Ex vivo Generation of Genetically Modified Macrophages from Human Induced Pluripotent Stem Cells
Ex vivo Generation of Genetically Modified Macrophages from Human Induced Pluripotent Stem Cells
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DOI：
10.1159/000477129
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发表时间：
2017-01-01
影响因子：
2.2
通讯作者：
Lachmann, Nico
中科院分区：
文献类型：
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作者：
Ackermann, Mania;Kuhn, Alexandra;Lachmann, Nico
背景资料：多能干细胞，包括诱导多能干细胞（iPSC），具有向所有三个胚层分化的能力，并且已被强调为再生医学领域的有吸引力的细胞来源。因此，治疗性转基因在iPSC及其衍生的造血谱系后代中的稳定表达可以为创新的细胞替代疗法奠定基础。研究方法：我们已经利用通过慢病毒载体技术或报告基因的靶向整合遗传修饰的人iPSC系来评估造血特化和向巨噬细胞分化期间的转基因表达。结果如下：使用配备有普遍存在的染色质开放元件（CBX 3-UCOE）的慢病毒载体以及锌指核酸酶介导的表达盒靶向人腺相关病毒整合位点1（AAVS 1）安全港导致iPSC中稳定的转基因表达。当iPSC沿髓系途径沿着为巨噬细胞时，两种策略在造血特化过程中产生持续的转基因表达，包括成熟的CD 14+和CD 11b+巨噬细胞。结论：人iPSC技术与慢病毒载体技术或基于设计者核酸酶的基因组编辑的组合允许产生具有稳定转基因表达的转基因iPSC衍生的巨噬细胞，其可用于新型细胞和基因替代疗法。(C)2017 S. Karger GmbH，弗赖堡
Background: Pluripotent stem cells, including induced pluripotent stem cells (iPSCs), have the capacity to differentiate towards all three germ layers and have been highlighted as an attractive cell source for the field of regenerative medicine. Thus, stable expression of therapeutic transgenes in iPSCs, as well as thereof derived progeny of hematopoietic lineage, may lay the foundation for innovative cell replacement therapies. Methods: We have utilized human iPSC lines genetically modified by lentiviral vector technology or targeted integration of reporter genes to evaluate transgene expression during hematopoietic specification and differentiation towards macrophages. Results: Use of lentiviral vectors equipped with an ubiquitous chromatin opening element (CBX3-UCOE) as well as zinc finger nuclease-mediated targeting of an expression cassette into the human adeno-associated virus integration site 1 (AAVS1) safe harbor resulted in stable transgene expression in iPSCs. When iPSCs were differentiated along the myeloid pathway into macrophages, both strategies yielded sustained transgene expression during the hematopoietic specification process including mature CD14+ and CD11b+ macrophages. Conclusion: Combination of human iPSC technology with either lentiviral vector technology or designer nuclease-based genome editing allows for the generation of transgenic iPSC-derived macrophages with stable transgene expression which may be useful for novel cell and gene replacement therapies. (C) 2017 S. Karger GmbH, Freiburg