Jl. Goergen et al., DETERMINATION OF CELL-LYSIS AND DEATH KINETICS IN CONTINUOUS HYBRIDOMA CULTURES FROM THE MEASUREMENT OF LACTATE-DEHYDROGENASE RELEASE, Cytotechnology, 11(3), 1993, pp. 189-195
The death of the hybridoma VO 208 in a continuous culture at pH 7 and
6.8 was investigated by measuring both the appearance of visible dead
cells which do not exclude the trypan blue dye and the release of lact
ate dehydrogenase (LDH) in the culture medium. The intracellular LDH w
as found to be completely released either when live cells lysed or whe
n they were transformed into visible dead cells. No significant lysis
of blue dead cells could be observed at the two different pH. Using a
LDH balance over the culture system, cell lysis was found negligible a
t pH 7, but accounted for 20% of the total cell death at pH 6.8. A met
hodology is proposed to evaluate the rate constants of hybridoma lysis
and total death. For the investigated cell line in continuous culture
, the calculated total cell death rate constant was found to increase
from 0.002 h-1 to 0.01 h-1 when decreasing the pH from 7 to 6.8.