Selection of mouse X hamster hybrids using HAT medium and a polyene antibiotic.
In the present study we tested the feasibility of utilizing a structurally modified polyene antibiotic, amphotericin B methyl ester (AME), as a half-selection agent for isolating somatic cell hybrids. By using HAT medium supplemented with AME we have isolated interspecific mouse-hamster hybrids from mixed cultures of mouse (TK-C1 ID or HPRT-A9) and hamster (BHK/C 13) cells fused with Sendai virus, lysolecithin or polyethylene glycol. Hybrid cells proliferated and clones were isolated after 2 to 3 weeks growth in three changes of HAT-AME medium and subsequent growth in HAT medium alone. In contrast, genetically deficient parental C1 1D or A9 cells and AME-sensitive BHK/C 13 cells were killed using a similar growth protocol. The described technique is simple, efficient and permits one to use a cell line without a genetic defect in combination with a genetically deficient cell type in hybrid formation.
D000630:Aminopterin / Q000494:pharmacology; D000666:Amphotericin B / Q000031:analogs & derivatives* / Q000494:pharmacology; D000818:Animals; D002478:Cells, Cultured / Q000187:drug effects; D006224:Cricetinae; D003470:Culture Media; D004121:Dimethyl Sulfoxide / Q000494:pharmacology; D005821:Genetic Techniques; D006822:Hybrid Cells / Q000187:drug effects*; D007042:Hypoxanthines / Q000494:pharmacology; D007621:Karyotyping; D051379:Mice; D008858:Microscopy, Phase-Contrast; D013936:Thymidine / Q000494:pharmacology
D003470:Culture Media; D007042:Hypoxanthines; C017501:methylamphotericin B; D000666:Amphotericin B; D000630:Aminopterin; D013936:Thymidine; D004121:Dimethyl Sulfoxide