U-Factor Co., Ltd, 1F, ESCALIER Rokubancho, 7-11, Rokubancho, Chiyoda, Tokyo, 102-0085, Japan.
Hitonowa Medical, K.PLAZA 2F, 1-7 Rokubancho, Chiyoda, Tokyo, 102-0085, Japan.
Sci Rep. 2024 Jul 3;14(1):15340. doi: 10.1038/s41598-024-66213-8.
Although stem cell-based regenerative medicine has been extensively studied, it remains difficult to reconstruct three dimensional tissues and organs in combination with vascular systems in vitro. One clinically successful therapy is transplantation of mesenchymal stem cells (MSC) into patients with graft versus host disease. However, transplanted cells are immediately damaged and destroyed because of innate immune reactions provoked by thrombogenic inflammation, and patients need to take immunosuppressive drugs for the immunological regulation of allogeneic cells. This reduces the benefits of stem cell transplantation. Therefore, alternative therapies are more realistic options for clinical use. In this study, we aimed to take advantage of the therapeutic efficacy of MSC and use multiple cytokines released from MSC, that is, stem cells from human exfoliated deciduous teeth (SHEDs). Here, we purified components from conditioned media of immortalized SHED (IM-SHED-CM) and evaluated the activities of intracellular dehydrogenase, cell migration, and antioxidative stress by studying the cells. The immortalization of SHED could make the stable supply of CM possible. We found that the fractionated component of 50-100 kD from IM-SHED-CM had higher efficacy than the original IM-SHED-CM in terms of intracellular dehydrogenase and cell migration in which intracellular signal transduction was activated via receptor tyrosine kinases, and the glutathione peroxidase and reductase system was highly active. Although antioxidative stress activities in the fractionated component of 50-100 kD had slightly lower than that of original IM-SHE-CM, the fraction still had the activity. Thus, the use of fractionated components of 50-100 kD from IM-SHED-CM could be an alternative choice for MSC transplantation because the purified components from CM could maintain the effect of cytokines from SHED.
尽管基于干细胞的再生医学已经得到了广泛的研究,但要在体外结合血管系统重建三维组织和器官仍然具有挑战性。一种临床成功的治疗方法是将间充质干细胞(MSC)移植到移植物抗宿主病患者体内。然而,由于血栓形成炎症引发的固有免疫反应,移植细胞会立即受损和破坏,并且患者需要服用免疫抑制剂来调节异基因细胞的免疫。这降低了干细胞移植的益处。因此,替代疗法是更现实的临床应用选择。在这项研究中,我们旨在利用 MSC 的治疗功效,并利用 MSC 释放的多种细胞因子,即来自人脱落乳牙的干细胞(SHED)。在这里,我们从永生化 SHED(IM-SHED-CM)的条件培养基中纯化成分,并通过研究细胞来评估细胞内脱氢酶、细胞迁移和抗氧化应激的活性。SHED 的永生化可以使 CM 的稳定供应成为可能。我们发现,来自 IM-SHED-CM 的 50-100kD 分馏成分在细胞内脱氢酶和细胞迁移方面比原始的 IM-SHED-CM 更有效,其中细胞内信号转导通过受体酪氨酸激酶激活,谷胱甘肽过氧化物酶和还原酶系统高度活跃。尽管 50-100kD 分馏成分的抗氧化应激活性略低于原始 IM-SHE-CM,但该分馏物仍具有活性。因此,使用来自 IM-SHED-CM 的 50-100kD 分馏成分可能是 MSC 移植的替代选择,因为 CM 中的纯化成分可以维持 SHED 细胞因子的作用。