Department of Biological Sciences, Korea Advanced Institute of Science & Technology, 373-1 Guseong-Dong, Yuseong-gu, Daejeon, Republic of Korea.
J Biotechnol. 2011 Sep 10;155(2):225-31. doi: 10.1016/j.jbiotec.2011.06.020. Epub 2011 Jun 23.
The effect of growth factor (GF) and sodium butyrate (NaBu) on Chinese hamster ovary (CHO) cell growth, cell viability and antibody production was investigated using shaking flasks in GF-containing and GF-deficient medium containing 0, 1 and 3mM NaBu. The withdrawal of GF and the addition of NaBu suppressed cell growth, but they significantly increased specific antibody productivity, q(Ab). Interestingly, the withdrawal of GF in combination with the addition of NaBu markedly retarded cell death, leading to extended culture longevity. For instance, at 3mM NaBu, cell viability fell below 80% after day 4 in GF-containing medium, but it remained over 80% until day 18 in GF-deficient medium. Due to the enhanced q(Ab) and the extended culture longevity, approximately 2-fold increase in total antibody production was achieved in pseudo-perfusion culture with 1mM NaBu in GF-deficient medium, compared to the culture in GF-containing medium. The effect of GF and NaBu on the change in the expression and activity of cellular proteins, c-Myc, Bcl-2 and pyruvate dehydrogenase (PDH), was also investigated. Both the withdrawal of GF and the addition of NaBu decreased the expression of c-Myc. The expression of Bcl-2 was enhanced by the addition of NaBu in a dose-dependent manner while it was not affected by the withdrawal of GF. In addition, both the withdrawal of GF and the addition of NaBu reduced metabolic rates, q(Glc), q(Lac) and Y(Lac/Glc), and increased PDH activity while not affecting PDH expression, suggesting that they may reduce the glycolytic rates, but enhance the conversion rates of pyruvate to TCA intermediates. Taken together, the withdrawal of GF in combination with the addition of NaBu can be considered as a relevant strategy for alleviating NaBu-induced cell apoptosis and enhancing antibody production since it can be easily implemented as well as enhance q(Ab) and extend culture longevity.
研究了生长因子（GF）和丁酸钠（NaBu）对中国仓鼠卵巢（CHO）细胞生长、细胞活力和抗体产生的影响，方法是在含有 GF 和 GF 缺乏的培养基中使用摇瓶进行，培养基中含有 0、1 和 3mM 的 NaBu。GF 的去除和 NaBu 的添加抑制了细胞生长，但它们显著提高了特异性抗体的生产力，q(Ab)。有趣的是，GF 的去除与 NaBu 的添加相结合显著延缓了细胞死亡，从而延长了培养寿命。例如，在 3mM 的 NaBu 下，含 GF 的培养基中细胞活力在第 4 天低于 80%，但在 GF 缺乏的培养基中直到第 18 天仍保持在 80%以上。由于 q(Ab)的提高和培养寿命的延长，在 GF 缺乏的培养基中用 1mM NaBu 进行假性灌注培养时，与在含 GF 的培养基中相比，总抗体产量增加了约 2 倍。还研究了 GF 和 NaBu 对细胞蛋白 c-Myc、Bcl-2 和丙酮酸脱氢酶（PDH）表达和活性变化的影响。GF 的去除和 NaBu 的添加都降低了 c-Myc 的表达。NaBu 的添加以剂量依赖的方式增强了 Bcl-2 的表达，而 GF 的去除对其没有影响。此外，GF 的去除和 NaBu 的添加都降低了代谢率，q(Glc)、q(Lac)和 Y(Lac/Glc)，并增加了 PDH 活性，而不影响 PDH 表达，这表明它们可能降低糖酵解速率，但提高丙酮酸转化为 TCA 中间产物的速率。总之，GF 的去除与 NaBu 的添加相结合可以被认为是一种缓解 NaBu 诱导的细胞凋亡和提高抗体产生的相关策略，因为它既易于实施，又能提高 q(Ab)并延长培养寿命。
