Generation of bovine intraspecies hybridomas with initial suppressed growth.
Bovine B-cell hybrids were generated by fusing pokeweed mitogen (PWM) activated normal bovine peripheral blood lymphocytes (PBL) with an aminopterin-sensitive bovine B-cell line BL20. The fusion partner was derived by irradiation and growth in medium containing increasing concentrations of 6-thioguanine (6TG). Resultant cells were designated ATS/BL20. Polyethylene glycol-induced (PEG) fusion was used and hybrids were selected in hypoxanthine-aminopterin-thymidine (HAT) medium. Hybrid cell growth was noticed after 4 to 6 weeks of fusion following a period of quiescence. Hybrid formation was confirmed by selection in HAT medium, expression of cytoplasmic IgM (cIgM) and surface antigens and karyotype analysis.
Olobo
J O
JO
International Laboratory for Research on Animal Diseases, Nairobi, Kenya.
Black
S J
SJ
D000630:Aminopterin / Q000494:pharmacology; D000818:Animals; D000954:Antigens, Surface / Q000032:analysis; D001402:B-Lymphocytes / Q000166:cytology* / Q000276:immunology; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002470:Cell Survival / Q000187:drug effects; D003470:Culture Media / Q000494:pharmacology; D006822:Hybrid Cells / Q000166:cytology* / Q000276:immunology; D007074:Immunoglobulin G / Q000032:analysis; D007075:Immunoglobulin M / Q000032:analysis; D007621:Karyotyping; D009154:Mutation
D000954:Antigens, Surface; D003470:Culture Media; D007074:Immunoglobulin G; D007075:Immunoglobulin M; D000630:Aminopterin