Effective and Rapid Generation of Functional Neutrophils from Induced Pluripotent Stem Cells Using ETV2-Modified mRNA

Autor: David A. Bennin, Vera Brok-Volchanskaya, Anna Huttenlocher, Lucas C. Klemm, Kran Suknuntha, Igor I. Slukvin
Jazyk: angličtina
Rok vydání: 2019
Předmět:
Resource
0301 basic medicine
Myeloid
Neutrophils
Somatic cell
Induced Pluripotent Stem Cells
CD33
Retinoic acid
Biology
Extracellular Traps
Biochemistry
Immunophenotyping
03 medical and health sciences
chemistry.chemical_compound
0302 clinical medicine
modified mRNA
Genetics
medicine
Humans
RNA
Messenger
Progenitor cell
hemogenic endothelium
Induced pluripotent stem cell
lcsh:QH301-705.5
Cells
Cultured
Myeloid Progenitor Cells
lcsh:R5-920
ETV2
Gene Expression Regulation
Developmental
Cell Differentiation
Chemotaxis
Cell Biology
Neutrophil extracellular traps
myeloid progenitors
Hematopoiesis
Cell biology
030104 developmental biology
medicine.anatomical_structure
chemistry
lcsh:Biology (General)
Leukopoiesis
human iPSCs
hematopoietic differentiation
lcsh:Medicine (General)
Biomarkers
030217 neurology & neurosurgery
Transcription Factors
Developmental Biology
Zdroj: Stem Cell Reports, Vol 13, Iss 6, Pp 1099-1110 (2019)
Stem Cell Reports
ISSN: 2213-6711
Popis: Summary 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.
Graphical Abstract
Highlights • ETV2 mmRNA directly programs hPSCs into hemogenic endothelium (HE) • ETV2-induced HE possesses robust myeloid potential • ETV2 mmRNA rapid neutrophil differentiation protocol in defined conditions is provided • ETV2 mmRNA-induced neutrophils are functionally similar to in-vivo-derived cells
In this article, Slukvin and colleagues describes a protocol for a rapid efficient feeder-, serum-, and xenogen-free protocol for neutrophil generation from hiPSCs, which is based on direct hematoendothelial programming of hiPSCs using ETV2-modified mRNA.
Databáze: OpenAIRE
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