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A cell bank system for cultured epithelial sheet transplantation

用于培养上皮片移植的细胞库系统

基本信息

批准号：
06454317
负责人：
YOSHIKAWA Kunihiko
金额：
$ 4.1万
依托单位：
Osaka University
依托单位国家：
日本
项目类别：
Grant-in-Aid for General Scientific Research (B)
财政年份：
1994
资助国家：
日本
起止时间：
1994 至 1995
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-06454317/
关键词：
Cultured epithelial sheet Transplantation Ultra-deep freezer Epiderm cells Leg ulcer Epidermolysis bullosa Burn Cell bank 表皮ケラチノサイト培養表皮シート移植細胞バンク培養センター L-15

项目摘要

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

1）活检皮肤和培养上皮片的保存和运输方法的开发。为了将培养上皮片技术纳入日本的医疗体系，有必要建立提供培养上皮片的培养中心。我们研究了将活检皮肤在冷藏条件下保存 4 天的最佳介质。我们发现 L-15 培养基适合此目的。接下来，我们研究了培养的上皮片从培养板脱离后可以在良好的条件下保存多长时间。培养的上皮片在Hanks溶液冷藏条件下可以保存12小时，但不能保存24小时。这一结果促使我们开发了一种在冷冻条件下保存培养的上皮片的新方法。我们发现含有聚乙二醇 600 的 L-15 培养基是用于此目的的最佳培养基。培养的上皮片可以冷冻并在超深冰箱（-150°C）中保存长达 3 个月。此外，在干冰中储存72小时不会对培养的表皮片造成任何不利的损害。我们还证实，培养的上皮细胞理论上可以在液氮中保存20年以上。 2)培养上皮片的临床应用评价。首先，我们对冷冻和非冷冻培养上皮片的有效性进行了比较分析。两者之间没有显着差异。基于这一结果，我们一般使用冷冻自体移植培养的上皮片。我们尝试了培养上皮片的临床应用，主要用于治疗复发性腿部溃疡、大疱性表皮松解症和严重烧伤。在大疱性表皮松解症中观察到明显的改善，其中溃疡上皮形成的持续时间显着缩短，并且移植病灶中由于机械力导致的水疱形成减少。在腿部溃疡中，使用冷冻保存的表皮角质形成细胞重复移植培养的上皮片。由此，提出了细胞库系统在培养上皮片移植中的可能性。较少的