Introduction

Cultured stratified epithelial cell sheets have been clinically utilized as transplantable grafts for the regeneration of epithelial tissues, such as the esophagus, cornea, skin, and intraoral cavity. These cell sheets are expected to gain widespread use as regenerative medicine products and save many patients. For this purpose, establishing and disseminating the stale protocol of fabricating the cell sheet is crucial. The fabrication of cultured stratified epithelial cell sheets consists of many important steps, and since the patients’ epithelial cell conditions vary widely and are sometimes unstable, the qualities of the epithelial cell grafts are likewise potentially unstable. Therefore, in this paper, we report the stable protocol for fabrication of the transplantable cell sheet particularly from patient-derived oral mucosal tissues.

Methods

Serum extracted from blood and buccal mucosal tissue were collected in Nagasaki University and transported to Tokyo Women's Medical University. Oral mucosal epithelial cells were collected by minimum trypsin method, and this treatment was studied whether to be a critical procedure. After 14 days cultivation, cultured cells were examined whether to be transplantable as cell sheets.

Results

We successfully transported buccal mucosal tissue and serum without damage and contamination. Oral mucosal epithelial cells were collected with high viability by minimum trypsin method. Finally, we succeeded to stably fabricate oral mucosal epithelial cell sheets in all 10 patients.

Conclusions

We established a stable protocol for the fabrication of human oral mucosal epithelial cell sheets and their transportation in clinical settings in this study. These methodologies could also be basis for transplantation therapy using cultured cell sheets of various types other than oral mucosal epithelial cell and will contribute largely to the future development of regenerative medicine.

中文翻译：

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用于制造用于临床应用的可移植人口腔粘膜上皮细胞片的稳定方案。

介绍

培养的分层上皮细胞片已在临床上用作可移植的移植物，用于上皮组织的再生，例如食道、角膜、皮肤和口腔。这些细胞片有望作为再生医学产品得到广泛应用，并拯救许多患者。为此，建立和传播制造细胞表的陈旧协议至关重要。培养的分层上皮细胞片的制造包括许多重要步骤，并且由于患者的上皮细胞状况差异很大并且有时不稳定，因此上皮细胞移植物的质量同样可能不稳定。因此，在本文中，我们报告了用于制造可移植细胞片的稳定方案，特别是来自患者衍生的口腔粘膜组织。

方法

在长崎大学收集从血液和颊粘膜组织中提取的血清并运送到东京女子医科大学。用最小胰蛋白酶法收集口腔黏膜上皮细胞，研究这种处理是否是关键程序。培养14天后，检查培养的细胞是否可移植为细胞片。

结果

我们成功地运输了颊粘膜组织和血清，没有损伤和污染。通过最小胰蛋白酶法收集具有高活力的口腔粘膜上皮细胞。最后，我们成功地在所有 10 名患者中稳定地制造了口腔粘膜上皮细胞片。

结论

在本研究中，我们建立了一个稳定的方案，用于制造人口腔粘膜上皮细胞片及其在临床环境中的运输。这些方法也可以作为使用除口腔粘膜上皮细胞以外的各种类型的培养细胞片进行移植治疗的基础，并将在很大程度上促进再生医学的未来发展。