Goergen J L, Marc A, Engasser J M
Laboratoire des Sciences du Génie Chimique, Institut National Polytechnique de Lorraine, CNRS, Nancy, France.
Cytotechnology. 1993;11(3):189-95. doi: 10.1007/BF00749869.
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 lactate dehydrogenase (LDH) in the culture medium. The intracellular LDH was found to be completely released either when live cells lysed or when 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 at pH 7, but accounted for 20% of the total cell death at pH 6.8. A methodology 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.
通过测量不排斥台盼蓝染料的可见死细胞的出现以及培养基中乳酸脱氢酶（LDH）的释放，研究了杂交瘤VO 208在pH值为7和6.8的连续培养中的死亡情况。发现当活细胞裂解或转化为可见死细胞时，细胞内的LDH会完全释放。在两种不同的pH值下，未观察到蓝色死细胞有明显的裂解现象。通过对培养系统进行LDH平衡分析，发现在pH值为7时细胞裂解可忽略不计，但在pH值为6.8时，细胞裂解占总细胞死亡的20%。提出了一种评估杂交瘤裂解和总死亡速率常数的方法。对于连续培养中所研究的细胞系，当pH值从7降至6.8时，计算得出的总细胞死亡率常数从0.002 h⁻¹增加到0.01 h⁻¹。
