抄録
To study the effects cosmetic materials on normal human keratinocyte (NHK) differentiation, we investigated the evaluation method of differentiation in NHK.
Involucrin is a precurser protein of detergent-insoluble cornified envelope and a marker of terminal differentiation of epidermal keratinocytes. To quantify differentiation of NHK, the population of involucrin-positive cells was estimated by immunofluorescent staining using antiinvolucrin antibody and flow cytometry. NHK were cultured under three conditions: 0.1mM Ca2+, 1.8mM Ca2+, and 1.8mM Ca2+ with 10% fetal calf serum (FCS). The population of in volucrin-positive cells increased from 8.0% to 22.1% (2.8 fold) in case of 1.8mM Ca2+, and to 37.1% (4.6 fold) with addition of FCS. In addition, the rate of involucrin-positive cells was correlated significantly with the production rate of cornified envelope under the same condition.
The cytofluorographic analysis of involucrin expression made it possible to determine the number of differentiated cells precisely and reliably in a large number of samples. Thus, the method described here seems to be a useful method for evaluation of keratinocyte differentiation.
