bioMérieux, Chemin de l'orme, Marcy L'etoile, F-69280, France.
Cytotechnology. 2003 May;42(1):1-11. doi: 10.1023/A:1026185615650.
The aim of this study was to understand the metabolism kinetics of Vero cells grown on microcarriers in bioreactors in serum-free medium (SFM). We sought to determine what nutrients are essential for Vero cells and how they are consumed. Contrary to glucose and to most of the amino acids, glutamine and serine were very quickly depleted in this medium and can be supposed to be responsible for cell apoptosis. Lactate and ammonium ions did not reach toxic levels for Vero cells. We payed more attention to the lactate metabolism. Usually we observed that after about 2 days lactate was consumed in serum-containing media, but its concentration plateaud in SFM. Moreover, the addition of serum in SFM provoked lactate consumption and the rate of glucose and glutamine consumption was twice as high as in the SFM not supplemented with serum. The depletion of glutamine and serine and the metabolic deviations leading to a shortage of intermediate products required for other metabolic pathways probably contribute to the lower cell yield and higher cell death rate in SFM.
本研究旨在了解无血清培养基(SFM)中微载体上生长的 Vero 细胞在生物反应器中的代谢动力学。我们试图确定 Vero 细胞所需的营养物质以及它们的消耗方式。与葡萄糖和大多数氨基酸不同,谷氨酰胺和丝氨酸在这种培养基中很快被耗尽,可能是导致细胞凋亡的原因。乳酸和铵离子没有达到 Vero 细胞的毒性水平。我们更关注乳酸的代谢。通常我们观察到,在含有血清的培养基中,大约 2 天后乳酸被消耗,但在 SFM 中其浓度达到平台期。此外,在 SFM 中添加血清会刺激乳酸的消耗,并且葡萄糖和谷氨酰胺的消耗速率是不添加血清的 SFM 的两倍。谷氨酰胺和丝氨酸的消耗以及导致其他代谢途径所需中间产物短缺的代谢偏差,可能导致 SFM 中的细胞产量较低和细胞死亡率较高。