Abstract
Cell sheets have been studied for the purpose of regenerating dysfunctional tissues and it has also been reported that systemic transplantation of autologous cultured epidermis was actually successful. However, there are few simple methods for testing activity after cell sheet preparation. In this study, we aimed to reproduce the state of sunburn on a part of normal human epidermal keratinocyte sheet by UV irradiation and to evaluate the activity using Bio-LSI. The evaluation examined the respiratory bursts in immune cells by monitoring in real-time, the O2 consumption of the epidermal cell sheet. The cell sheet culture kit Lab Cyte EPI-KIT (Japan Tissue Engineering Co., Ltd.) was cultured for 18 days and were cut into a circle of about 5 mm in diameter. The cell sheet was then covered with a mask leaving 1mm × 1mm open surface for irradiation with UV light (365 nm wavelength and an illuminance of 300 mW/cm2 for 10 seconds). Following UV irradiation, it was installed in the measuring device promptly. The experimental conditions were as mentioned below: measurement solution: 11.4 mM glucose in PBS; W.E.: Pt electrode (d= 40 um) ×400 point; R.E.: Ag/AgCl electrode; C.E.: Pt wire; applied voltage: -0.5 V vs. Ag/AgCl and sampling time: 500 ms. In Fig. 3, it can be seen that the area of cell sheet illuminated with UV irradiation shown lower oxygen concentration (reflected by black area) while non-irradiated parts show higher oxygen level. It is due to the fact that UV exposed cells suddenly consumed oxygen.
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