Isolation and transcriptome analyses of human erythroid progenitors: BFU-E and CFU-E Journal Article
| Authors: | Li, J.; Hale, J.; Bhagia, P.; Xue, F.; Chen, L.; Jaffray, J.; Yan, H.; Lane, J.; Gallagher, P. G.; Mohandas, N.; Liu, J.; An, X. |
| Article Title: | Isolation and transcriptome analyses of human erythroid progenitors: BFU-E and CFU-E |
| Abstract: | Burst-forming unit-erythroid (BFU-E) and colony-forming unit-erythroid (CFU-E) cells are erythroid progenitors traditionally defined by colony assays.Wedeveloped a flowcytometrybased strategy for isolating human BFU-E and CFU-E cells based on the changes in expression of cell surface markers during in vitro erythroid cell culture. BFU-E and CFU-E are characterized by CD45+ GPA -IL-3R -CD34+CD36-CD71low and CD45+GPA- IL-3R-CD34-CD36+CD71high phenotypes, respectively. Colony assays validated phenotypic assignment giving rise to BFU-E and CFU-E colonies, both at a purity of ∼90%. The BFU-E colony forming ability of CD45+GPA-IL-3R-CD34+CD36-CD71low cells required stem cell factor and erythropoietin, while the CFU-E colony forming ability of CD45+GPA-IL-3R-CD34-CD36+CD71high cells required only erythropoietin. Bioinformatic analysis of the RNA-sequencing data revealed unique transcriptomes at each differentiation stage. Thesortingstrategywas validated inunculturedprimarycells isolated from bone marrow, cord blood, and peripheral blood, indicating that marker expression is not an artifact of in vitro cell culture, but represents an in vivo characteristic of erythroid progenitor populations. The ability to isolate highly pure human BFU-E and CFU-E progenitors will enable detailed cellular and molecular characterization of these distinct progenitor populations and define their contribution to disordered erythropoiesis in inherited and acquired hematologic disease.Our data provides an important resource for future studies of human erythropoiesis. |
| Keywords: | controlled study; protein expression; human cell; flow cytometry; phenotype; cd34 antigen; stem cell factor; gene expression; erythropoietin; erythroid precursor cell; erythropoiesis; in vitro study; cell population; cell surface marker; umbilical cord blood; bone marrow cell; transcriptome; cd45 antigen; differentiation; rna sequence; colony forming unit; rna analysis; rna isolation; cd36 antigen; burst forming unit e; cd71 antigen; human; article; colony forming unit e; interleukin 3 receptor; erythrocyte culture |
| Journal Title: | Blood |
| Volume: | 124 |
| Issue: | 24 |
| ISSN: | 0006-4971 |
| Publisher: | American Society of Hematology |
| Date Published: | 2014-12-04 |
| Start Page: | 3636 |
| End Page: | 3645 |
| Language: | English |
| DOI: | 10.1182/blood-2014-07-588806 |
| PROVIDER: | scopus |
| PMCID: | PMC4256913 |
| PUBMED: | 25339359 |
| DOI/URL: | |
| Notes: | Export Date: 2 January 2015 -- Source: Scopus |
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