Mathematical descriptions of hybridoma culture kinetics: II. The relationship between thiol chemistry and the degradation of serum activity.
The growth rate of the hybridoma cell line ATCC-CRL-1606 in low serum medium declines rapidly with time after inoculation. To characterize this phenomenon, the stability of the growth-promoting activity of serum was investigated. The activity of serum was found to de grade with time, and was stabilized by alterations in the medium formulation that acted to lower the oxidation/reduction potential. This included both the addition of thiols and the elimination of disulfides from the medium. Additionally, cysteine and other thiols were shown to stimulate growth in low serum, low cell density cultures, suggesting that thiols may be rate-limiting in low serum medium. Stimulation of growth by thiol addition was less significant at high cell levels, implying that the cells themselves may be acting to reduce their environment. A hypothesis is presented based on these results which suggests that the actual rate-limiting moieties in low serum cultures may be dithiols.
Glacken
M W
MW
Department of Applied Biological Sciences, Massachusetts Institute of Technology, Cambridge, MA 02139, USA.
Adema
E
E
Sinskey
A J
AJ