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A Flow-cytometric approach to Eythropoiesis

We pioneered the use of cell-surface markers for the staging of murine erythroid progenitors within hematopoietic tissue (Koulnis et al., 2011; Liu et al., 2006; Pop et al., 2010; Socolovsky et al., 2001; Zhang et al., 2003). We make use of the cell surface markers CD71 and Ter119 to define progenitor subsets and simultaneously probe their cell viability, cell cycle status or gene expression profiles, using multiparameter flow cytometry, multispectral imaging, or sorting and subsequent molecular analysis. This approach allows us to study minimally-perturbed primary erythroid progenitors directly within the context of their physiological niche in vivo.

Flow Cytometry

Identification of erythroid progenitors and precursors within mouse fetal liver or other hematopoietic tissue using cell surface markers CD71 and Ter119. The cells can be isolated and analyzed by flow cytometry.

Flow Cytometry

Fetal liver erythroid cells are subdivided into subsets S0 to S5, containing increasingly-mature erythroblasts, based on their CD71/Ter119 expression.


References:

Koulnis, M., Pop, R., Porpiglia, E., Shearstone, J.R., Hidalgo, D., and Socolovsky, M. (2011). Identification and Analysis of Mouse Erythroid Progenitors using the CD71/TER119 Flow-cytometric Assay. J Vis Exp.

Liu, Y., Pop, R., Sadegh, C., Brugnara, C., Haase, V.H., and Socolovsky, M. (2006). Suppression of Fas-FasL coexpression by erythropoietin mediates erythroblast expansion during the erythropoietic stress response in vivo. Blood 108, 123-133.

Pop, R., Shearstone, J.R., Shen, Q., Liu, Y., Hallstrom, K., Koulnis, M., Gribnau, J., and Socolovsky, M. (2010). A key commitment step in erythropoiesis is synchronized with the cell cycle clock through mutual inhibition between PU.1 and S-phase progression. PLoS Biol 8.

Socolovsky, M., Nam, H., Fleming, M.D., Haase, V.H., Brugnara, C., and Lodish, H.F. (2001). Ineffective erythropoiesis in Stat5a(-/-)5b(-/-) mice due to decreased survival of early erythroblasts. Blood 98, 3261-3273.

Zhang, J., Socolovsky, M., Gross, A.W., and Lodish, H.F. (2003). Role of Ras signaling in erythroid differentiation of mouse fetal liver cells: functional analysis by a flow cytometry-based novel culture system. Blood 102, 3938-3946.