RRC ID 4341
Author Saeki K, Saeki K, Nakahara M, Matsuyama S, Nakamura N, Yogiashi Y, Yoneda A, Koyanagi M, Kondo Y, Yuo A.
Title A feeder-free and efficient production of functional neutrophils from human embryonic stem cells.
Journal Stem Cells
Abstract A novel, feeder-free hematopoietic differentiation protocol was established for highly efficient production of neutrophils from human embryonic stem cells (hESCs). For the induction of differentiation, spheres were generated in the presence of serum and cytokine cocktail and subjected to attachment culture on gelatin-coated plates. After approximately 2 weeks, a sac-like structure filled with abundant round cells emerged at the center of flattened spheres. After cutting off this sac-like structure, round cells actively proliferated, either floating in the supernatant or associated weakly with the adherent cells. Almost all of these round cells were CD45-positive hematopoietic cells with myeloid phagocytic markers (CD33 and CD11b), and approximately 30%-50% of the round cells were mature neutrophils, as judged from morphology, cytochemical characteristics (myeloperoxidase and neutrophil alkaline phosphatase), and neutrophil-specific cell surface markers (CD66b, CD16b, and GPI-80). In addition, hESC-derived neutrophils had chemotactic capacity in response to the bacterial chemotactic peptide formyl-methionyl-leucyl-phenylalanine and neutrophil-specific chemokine interleukin (IL)-8. Using "semipurified" neutrophils migrated to IL-8, both phagocytic and respiratory burst activities were demonstrated. Finally, it was shown that hESC-derived neutrophils had chemotactic activity in vivo in a murine air-pouch inflammatory model. The present results indicate successful induction of functional mature neutrophils from hESCs via highly efficient feeder-free differentiation culture system of human hematopoietic cells.
Volume 27(1)
Pages 59-67
Published 2009-1
DOI 10.1634/stemcells.2007-0980
PII 2007-0980
PMID 18845766
MeSH Animals Cell Culture Techniques / methods* Cell Differentiation / drug effects Cell Line Cell Shape / drug effects Chemotaxis / drug effects Colony-Forming Units Assay Embryonic Stem Cells / cytology* Embryonic Stem Cells / drug effects Embryonic Stem Cells / metabolism Flow Cytometry Hematopoiesis / drug effects Humans Inflammation / pathology Interleukin-8 / pharmacology Karyotyping Mice Mice, SCID N-Formylmethionine Leucyl-Phenylalanine / pharmacology Neutrophils / cytology* Neutrophils / drug effects Neutrophils / metabolism Phagocytosis / drug effects Respiratory Burst / drug effects Transcription, Genetic / drug effects
IF 5.587
Times Cited 29
Human embryonic stem cells KhES-1 KhES-2 KhES-3