A cell bank system for cultured epithelial sheet transplantation

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1)Development of preservation and transportation method for biopsied skin and cultured epithelial sheet. For incorporating cultured epithelial sheet technique into Japanese medical care system, it is necessary to establish culture center for supplying cultured epithelial sheet. We examined the best medium to keep biopsied skins for 4 days under refrigerated condition. We found that L-15 medium is suitable for this purpose. Next, we studied how long cultured epithelial sheet could be stored in favorable condition after detachment from culture plate. Cultured epithelial sheet could be preserved in good condition under refrigerated condition with Hanks solution for 12 hours, but not foe 24 hours. This result led us to develop a new method to preserve cultured epithelial sheet under frozen condition. We found that L-15 medium containing polyethylene glycol 600 is the best medium for this purpose. Cultured epithelial sheet could be frozen and stored in ultra-deep freezer (-150ﾟC) for up to … More 3 months. Furthermore, storage in dry ice for 72 hours did not cause any unfavorable damage to cultured epiderm sheet. We also confirmed that cultured epithelial cells could be stored in liquid nitrogen for over 20 years theoretically. 2)Assessment of clinical application of cultured epithelial sheet. First, we performed the compared analysis for the effectiveness of frozen and non-frozen cultured epithelial sheet. There were no significant difference between both of them. Based on this result, we used frozen autograft cultured epithelial sheet in general. We tried clinical application of cultured epithelial sheet mainly for recurrent leg ulcer, epidermolysis bullosa and sever burn. The marked improvement was observed in epidermolysis bullosa, in which durati on of ulcer epihtelization was markedly shortened and blister formation due to mechanical force was decreased in transplanted lesions. In leg ulcer, repetitive transplantation of cultured epithelial sheet was performed using frozen-stored epidermal keratinocytes. Thus, the possibility of cell bank system in cultured epithelial sheet transplantation was suggested. Less