Preparation and Application of New Type Matrix for Aligned Cell Sheet Engineering Use

In recent years, we developed a system using γ-PGA as base and disulfide bonds cleavage as the critical point of cell sheet detach system. The novel cell sheet detaching system showed much higher cell adhesion and cell activity. Although many literatures demonstrate that cell sheet engineering is suitable culture technology for squamous epithelial tissue, the high regularity stromal tissue has been a major challenge for tissue engineering. The technical concept of cell sheet engineering as the basis of a surficial modified PVDF membrane, when the intermediate linkage was then cystine upper γ-PGA as a cell culture carrier, and use a disulfide bond cleaving mechanism key steps to achieve the purpose of the cell sheet culture systems. The novel cell sheet engineering technology and it application has been applied to the Taiwan and U.S. Patent Application, and the detailed protocols also was advance technology transferred to cord blood biotech industry to culture mesenchymal stem cells for clinical application research. However, mesoderm derivative tissues is complex and with high regularity, so its culture has long been considered to be very utility and challenging. The NSC grant of this year is an extension of cell sheet engineering. Therefore, we will develop an aligned cell sheet engineering system within three years which base on a transparent PVDF membrane with various patterns, and attempt to culture complex and highly regular mesodermal tissues such as corneal stromal tissue or corneal endothelial tissue. Briefly, these various patterns on PVDF membrane will be fabricated by soft-lithography technology or dip-pen nanolithography technology, and then disulfide bond-containing amino acid and biopolymer will be led onto plasma activated PVDF membrane surface. Overall, the grant will establish an aligned cell sheet engineering technology platform to culture various cell sheet with different regularity can be used to reconstruct stromal tissue for future clinical applications.