Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA
Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA
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DOI：
10.1016/j.stemcr.2019.10.007
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发表时间：
2019-12-10
影响因子：
5.9
通讯作者：
Slukvin, Igor
中科院分区：
文献类型：
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作者：
Brok-Volchanskaya, Vera S.;Bennin, David A.;Slukvin, Igor
人类诱导多能干细胞（hiPSC）可以作为生物医学研究和输血治疗的嗜中性粒细胞的多功能和可扩展来源。在这里，我们描述了一种快速有效的无血清和无异种的中性粒细胞生成方案，该方案基于使用ETV 2修饰的mRNA对hiPSC进行直接的血液内皮编程。在GM-CSF、FGF 2和UM 171存在下培养ETV 2诱导的血内皮祖细胞导致连续产生大量的CD 34（+）CD 33（+）骨髓祖细胞，其可以每8-10天收获一次，培养长达30天。随后，在G-CSF和视黄酸激动剂Am 580的存在下，骨髓祖细胞分化成中性粒细胞。在这些条件下获得的中性粒细胞显示出典型的体细胞中性粒细胞形态，产生活性氧，形成中性粒细胞胞外陷阱，并具有吞噬和趋化活性。总的来说，该技术提供了在分化开始后14天就产生大量中性粒细胞的机会。
Human induced pluripotent stem cells (hiPSCs) can serve as a versatile and scalable source of neutrophils for biomedical research and transfusion therapies. Here we describe a rapid efficient serum- and xenogen-free protocol for neutrophil generation, which is based on direct hematoendothelial programming of hiPSCs using ETV2-modified mRNA. Culture of ETV2-induced hematoendothelial progenitors in the presence of GM-CSF, FGF2, and UM171 led to continuous production of generous amounts of CD34(+)CD33(+) myeloid progenitors which could be harvested every 8-10 days for up to 30 days of culture. Subsequently, myeloid progenitors were differentiated into neutrophils in the presence of G-CSF and the retinoic acid agonist Am580. Neutrophils obtained in these conditions displayed a typical somatic neutrophil morphology, produced reactive oxygen species, formed neutrophil extracellular traps and possessed phagocytic and chemotactic activities. Overall, this technology offers an opportunity to generate a significant number of neutrophils as soon as 14 days after initiation of differentiation.