Production of human hematopoietic survival and growth factor by a myeloid leukemia cell line (KPB-M15) and placenta as detected by a monoclonal antibody.
Fifty-five hematopoietic cell lines, including 19 T-, 16 B-, 5 pre-B-, 5 non-T non-B-, 1 erythroid, and 9 myeloid-monocytoid cells, were screened for production of human hematopoietic survival and stem cell growth factor (SCGF) by enzyme immunoassay using anti-SCGF monoclonal antibody. The KPB-M15 myeloid cell line constitutionally secreted a considerable quantity of SCGF, while other T- or myeloid-monocytoid cell lines did not secrete SCGF. Other biomaterials investigated were fetal calf, horse, and human serum; granulocyte-macrophage colony-stimulating factor and erythropoietin preparations; human placental conditioned medium; lectin (phytohemagglutinin, concanavalin A, and pokeweed mitogen); and mixed leukocyte reaction-stimulated leukocyte-conditioned medium. SCGF was detected only in human placental conditioned medium. SCGF produced by the KPB-M15 cells was a protein with a molecular weight of 20,000. The molecule, highly purified by immunoadsorbent affinity chromatography, retained SCGF activity in vitro, e.g., erythroid burst-promoting activity and granulocyte-macrophage-colony potentiation. With the availability of purified SCGF, it is now possible to study in detail the mechanisms regulating hematopoietic stem cells.
Hiraoka
A
A
Ohkubo
T
T
Fukuda
M
M
D000911:Antibodies, Monoclonal / Q000276:immunology*; D002460:Cell Line; D002846:Chromatography, Affinity; D003115:Colony-Stimulating Factors / Q000032:analysis*; D006133:Growth Substances / Q000032:analysis* / Q000302:isolation & purification / Q000494:pharmacology; D016298:Hematopoietic Cell Growth Factors; D006412:Hematopoietic Stem Cells / Q000187:drug effects*; D006801:Humans; D007124:Immunoenzyme Techniques; D007377:Interleukin-3 / Q000032:analysis; D007951:Leukemia, Myeloid / Q000378:metabolism*; D008970:Molecular Weight; D010920:Placenta / Q000032:analysis*
D000911:Antibodies, Monoclonal; D003115:Colony-Stimulating Factors; D006133:Growth Substances; D016298:Hematopoietic Cell Growth Factors; D007377:Interleukin-3