Differentiation signalobody: Demonstration of antigen-dependent osteoclast differentiation from a progenitor cell line
Differentiation signalobody: Demonstration of antigen-dependent osteoclast differentiation from a progenitor cell line
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DOI：
10.1016/j.jbiosc.2016.02.010
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发表时间：
2016-09-01
影响因子：
2.8
通讯作者：
Nagamune, Teruyuki
中科院分区：
文献类型：
--
作者：
Nakabayashi, Hideto;Aoyama, Saeko;Nagamune, Teruyuki
一种“无细胞质”的体外分化方法将有希望用于再生医学的细胞的成本效益大规模生产。在这项研究中，我们开发了一种分化信号体S-RANK，其中核因子κ B受体激活剂（RANK）的胞外结构域被替换为单链可变片段（scFv），以获得响应廉价抗原的信号传导。用S-RANK慢病毒转导鼠巨噬细胞系RAW 264（已知其通过RANK配体（RANKL）分化为破骨细胞）。当所得细胞与特异性抗原一起培养时，细胞分化成多核抗酒石酸盐酸性磷酸酶阳性破骨细胞。分化效率与RANKL诱导的分化效率几乎相当。此外，信号传导分析表明，野生型RANK下游的主要信号传导途径核因子κ B和丝裂原活化蛋白激酶信号传导途径也被S-RANK激活。这些结果表明，S-RANK充分模拟了野生型RANK的信号转导。分化信号体可以通过使用适当的信号传导结构域用于控制其他细胞类型的分化。(C)2016年，日本生物技术学会。All rights reserved.
A "cytoldne-less" in vitro differentiation method would be promising for cost-effective mass production of cells used for regenerative medicine. In this study, we developed a differentiation signalobody S-RANK, in which the extracellular domain of receptor activator of nuclear factor kappa-B (RANK) is replaced with a single-chain variable fragment (scFv) to attain signaling in response to an inexpensive antigen. A murine macrophage cell line RAW264, which is known to differentiate into an osteoclast by RANK ligand (RANKL), was lentivirally transduced with S-RANK. When the resultant cells were cultured with a specific antigen, the cells differentiated into multinucleated tartrate-resistant acid phosphatase-positive osteoclasts. The differentiation efficiency was almost comparable to those induced by RANKL. In addition, the signaling analysis demonstrated that nuclear factor kappa-B and mitogen-activated protein kinase signaling pathways, which are the major signaling pathways downstream of wild-type RANK, were also activated by S-RANK. These results demonstrate that S-RANK sufficiently mimics signal transduction of wild-type RANK. Differentiation signalobodies may be applied for controlling differentiation of other cell types by using appropriate signaling domains. (C) 2016, The Society for Biotechnology, Japan. All rights reserved.