AN EXPERIMENTAL STUDY TO CREATE EPITHELIAL CELL SHEETS ORIENTED TO THE APPLICATION IN CASE OF SKIN TISSUE LOSS

Abstract

Amniotic membranes have been studied and applied for many years because they have anti-inflammatory, antibacterial, and antiviral properties, especially the ability to cause inadequate immune responses, which is one of the causes important for graft rejection. Therefore, amniotic membrane is one of the natural biological materials of great interest in many research and application fields. Recently, the amniotic membrane has been used as a scaffold in tissue engineering to create various types of tissue engineering grafts. Along with the continuous development of stem cell technology and the combination with tissue engineering, it is possible to develop grafts with the characteristics and functions necessary for the tissues that need regeneration. In this study, our research team used collagen membranes obtained from the amniotic membrane as a carrier for mesenchymal stem cells obtained from umbilical cord blood to create epithelial cell sheets aimed at applications in cases of skin damage.

Mesenchymal stem cells were obtained from cord blood and identified according to international standards (ISCT). These stem cells will then be transferred onto collagen membranes and epithelial sheets using a differentiation kit and specialized culture plates to create epithelial sheets. The results of epithelial sheet formation were evaluated by imaging analysis methods (H&E, TEM) and immunohistochemical staining methods (p63, keratin 1/10).

Mesenchymal stem cells satisfy the ISCT criteria for stem cell identification, such as adhesion cell characteristics, the potential to differentiate into osteoblasts, chondrocytes, and adipocytes in vitro, as well as expression showing markers such as positive markers CD73, CD90, CD105, and negative for some markers CD14, CD45. The collagen membrane obtained from the amniotic membrane has removed the cells to become an acellular membrane to serve as a cell carrier. The epithelial cell sheet showed that the cells exhibited the characteristics of epithelial cells after differentiation (positive with somes markers such as p63, ck 1/10) and formed multiple layers of cells that closely resembled the features of the epithelial layer.

The research team has successfully created epithelial cells with morphological and functional characteristics of epithelial cells. This research result has excellent application potential and is helpful in skin loss injuries such as burns and accidents when epithelial sheets are available for treatment.