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Published on: January 6, 2018
Identification of differentiation-inducing activity produced by human bone marrow stromal cell line LP101
Masaki Hiramoto1, Yutaka Kawakami, Ryusuke Nabeshima
1Department of Anatomy, Nihon University School of Medicine, Oyaguchikamicho, Itabashi-ku, Tokyo 173-8610, Japan.
Insights
Human bone marrow stromal cells (LP101) induce leukemia cell differentiation via cytokines like TNF-alpha, IL-6, and GM-CSF. Low molecular weight factors also contribute to this granulocyte differentiation process.
Area of Science:
- Hematology
- Cell Biology
- Immunology
Background:
- Human promyelocytic leukemia HL-60 cells can differentiate into mature granulocytes when co-cultured with human bone marrow stromal LP101 cells.
- Understanding the specific factors responsible for this differentiation is crucial for potential therapeutic applications.
Purpose of the Study:
- To identify the specific factors produced by LP101 cells that induce HL-60 cell differentiation into mature granulocytes.
- To elucidate the roles of various cytokines and other factors in this differentiation process.
Main Methods:
- HL-60 cells were cultured with conditioned medium (CM) from LP101 cells.
- Cytokine expression in CM was analyzed, and neutralizing antibodies were used.
- Recombinant cytokines were tested for their differentiation-inducing effects.
- CM was fractionated by ultrafiltration into low (LMW) and high (HMW) molecular weight fractions.
Main Results:
- CM from LP101 cells increased CD11b expression on HL-60 cells, indicating differentiation.
- LP101 cells produced TNF-alpha, GM-CSF, and IL-6.
- Neutralizing antibodies against these cytokines partially inhibited CM-induced differentiation.
- Recombinant TNF-alpha induced differentiation, enhanced by GM-CSF and IL-6.
- LMW fraction of CM synergistically enhanced TNF-alpha's differentiation-inducing activity.
Conclusions:
- LP101 cells induce HL-60 cell differentiation through the production of TNF-alpha, IL-6, and GM-CSF.
- Unknown low molecular weight factors produced by LP101 cells also play a synergistic role in inducing HL-60 cell differentiation.
Abstract:
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.
