Two-Step Differentiation of Mast Cells from Induced Pluripotent Stem Cells
Two-Step Differentiation of Mast Cells from Induced Pluripotent Stem Cells
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DOI：
10.1089/scd.2012.0339
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发表时间：
2013-03-01
影响因子：
4
通讯作者：
Kawabata, Kenji
中科院分区：
文献类型：
--
作者：
Yamaguchi, Tomoko;Tashiro, Katsuhisa;Kawabata, Kenji
肥大细胞在变态反应性疾病的发病机制中起重要作用。它们通常分为2个表型不同的群体：结缔组织型肥大细胞（CTMCs）和粘膜型肥大细胞（MMCs）。从组织中获得的肥大细胞数量有限，这使得研究肥大细胞的功能变得困难。在这里，我们报告的CTMC样肥大细胞来源于小鼠诱导多能干细胞（iPS）的生成和表征。通过0 P9共培养方法或胚状体形成方法产生iPS细胞衍生的肥大细胞（iPSMC）。与骨髓来源的肥大细胞（BMMC）相比，通过两种方法分化的iPSMC中番红O阳性细胞的数量、CD 81蛋白和组氨酸脱羧酶mRNA的表达水平以及蛋白酶活性升高。电子显微镜分析显示，iPSMC比BMMC含有更多的颗粒。在用阳离子促分泌剂或万古霉素刺激后，诱导iPSMC脱粒。此外，iPSMC具有在体外和体内对IgE/抗原复合物的刺激作出响应的能力。此外，当在OP 9细胞上产生的iPSMC与Swiss 3 T3成纤维细胞共培养时，作为成熟指标的蛋白酶活性更高，表明成熟肥大细胞从iPS细胞分化。iPSMC可用作CTMC的体外模型以研究其功能。
Mast cells play important roles in the pathogenesis of allergic diseases. They are generally classified into 2 phenotypically distinct populations: connective tissue-type mast cells (CTMCs) and mucosal-type mast cells (MMCs). The number of mast cells that can be obtained from tissues is limited, making it difficult to study the function of mast cells. Here, we report the generation and characterization of CTMC-like mast cells derived from mouse induced pluripotent stem (iPS) cells. iPS cell-derived mast cells (iPSMCs) were generated by the OP9 coculture method or embryoid body formation method. The number of Safranin O-positive cells, expression levels of CD81 protein and histidine decarboxylase mRNA, and protease activities were elevated in the iPSMCs differentiated by both methods as compared with those in bone marrow-derived mast cells (BMMCs). Electron microscopic analysis revealed that iPSMCs contained more granules than BMMCs. Degranulation was induced in iPSMCs after stimulation with cationic secretagogues or vancomycin. In addition, iPSMCs had the ability to respond to stimulation with the IgE/antigen complex in vitro and in vivo. Moreover, when iPSMCs generated on OP9 cells were cocultured with Swiss 3T3 fibroblasts, protease activities as maturation index were more elevated, demonstrating that mature mast cells were differentiated from iPS cells. iPSMCs can be used as an in vitro model of CTMCs to investigate their functions.