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Selection of mouse X hamster hybrids using HAT medium and a polyene antibiotic
In Vitro
|February 1, 1978
Summary
Amphotericin B methyl ester (AME) effectively isolates somatic cell hybrids. This method allows using a normal cell line with a deficient one for hybrid formation, proving simple and efficient.
Area of Science:
- Cell Biology
- Genetics
- Biochemistry
Background:
- Somatic cell hybridization is crucial for genetic studies.
- Existing selection methods for hybrid cells can be limiting.
Purpose of the Study:
- To evaluate amphotericin B methyl ester (AME) as a half-selection agent for isolating somatic cell hybrids.
- To develop a simplified and efficient method for generating interspecific hybrids.
Main Methods:
- Utilized HAT medium supplemented with AME for selection.
- Fused mouse (TK-C1 ID or HPRT-A9) and hamster (BHK/C 13) cells using Sendai virus, lysolecithin, or polyethylene glycol.
- Cultured hybrid cells in HAT-AME medium followed by HAT medium alone.
Main Results:
- Successfully isolated interspecific mouse-hamster hybrid cells.
- Demonstrated selective killing of parental cells (genetically deficient mouse cells and AME-sensitive hamster cells) while hybrid cells proliferated.
- Achieved hybrid cell proliferation and clone isolation within 2-3 weeks.
Conclusions:
- AME is a feasible and effective half-selection agent for somatic cell hybrid isolation.
- This technique simplifies hybrid formation by allowing the use of a non-genetically deficient cell line.
- The method is efficient and robust for generating interspecific hybrids.