Bone marrow extracellular matrix induces HL-60 cells to produce an autonomous differentiation factor.
Conditioned medium from cultures of HL-60 myeloid leukemia cells grown on extracellular bone marrow matrix induces macrophage-like differentiation of fresh HL-60 cells. The active medium component is sensitive to protease treatment, indicating that it is a protein, but it is heat stable. Conditioned medium from HL-60 cells grown on protease-treated bone marrow matrix still contains the active component. Thus, it appears that the differentiation-inducing protein is produced by HL-60 cells and is not released from the bone marrow matrix. To identify this differentiation factor, RNA was isolated from HL-60 cells grown on bone marrow matrix and assayed by Northern analysis for expression of mRNA for human differentiation factor, tumor necrosis factor, and macrophage colony-stimulating factor, all inducers of monocyte/macrophage differentiation. Expression of differentiation factor, tumor necrosis factor, or macrophage colony-stimulating factor mRNA was not enhanced in HL-60 cells grown on matrix compared to cells grown on uncoated plastic flasks. Thus, the maturation factor does not appear to be differentiation factor, tumor necrosis factor, or macrophage colony-stimulating factor within the limits of detection of Northern analysis. Elution of the active conditioned medium fraction on a Sephacryl S-200 column revealed a molecular weight of approximately 40,000. The active protein eluted on a DEAE-cellulose ion-exchange column at an ionic strength of 0.3 M NaCl, indicating that it is fairly anionic. Thus, bone marrow matrix is able to induce HL-60 cells to produce a maturation-inducing 40 kilodalton protein.(ABSTRACT TRUNCATED AT 250 WORDS)
Mane
S
S
Department of Medicine, University of Minnesota Medical School, Minneapolis 55455.
Winkelmann
J C
JC
Luikart
S D
SD
D001685:Biological Factors / Q000096:biosynthesis*; D015152:Blotting, Northern; D001853:Bone Marrow / Q000502:physiology*; D002454:Cell Differentiation / Q000502:physiology; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D003470:Culture Media; D005109:Extracellular Matrix / Q000502:physiology*; D006497:Heparan Sulfate / Q000302:isolation & purification; D006801:Humans; D007951:Leukemia, Myeloid / Q000378:metabolism*; D011869:Radioligand Assay; D014407:Tumor Cells, Cultured
D001685:Biological Factors; D003470:Culture Media; D006497:Heparan Sulfate