Ammonia effects in cultures of normal and transformed 3T3 cells

✍ Scribed by W. J. Visek; G. M. Kolodny; P. R. Gross

• Publisher: John Wiley and Sons
• Year: 1972
• Tongue: English
• Weight: 776 KB
• Volume: 80
• Category: Article
• ISSN: 0021-9541
• DOI: 10.1002/jcp.1040800308

✦ Synopsis

3T3 and SV-40 transformed 3T3 mouse fibroblasts were cultured i n media with serum and antibiotics plus ammonia (NH3 z NH,') added as NHaC1. Both cell lines cultured without added ammonia showed normal morphology and multiplication even though ammonia in the medium at the end of the culture period ranged from 35 to 48 ,ug/ml. Ammonia concentrations being significantly higher in media removed from cells at the end of the culture period than in media incubated identically without cells, verified that cells released substantial quantities of ammonia in addition to components of the medium which underwent spontaneous breakdown. Both cell lines showed changes i n morphology and highly significant reductions in cell multiplication which in- creased progressively as the concentration of added ammonia on the initial day of culture was increased to 35 ,ug/ml. Control 3T3 cultures released significantly greater quantities of ammonia per cell than control cultures of transformed cells but their multiplication was more adversely affected by added ammonia. There were downward shifts in pH o f t h e culturing medium for both cell lines as culture age increased at all concentrations of added ammonia. However, significant reductions in cell multiplication resulted from additions of ammonia that did not produce significant changes in extracellular pH. The data show that studies upon the effects of pH of the medium on cultured cells require control of ammonia concentrations.