, the greater the number of cells attached. When the cell suspension was seeded into the fabric in culture medium volumes of 50 and 200 microL per well of the culture plate or per culture tube, the best cell attachment was observed for the tube culture group at the larger volume. These cells attached more homogeneously throughout the fabric in greater numbers than was the case for the other culture groups. It is possible that agitation of the cell suspension allows cells to infiltrate uniformly inside the fabric, resulting in a homogeneous distribution of the cells in the fabric. A biochemical study revealed that neither the agitation nor static seeding method damaged cells, irrespective of the medium volume and the type of culture vessel. We conclude that the agitation seeding method is a promising method by which to formulate a homogeneous construct of fabric and MSCs.