The development of induced pluripotent stem cell-derived mesenchymal stem/stromal cells from normal human and RDEB epidermal keratinocytes
The development of induced pluripotent stem cell-derived mesenchymal stem/stromal cells from normal human and RDEB epidermal keratinocytes
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DOI：
10.1016/j.jdermsci.2018.06.004
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发表时间：
2018-09-01
影响因子：
4.6
通讯作者：
Shimizu, Hiroshi
中科院分区：
文献类型：
--
作者：
Nakayama, Chihiro;Fujita, Yasuyuki;Shimizu, Hiroshi
背景：大疱性表皮松解症(EB)是一组由真皮-表皮交界处(DEJ)结构分子编码基因突变引起的遗传性疾病。基于细胞的治疗，如异基因间充质干细胞/基质细胞(MSC)移植，最近已被探索用于治疗严重的EB类型，如隐性营养不良EB(RDEB)。然而，目前基于MSC的治疗存在障碍，如单一细胞来源的有限增殖和潜在的同种异体排斥反应限制细胞存活。目的：我们旨在从角质形成细胞来源的诱导多能干细胞(KC-IPSCs)中培养MSCs。方法：用激活素A、6-溴吲哚红-3‘-肟和骨形态发生蛋白4诱导RDEB患者和正常人的KC-IPSCs诱导中胚层谱系形成。对诱导后的细胞进行免疫组织化学分析、流式细胞仪分析和RNA芯片分析，并将其移植到创伤免疫缺陷小鼠的皮下和静脉内，观察其对创面愈合的影响。结果：经诱导后，KC-iPSC诱导的细胞与MSCs相容。此外，将KC-IPSC诱导的细胞注射到创伤免疫缺陷小鼠的皮下和静脉中，在上皮化区的DEJ处检测到人II型胶原。结论：我们成功地从正常人和RDEB患者的角质形成细胞中建立了IPSC来源的MSCs(KC-IPSC-MSCs)。KC-IPSC-MSCs有可能用于RDEB的治疗。(C)2018年日本皮肤病研究学会。爱思唯尔出版，版权所有。
Background: 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.Objectives: We aimed to develop MSCs from keratinocyte-derived induced pluripotent stem cells (iPSCs).Methods: 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.Results: 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.Conclusions: 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. (C) 2018 Japanese Society for Investigative Dermatology. Published by Elsevier B.V. All rights reserved.