Department of Dermatology, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
Department of Dermatology, Hokkaido University Graduate School of Medicine, Sapporo, Japan.
J Dermatol Sci. 2018 Sep;91(3):301-310. doi: 10.1016/j.jdermsci.2018.06.004. Epub 2018 Jun 18.
Epidermolysis bullosa (EB) is a group of hereditary disorders caused by mutations in the genes encoding structural molecules of the dermal-epidermal junction (DEJ). Cell-based therapies such as allogeneic mesenchymal stem/stromal cell (MSC) transplantation have recently been explored for severe EB types, such as recessive dystrophic EB (RDEB). However, hurdles exist in current MSC-based therapies, such as limited proliferation from a single cell source and limited cell survival due to potential allogenic rejection.
We aimed to develop MSCs from keratinocyte-derived induced pluripotent stem cells (iPSCs).
Keratinocyte-derived iPSCs (KC-iPSCs) of a healthy human and an RDEB patient were cultured with activin A, 6-bromoindirubin-3'-oxime and bone morphogenetic protein 4 to induce mesodermal lineage formation. These induced cells were subjected to immunohistochemical analysis, flow cytometric analysis and RNA microarray analysis in vitro, and were injected subcutaneously and intravenously to wounded immunodeficient mice to assess their wound-healing efficacy.
After their induction, KC-iPSC-induced cells were found to be compatible with MSCs. Furthermore, with the subcutaneous and intravenous injection of the KC-iPSC-induced cells into wounded immunodeficient mice, human type VII collagen was detected at the DEJ of epithelized areas.
We successfully established iPSC-derived MSCs from keratinocytes (KC-iPSC-MSCs) of a normal human and an RDEB patient. KC-iPSC-MSCs may have potential in therapies for RDEB.
大疱性表皮松解症（EB）是一组遗传性疾病，由编码表皮-真皮连接（DEJ）结构分子的基因突变引起。最近，人们探索了基于细胞的疗法，如同种异体间充质干细胞（MSC）移植，用于治疗严重的 EB 类型，如隐性营养不良性 EB（RDEB）。然而，目前基于 MSC 的治疗方法存在一些障碍，例如单个细胞来源的增殖有限，以及由于潜在的同种异体排斥而导致的细胞存活率有限。
我们旨在从角蛋白细胞衍生的诱导多能干细胞（iPSC）中开发 MSC。
培养健康人和 RDEB 患者的角蛋白细胞衍生的 iPSC（KC-iPSC），用激活素 A、6-溴靛玉红-3'-肟和骨形态发生蛋白 4 诱导中胚层谱系形成。这些诱导细胞进行体外免疫组织化学分析、流式细胞术分析和 RNA 微阵列分析，并皮下和静脉注射到受伤的免疫缺陷小鼠中，以评估其伤口愈合效果。
诱导后，发现 KC-iPSC 诱导的细胞与 MSC 相容。此外，将 KC-iPSC 诱导的细胞皮下和静脉注射到受伤的免疫缺陷小鼠中，在表皮化区域的 DEJ 处检测到人类 VII 型胶原蛋白。
我们成功地从正常人的角蛋白细胞和 RDEB 患者的角蛋白细胞中建立了 iPSC 衍生的 MSC（KC-iPSC-MSCs）。KC-iPSC-MSCs 可能在 RDEB 的治疗中有潜力。
