Development the Technique for the Preparation and Characterization of Reconstructed Human Epidermis (RHE)

Abstract

Reconstructed Human Epidermis (RHE) is an artificial epidermis made in such a way that it resembles human skin, and can be used for the identification of irritant chemicals, especially for cosmetic and topical medicinal products. Currently the new RHE is produced by European and American countries, whose skin physiology is very different from Indonesia. Based on this, the Center for Research on Drugs and Food, NADFC of Republic of Indonesia took the initiative to develop the reconstruction of keratinocyte, melanocytes and fibroblasts cells into RHE adapted to the anatomical and physiological functions of the skin of Indonesians. RHE is made from an epidermal layer composed of keratinocyte and melanocytes cells that are reconstructed with a dermis layer composed of fibroblast and collagen cells. Keratinocyte, melanocytes and fibroblasts cells are cultured on suitable mediums by adding a suitable growth medium. To find out that RHE has been successfully reconstructed, measured percentage of cell life, made histology preparation to see the existence of cell nucleus, and conducted Immunohistochemical examination to see existence of integration (bond) between antigen. From the research results can be seen that keratinocyte cells grown on culture medium Keratinocyte SFM (IX) with rEGF supplements; melanocyte cells grown on Melanocyte 254 culture medium with HMGS supplementation; and fibroblast cells grown on Fibroblast M 106 culture medium with LSGS supplementation. The percentage of epidermal cell life grew well in the planting of 10 Â 10 4 cells /mL keratinocytes and 0.25 Â 10 4 cells /mL of melanocyte cells and survived until the 11 th day with live cell percentage of 93.45%. In making preparation for histology with HE staining, there is a cell life in RHE tissue. Used Immunohistochemical (IHC) examination using cytokeratin 10 antibody marker to view physiological function of epidermal tissue.

Classic and Current Opinions in Human Organ and Tissue Transplantation

“…A few examples of xenograft include grafting human keratinocytes onto non-human cells (e.g., mice) and then using "ZenSkin" (Reconstructed Human Epidermis) construction as a model for human skin physiology. ZenSkin has applications in pre-clinical and R&D for evaluating how a topical product will affect the human skin [29,30]. Other examples include transfusing non-human blood into human patients and skin grafts from non-humans.…”