Efficient transfer of human adipose-derived stem cells by chitosan/gelatin blend films

Adipose-derived stem cells (ASCs) are a potential source of abundant mesenchymal stem cells and represent a promising cell-based therapy for tissue damage or degeneration conditions. Previous investigations have demonstrated enhanced therapeutic effects of ASCs in a three-dimensional spheroid culture formulation. In this study, we hypothesize that a composite membrane made of chitosan/gelatin (C/G) is beneficial to facilitate transfer of human ASCs in spheroids. Increasing chitosan content within the blends enhanced the mechanical properties of the sample, including tensile strength and elongation-at-break ratio. Although ASC spheroids developed shortly after seeding on pure chitosan films, increasing gelatin proportion in the C/G blends promoted cell adhesion onto the membranes. We also found that ASCs did not proliferate on chitosan films, but C/G blends of different ratios supported ASC proliferation in the first 4 days of culture. However, ASCs on all C/G blends started to detach from the films to form spheroids after day 4, while ASCs on pure gelatin films remained attached and continued to grow. Gradual gelatin release from the C/G blend films, leading to enriched chitosan content in the blends, probably encouraged ASC detachment and spheroid formation. We placed porous collagen matrix on ASC-seeded C/G blends to simulate the application of ASC-seeded C/G films onto injured tissue and found that a C/G film composed of 75% chitosan could facilitate significantly more cell transfer into the overlying collagen sponge. Therefore, a blend film containing 75% chitosan and 25% gelatin showed promising results to serve as a biomaterial for human ASC-based cell therapy.