Hamster ascites tumor cell line bhk 21/c.13/t.6/ascites useful in submerged serum-free agar cultures to support virus growth

摘要

Polyoma-modified BHK 21 cells are further modified to produce new cell strains or cell lines, designated ATCC No. CL 10.1 (BHK 21/C.13/T.6/ Ascites), having the property that they can be cultivated in agar, by inoculating the polyoma-modified BHK 21 cells into the hamster cheek pouch to form tumours, removing a tumour so formed and inoculating it into a hamster subcutaneously to form subcutaneous tumours, inoculating material from the subcutaneous tumours intraperitoneally into a hamster to form ascites tumours and recovering ascites tumour cells from the animal. Subcutaneously-formed tumours may be passaged by subcutaneous implantation in a hamster before the intraperitoneal inoculation. The procedure is described in detail. The cells are cultured in a gel overlay consisting of agar (less than 0.5%), Eagle/Hanks medium fortified to twice its normal strength of salts, vitamins and amino acids, tryptose phosphate, calf serum and aqueous sodium carbonate solution; the overlay is poured on to an under layer of similar composition except that the concentration of agar amounts to 0.9%. The ascites tumour cells grow well under submerged culture in a serum-free medium consisting of Eagle/ Hanks medium containing twice its normal strength of vitamins and amino acids, tryptose phosphate, yeast extract and aqueous sodium carbonate solution The ascites tumour cells may be employed for the cultivation of viruses (specified), especially those difficult to cultivate in the presence of serum. Mutants of the ascites tumour cells may be obtained by the agar-plate technique by the use of selective media.ALSO:Polyomo-modified BHK 21 cells are further modified to produce p new cell strains or cell lines, designated ATCC No. CL. 10.1 (BHK 21/C. 13/T. 6/Ascites), having the property that they can be cultivated in agar (see Division C6). The ascites tumour cells may be employed for the cultivation of the following viruses, especially those difficult to cultivate in the presence of serum: E.M.C, Semliki Forest, equine encephalomyelitis, Echo 10, Reoviruses II and III, Sendai, fowl plague, adenovirus type 5, rabies, herpes, influenza and vaccinia.