Differentially expressed genes during in vitro differentiation of murine embryonic stem cells transduced with a human erythropoietin receptor cDNA

Z. B. Xia, Mushui Dai, C. Magoulas, H. E. Broxmeyer, L. Lu

Research output: Contribution to journalArticle

14 Citations (Scopus)

Abstract

Our previous study demonstrated that transduction of murine embryonic stem (ES) cells with a human erythropoietin (Epo) receptor (R) cDNA resulted in enhanced erythropoiesis in developing embryonic bodies (EBs). To address possible mechanisms of gene regulation, we compared gene expression between hEpoR cDNA-trasduced ES (ES-hEpoR) cells and parental ES cells during in vitro differentiation induced by withdrawal of leukemia inhibitory factor (LIF) and cultured in the absence of Epo using differential display reverse transcriptase-polymerase chain reaction (DDRT-PCR). A total of 48 differentially expressed cDNA fragments were found; 12 were sequenced and five were confirmed by Northern blot analysis to be up- or down-regulated in ES-hEpoR cells during differentiation compared to parental ES cells. In a GenBank search of the five putatively regulated cDNA fragments, two fragments shared high sequence homology to two known genes: the Surf-6 gene and the gene for calcyclin binding protein. Northern blot analysis demonstrated that 2.5-kb and 0.3-kb transcripts of the Surf-6 gene were expressed in undifferentiated ES-hEpoR and parental ES cells at a low level, but this expression was enhanced from day 2 to 14 of differentiation after withdrawal of LIF and culture in the presence of Epo. Furthermore, the enhanced expression of these two transcripts was also noticed in EML-C1 cells, a routine multipotential hematopoietic cell line that has erythroid differentiation potential in response to Epo. In summary, our results demonstrate that Surf-6 gene expression is regulated during differentiation of hematopoietic stem/progenitor cells in response to Epo, suggesting a possible role for Surf-6 gene in erythropoiesis.

Original languageEnglish (US)
Pages (from-to)651-658
Number of pages8
JournalJournal of Hematotherapy and Stem Cell Research
Volume9
Issue number5
DOIs
StatePublished - 2000
Externally publishedYes

Fingerprint

Erythropoietin Receptors
Embryonic Stem Cells
Complementary DNA
Erythropoietin
Genes
Leukemia Inhibitory Factor
Erythropoiesis
Hematopoietic Stem Cells
Northern Blotting
Nucleic Acid Databases
Gene Expression Regulation
Sequence Homology
Reverse Transcriptase Polymerase Chain Reaction
In Vitro Techniques
Cell Differentiation
Carrier Proteins
Gene Expression
Cell Line

ASJC Scopus subject areas

  • Hematology
  • Immunology

Cite this

Differentially expressed genes during in vitro differentiation of murine embryonic stem cells transduced with a human erythropoietin receptor cDNA. / Xia, Z. B.; Dai, Mushui; Magoulas, C.; Broxmeyer, H. E.; Lu, L.

In: Journal of Hematotherapy and Stem Cell Research, Vol. 9, No. 5, 2000, p. 651-658.

Research output: Contribution to journalArticle

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AU - Lu, L.

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