A simple and improved method to generate human hybridomas.
By improving our previous method for production of human hybridomas, we developed a simple and remarkably efficient method for production of human hybridomas. We reformed our previous method in the following three points: (1) we added irradiated (30 Gy) myeloma cells as feeder cells to culture of fusion cells; (2) ouabain concentration in the selective medium was reduced from 2 x 10(-6) M to 1 x 10(-6) M; (3) selective medium (GIT-HAT-OL) was added to the culture after overnight cultivation of fusion cells. Consequently, we obtained higher fusion frequency (1/700 vs. 1/5500) compared with our previous method. By our present method, only so small number of human B cells (EBV-LCL) is required, so that the time necessary to establish human hybridomas is reduced.
Kudo
T
T
Cancer Cell Repository, Tohoku University, Sendai, Japan.
Saeki
H
H
Tachibana
T
T
D002459:Cell Fusion; D002478:Cells, Cultured; D002999:Clone Cells; D003470:Culture Media; D006801:Humans; D006822:Hybrid Cells; D006825:Hybridomas / Q000276:immunology*; D007158:Immunologic Techniques; D066298:In Vitro Techniques; D012217:Rheumatoid Factor / Q000276:immunology
D003470:Culture Media; D012217:Rheumatoid Factor