A newly established megakaryoblastic/erythroid cell line that differentiates to red cells in the presence of erythropoietin and produces platelet‐like particles

Abstract

In August, 1992, we established a leukemic cell line (NS-Meg) from a patient in megakaryoblastic transformation of Philadelphia chromosome-positive chronic myeloid leukemia. The NS-Meg cells were positive for alpha-naphthyl acetate esterase and periodic acid-Schiff (PAS) staining and for surface CD4, CD7, CD13, CD34, CD41a, and glycophorin A antigens. Ultrastructurally, the cells had alpha-granules, demarcation membranes, and platelet peroxidase activity. The NS-Meg cells spontaneously produced platelet-like particles which contained alpha-granules, mitochondria and dense bodies, strongly suggesting platelet production. Erythropoietin (Epo), granulocyte/macrophage colony stimulating factor(GM-CSF), and interleukin 3 (IL-3) promoted the growth of NS-Meg cells. Phorbol-12-myristate-13-acetate increased the expression of both CD41a and CD61 antigens. Ten-day exposure to Epo induced mature erythroblasts and red cells. These benzidine-positive cells were positive for hemoglobin F staining. Untreated NS-Meg cells expressed mRNA for the Epo receptor (EpoR), for GATA-1, and for alpha 1, alpha 2 and gamma globin genes. These results indicate that NS-Meg cells undergo terminal differentiation of both megakaryocytic and erythroid lineages. This cell line should be a very useful tool for the investigation of both megakaryocytic and erythroid maturation.

Establishment and characterization of a new human megakaryoblastic cell line (SET-2) that spontaneously matures to megakaryocytes and produces platelet-like particles

“…Until now, all established megakaryoblastic and megakaryocytic cell lines have been derived from the megakaryoblastic crisis of patients with chronic myelogenous leukemia (CML) or with acute megakaryoblastic leukemia (FAB-M7 32 ). 9 Recently, a …”

“…Similar to MEG-A2 8 and NS-Meg, 9 SET-2 cells expressed the megakaryocytic lineage markers, CD4, 7, and HLA-DR antigens. Antigens detected in early hematopoietic progenitors, such as CD33 and 34 were also expressed in SET-2.…”

“…Platelet-like particles appear to be released by the dissociation of cytoplasms. Appearance of multinucleated giant cells in suspension culture has occasionally been detected in several cell lines, 2,[4][5][6]8,9,15 and CMK cells treated with PMA have been found to display morphologic characteristics of mature megakaryocytes with multilobular nuclei. 35 Among them, only three cell lines (MEG-01, M-07e and NS-Meg) have been shown to spontaneously produce platelet-like particles, suggesting megakaryocytic terminal differentiation.…”

“…37 Those from CMK11-5 possessed many of the same properties as normal platelets except that they were unable to aggregate. 36 NS-Meg, 9 a megakaryoblastic/erythroid cell line derived from a patient with CML in blast crisis, also spontaneously produces platelet-like particles. However, this cell line also did not exhibit a proliferative response to IL-6 or IL-11.…”

“…[1][2][3][4][5][6][7][8][9][10] Proliferation in some cytokine-dependent megakaryocytic leukemia cell lines is stimulated by TPO. [11][12][13] Expression of c-mpl is also observed in some megakaryocytic cell lines (eg M-07e and CMK).…”