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STAR PROTOCOLS
Volume 1, Issue 2, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.xpro.2020.100075
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Funding
- UW2020 Wisconsin Alumni Research Foundation (WARF
- United States) program
- National Institute of Health, United States [R01AI134749-01, R01HL142665, P51 OD011106]
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This protocol describes a rapid and efficient feeder-, serum-, and xeno-free method for neutrophil generation from hiPSCs using ETV2 modified mRNA (mmRNA), which directs hematoendothelial programming of hiPSCs. Hematoen-dothelial progenitors were cultured with GM-CSF, FGF-2, and UM171 to expand myelomonocytic progenitors, followed by treatment with G-CSF and retinoic acid agonist Am580 to induce neutrophil maturation. This protocol is suitable for generating functional neutrophils from iPSCs to interrogate the role of genes in a neutrophil development and function.For complete details on the use and execution of this protocol, please refer to Brok-Volchanskaya et al. (2019).
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