Cell delivery with genipin cross-linked gelatin microcarrier in liver tissue engineeing

Tissue engineering has shown vast advancement in alleviating the urgent need for donor organs. Three-dimensional scaffolding concept in tissue engineering holds a promising approach in developing liver tissue to replace or restore the functions of degenerated liver. Herein, an optimised fabricated genpin cross-linked gelatin microspheres of two size-ranges that provide an injectable method for HepG2 cell delivery was introduced. Optimised conditions of subsequent incubation temperature at 37oC and ethanol concentration of 90% were achieved to allow the synthesis of uniform cross-linked and well dispersed microspheres. Fluorescent live/dead staining illustrated a metabolically active, viable cell aggregation on both sizes of microspheres but greater cell aggregates formations were seen on large spheres which was confirmed by SEM observations. As tested by WST-1 assay, cells cultured on large spheres were proven to have better proliferation rate than in small spheres. Maintenance of cell functionalities throughout the culture period was confirmed by albumin and urea secretions. Collectively, the results have shown a better performance of large spheres as scaffold in cell delivery, where more viable cells and enhanced functionalities were observed. Proposed reason of such observations might be due to substrate curvature, which affects the energy required for microtubules bending in anchorage dependent HepG2 cells for adhesion onto substrate. Microtubules in HepG2 cells, responsible for many diverse liver-specific functions, when disrupted may cause undesired effects on the subsequent cellular processes and activities. Besides raising the awareness on the possible effects of scaffold structure on cells behaviours, this study had assured the superior usage of microspheres as microcarriers in cell delivery purposes in the field of tissue engineering research.Bachelor of Engineering (Chemical and Biomolecular Engineering