Development the Technique for the Preparation and Characterization of Reconstructed Human Epidermis (RHE)
Herlina B. Setijanti, Herlina
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 � 104 cells /mL keratinocytes and 0.25 � 104 cells /mL of melanocyte cells
and survived until the 11th 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.