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CSF-responsive bone marrow cells in liquid culture: characterization and screening applications
International Journal of Cell Cloning
|January 1, 1986
Summary
Liquid cultures effectively screen for colony-stimulating factor (CSF) and assess marrow toxicity. This method closely mimics conventional assays, offering reliable insights into myeloid cell production and differentiation.
Area of Science:
- Hematology
- Cell Biology
- Immunology
Background:
- Previous studies established a correlation between colony-stimulating factor (CSF) activity and 3H-thymidine ([3H]TdR) uptake in human marrow cells.
- The need for efficient screening methods for CSF and immunotoxin-induced marrow toxicity prompted the investigation of liquid culture assays.
Purpose of the Study:
- To compare the efficacy of liquid cell cultures with conventional granulocyte/macrophage colony-forming unit (CFU-gm) assays for screening CSF and assessing marrow toxicity.
- To evaluate the correlation between [3H]TdR uptake and myeloid cell production/differentiation in liquid cultures.
Main Methods:
- Comparison of CSF dose-response curves in liquid cultures versus CFU-gm colony assays.
- Assessment of [3H]TdR uptake in nonadherent marrow cells and its correlation with colony and cluster growth.
- Analysis of myeloid cell differentiation, surface antigen expression (Ia-like antigens, transferrin receptors), and DNA content in liquid cultures.
- Evaluation of ricin and anti-Ia cytotoxicity in both liquid and CFU-gm cultures.
Main Results:
- CSF dose-response relationships were nearly identical between liquid and colony-forming assays.
- [3H]TdR uptake in liquid cultures showed a strong correlation with CFU-gm colony growth and was dose-dependent on CSF.
- Liquid cultures accurately reflected myeloid cell differentiation and allowed for quantitative assessment of cell production, similar to colony cultures.
- Cytotoxicity of immunotoxins (ricin, anti-Ia) was comparable between liquid and CFU-gm cultures, particularly at earlier time points.
Conclusions:
- Liquid cultures provide a viable alternative for screening conditioned media for human CSF and assessing the cytotoxicity of agents to normal myeloid precursor cells.
- The behavior of CSF-responsive cells in liquid culture closely mirrors that of CFU-gm colony-forming cells.
- Liquid cultures enable both qualitative and quantitative evaluation of myeloid cell production and differentiation.