伤口愈合的生理过程是动态、连续且复杂的。目前，自体皮肤移植被用于治疗全厚度烧伤伤口。然而，当发生大面积烧伤时，可供移植的供体位点较少，难以满足大规模皮肤移植的需求，增加了患者死亡的风险。本研究探讨了使用新近创建的低免疫原表皮细胞片来愈合皮肤创伤的可能性。采用Lentivirus转染技术生成过表达吲哚胺2,3-二氧化酶（IDO）的角质形成细胞（KCs）。通过Western blotting和定量聚合酶链反应测定IDO水平。通过CCK-8和Transwell实验评估IDO阳性角质形成细胞的功能。在角质形成细胞诱导培养基中培养KCs和成纤维细胞（FBs）以产生表皮细胞片。将BALB/c小鼠的全厚度皮肤切除物移植表皮细胞片。使用BALB/c裸鼠异种移植模型评估IDO阳性角质形成细胞的肿瘤发生性。在第12天通过CD3和CD31免疫荧光标记的创伤组织评估T淋巴细胞浸润和毛细血管发育。通过ELISA测定IL-1和TNF-α浓度。与对照组和阴性对照组相比，IDO阳性角质形成细胞显著增强了IDO mRNA和蛋白的表达水平，以及IDO阳性角质形成细胞的培养基中吡咯酮的含量。IDO组CD8+ T细胞的凋亡显著高于对照组和阴性对照组。然而，IDO阳性角质形成细胞的增殖和迁移能力与对照组和阴性对照组没有明显差异。无论是体外培养的低免疫原表皮细胞片还是体内移植实验都表明，IDO阳性表皮细胞片可以有效促进伤口愈合，且不具有肿瘤发生性。IDO阳性表皮细胞片可能通过降低伤口组织中炎症因子（TNF和IL-1）的表达水平、减少CD3+ T淋巴细胞的浸润和增加新生毛细血管来促进伤口愈合。本研究成功构建了低免疫原表皮细胞片，并证明低免疫原表皮细胞片能够加速伤口愈合。© 2023年日本再生医学学会。Elsevier B.V.制作和托管。

The physiological process of wound healing is dynamic, continuous, and intricate. Nowadays, full-thickness burn wounds are treated by autologous skin transplantation. Unfortunately, when substantial burns develop, there are fewer donor sites accessible, making it difficult to satisfy the requirement for large-scale skin transplants and increasing the risk of patient mortality. This study investigated the possibility of using a newly created hypoimmunogenic epidermal cell sheet to heal skin wounds.Transfection with lentivirus was used to generate Keratinocytes (KCs) that overexpress Indoleamine 2,3-Dioxygenase (IDO). Western blotting and quantitative polymerase chain reaction were used to measure IDO levels. To evaluate the function of IDO+ keratinocytes, CCK-8 and Transwell assays were performed. In cell sheet induction media, KCs and Fibroblasts (FBs) were cultured to yield epidermal cell sheets. The full-thickness skin excisions of BALB/c mice were transplanted with epidermal cell sheets. To assess the tumorigenicity of IDO+ keratinocytes, BALB/c nude mouse xenograft models were also used. CD3 and CD31 immunofluorescence labeling of wound tissue on day 12 to identify T lymphocyte infiltration and capillary development. ELISA measurement of IL-1 and TNF-α concentrations.IDO + keratinocytes dramatically enhanced the expression levels of IDO mRNA and protein, as well as the amount of kynurenine in the conditioned media of IDO+ keratinocytes, compared to the Control and NC groups. CD8+ T cell apoptosis was considerably greater in the IDO group than in the Control and NC groups. Nevertheless, the proliferation and migratory capabilities of IDO+ keratinocytes were not substantially different from those of the Control and NC groups. In vitro cultivation of the hypoimmunogenic epidermal cell sheet was effective. In vivo transplantation experiments demonstrated that IDO+ epidermal cell sheets can effectively promote wound healing without tumorigenicity, and IDO+ epidermal cell sheets may promote wound healing by decreasing the expression levels of inflammatory factors (TNF and IL-1) in wound tissue, decreasing CD3+ T lymphocytes, and increasing infiltration and new capillaries in wound tissue.In this study, we successfully constructed the hypoimmunogenic epidermal cell sheet and demonstrated that the hypoimmunogenic epidermal cell sheet could accelerate wound healing.© 2023 The Japanese Society for Regenerative Medicine. Production and hosting by Elsevier B.V.