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An improved method for selecting T-cell hybridomas by the fluorescence-activated cell sorter
Journal of Immunological Methods
|January 1, 1979
Summary
This study presents a two-step fluorescence-activated cell sorting (FACS) method to isolate T cell hybridomas. This technique efficiently selects hybrid cell lines from fused T cells and lymphoma cells.
Area of Science:
- Immunology
- Cell Biology
- Biotechnology
Background:
- T cell hybridomas are crucial tools in immunology research.
- Efficient selection methods are needed to isolate these hybrid cells from fusion products.
Purpose of the Study:
- To develop and describe a novel two-step sorting procedure for selecting T cell hybridomas.
- To improve the efficiency of hybridoma isolation using fluorescence-activated cell sorting (FACS).
Main Methods:
- A two-step FACS procedure was employed.
- Initial sorting based on cell-surface antigen differences.
- Secondary sorting utilized DNA content analysis with Hoechst 33258 dye after in-vitro culture.
Main Results:
- The described FACS method successfully isolated T cell hybridomas.
- Chromosome analysis confirmed 15-25% of resulting clones were hybrid lines.
- The procedure effectively enriches for hybrid cell populations.
Conclusions:
- The two-step FACS sorting procedure is an effective method for selecting T cell hybridomas.
- This technique offers a reliable approach for generating hybridoma cell lines for immunological studies.
- The combination of antigen and DNA content sorting enhances hybridoma isolation efficiency.