An erythroid and megakaryocytic common precursor cell line (B1647) expressing both c-mpl and erythropoietin receptor (Epo-R) proliferates and modifies globin chain synthesis in response to megakaryocyte growth and development factor (MGDF) but not to erythropoietin (Epo).
A human megakaryocyte cell line (B1647) has been established from bone marrow cells obtained from a patient with acute myelogenous leukaemia (FAB M2). The cells were CD34-, CD33+, HLA-DR+, CD38+, and expressed the immunophenotypic markers of the megakaryocyte lineage (CD41 and von Willebrand factor). Moreover the cells expressed the c-mpl (thrombopoietin receptor) mRNA and protein. On the other hand, the B1647 cells also possessed erythroid lineage characteristics: the vast majority of cells were glycophorin positive, and about 10% of unstimulated cells stained with an anti-globin gamma chain MoAb. In addition, S1 protection analysis demonstrated expression of beta-globin mRNA, and Epo receptor (Epo-R) protein was detected by cytofluorimetric assay. Several growth factors, when tested alone or in combination, failed to influence the B1647 cell growth. A significant increase of cell proliferation was observed only after the addition, in serum-free culture, of recombinant human megakaryocyte growth development factor (MGDF), a recombinant c-mpl ligand encompassing the receptor-binding domain and identical to thrombopoietin (TPO), at concentrations ranging from 0.01 to 1 ng/ml. Interestingly, MGDF failed to induce megakaryocytic differentiation of the B1647 cells, but significantly increased the synthesis of the globin gamma-chain. B1647 cells could be a useful model for studying the biological effect of TPO on common megakaryocyte and erythroid progenitors.
Bonsi
L
L
Institute of Histology and Embryology, University of Bologna, Italy.
Grossi
A
A
Strippoli
P
P
Tumietto
F
F
Tonelli
R
R
Vannucchi
A M
AM
Ronchi
A
A
Ottolenghi
S
S
Visconti
G
G
Avanzi
G C
GC
Pegoraro
L
L
Bagnara
G P
GP
D002455:Cell Division; D002460:Cell Line; D016207:Cytokines / Q000494:pharmacology; D015672:Erythroid Precursor Cells / Q000166:cytology / Q000378:metabolism*; D004921:Erythropoietin / Q000494:pharmacology*; D005914:Globins / Q000096:biosynthesis*; D006801:Humans; D008533:Megakaryocytes / Q000166:cytology / Q000378:metabolism*; D010641:Phenotype; D012333:RNA, Messenger / Q000378:metabolism; D017467:Receptors, Erythropoietin / Q000378:metabolism*; D012701:Serotonin / Q000494:pharmacology; D013926:Thrombopoietin / Q000494:pharmacology*
D016207:Cytokines; D012333:RNA, Messenger; D017467:Receptors, Erythropoietin; D004921:Erythropoietin; D012701:Serotonin; D005914:Globins; D013926:Thrombopoietin