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Screening method for colony-stimulating factor inducers using a human bone marrow stromal cell line, KM-102
T Kohama1, H Miyaoka, A Torikata
1Bioscience Research Laboratories, Sankyo Co., Ltd., Tokyo, Japan.
The Journal of Antibiotics
|October 1, 1993
Summary
A novel assay effectively screens for colony-stimulating factor (CSF) inducers. This method identified two actinomycete strains producing novel CSF-inducing compounds from microbial screening.
Area of Science:
- Biotechnology
- Cell Biology
- Immunology
Background:
- Colony-stimulating factors (CSFs) are crucial for hematopoiesis.
- Existing methods for identifying CSF inducers can be inefficient.
- A robust screening system is needed to discover novel CSF-modulating agents.
Purpose of the Study:
- To develop and validate a new screening assay for identifying inducers of colony-stimulating factors (CSFs).
- To utilize this assay to screen microbial strains for novel CSF production.
- To characterize the identified CSF-inducing substances.
Main Methods:
- Established a novel assay combining CSF-dependent cell lines with a CSF-producing human bone marrow stromal cell line (KM-102).
- Utilized Interleukin-1 (IL-1) as a positive control for granulocyte CSF (G-CSF) and granulocyte-macrophage CSF (GM-CSF) production.
- Screened 1,600 microbial strains using the developed assay system.
Main Results:
- The assay successfully detected CSF induction within 24 hours.
- Activators of protein kinase C (PKC), protein phosphatase inhibitors, and lipopolysaccharide (LPS) tested positive.
- Two actinomycete strains were identified as producers of active substances, including three novel compounds with CSF-inducing properties.
Conclusions:
- The developed screening method is effective for identifying CSF inducers.
- Novel CSF-inducing compounds were discovered from microbial sources.
- These novel compounds represent a new class of CSF inducers, distinct from teleocidin-like substances.