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Publication : Generation of eosinophils from cryopreserved murine bone marrow cells.

First Author  Schollaert KL Year  2014
Journal  PLoS One Volume  9
Issue  12 Pages  e116141
PubMed ID  25551463 Mgi Jnum  J:225660
Mgi Id  MGI:5693998 Doi  10.1371/journal.pone.0116141
Citation  Schollaert KL, et al. (2014) Generation of eosinophils from cryopreserved murine bone marrow cells. PLoS One 9(12):e116141
abstractText  Eosinophils are produced in the bone marrow from CD34+ eosinophil lineage-committed progenitors, whose levels in the bone marrow are elevated in a variety of human diseases. These findings suggest that increased eosinophil lineage-committed progenitor production is an important process in disease-associated eosinophilia. The pathways central to the biology of the eosinophil lineage-committed progenitor remain largely unknown. Thus, developing new methods to investigate the regulators of eosinophil lineage-committed progenitor differentiation is needed to identify potential therapeutic targets to specifically inhibit eosinophil production. We tested cytokine regimens to optimize liquid cultures for the study of eosinophil lineage-committed progenitor and eosinophil precursor differentiation into mature eosinophils. Stem cell factor (but not fms-related tyrosine kinase 3 ligand) was required for optimal yield of eosinophils. Furthermore, we evaluated the effects of cell preservation and scale on the culture, successfully culturing functional eosinophils from fresh and frozen murine bone marrow cells and in a standard-sized and 96-well culture format. In summary, we have developed an adaptable culture system that yields functionally competent eosinophils from murine low-density bone marrow cells and whose cytokine regime includes expansion of progenitors with stem cell factor alone with subsequent differentiation with interleukin 5.
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