Identification of differentiation-inducing activity produced by human bone marrow stromal cell line LP101.
We have previously reported that human promyelocytic leukemia HL-60 cells can be induced to differentiate into mature granulocytes when HL-60 co-cultivated with human bone marrow stromal LP101 cells. In the present study, we investigated which factors produced by LP101 cells induce HL-60 cells to differentiate into mature granulocytes. The expression of the cell surface antigen CD11b on HL-60 cells was increased after a 72-h culture with the conditioned medium (CM) obtained from LP101 cells. LP101 cells were observed to produce various cytokines, including TNF-alpha, GM-CSF and IL-6. The neutralizing antibodies against these cytokines partially suppressed the CM-induced differentiation of HL-60 cells. Recombinant TNF-alpha induced the differentiation of HL-60 cells, and GM-CSF and IL-6 additionally enhanced the effect of TNF-alpha. When the CM was divided into a low molecular weight (LMW) fraction and a high molecular weight (HMW) fraction by ultrafiltration, the LMW fraction synergistically enhanced the differentiation inducible activity of TNF-alpha. These results demonstrate that LP101 cells induce the differentiation of HL-60 cells by producing various cytokines including TNF-alpha, IL-6, and GM-CSF, and that unknown low molecular weight factors also participate.
Hiramoto
Masaki
M
Department of Anatomy, Nihon University School of Medicine, Oyaguchikamicho, Itabashi-ku, Tokyo 173-8610, Japan.
Kawakami
Yutaka
Y
Nabeshima
Ryusuke
R
Shima
Daisuke
D
Handa
Hiroshi
H
Aizawa
Shin
S
D000906:Antibodies / Q000494:pharmacology; D001854:Bone Marrow Cells / Q000166:cytology*; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D017077:Culture Media, Conditioned; D016207:Cytokines / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000494:pharmacology; D017931:DNA Primers; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000494:pharmacology; D018922:HL-60 Cells; D006801:Humans; D015850:Interleukin-6 / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017154:Stromal Cells / Q000166:cytology*; D014409:Tumor Necrosis Factor-alpha / Q000494:pharmacology
D000906:Antibodies; D017077:Culture Media, Conditioned; D016207:Cytokines; D017931:DNA Primers; D015850:Interleukin-6; D012333:RNA, Messenger; D014409:Tumor Necrosis Factor-alpha; D016178:Granulocyte-Macrophage Colony-Stimulating Factor