CNRS, Laboratoire Réactions et Génie des Procédés, UPR- 3349, 2 avenue de la forêt de Haye, 54505, Vandœuvre-lès-Nancy, France.
Cytotechnology. 2013 Aug;65(4):629-41. doi: 10.1007/s10616-012-9519-1. Epub 2012 Dec 14.
Many studies underlined the great benefits of hydrolysates used as additives in animal free media on cell culture performances. However, to precisely define hydrolysate supplementation strategies, a deeper understanding of their effect on cell growth and protein production is required. In the present study, the effect of addition of one yeast extract (YE) and two yeast peptones (named YP.A and YP.B) in a chemically defined medium was first assessed on cell culture performances. Interestingly, specific effects were found depending on the degree of degradation of yeast hydrolysates. The YE at 1 g L(-1) increased the maximal cell density by 70 %, while a mixture of YE (1 g L(-1)) and YP.A (4 g L(-1)) increased IgG production by 180 %. These conditions were then evaluated on the CHO cell kinetics all over cultures. Hydrolysates extended the cell growth phase in Erlenmeyer flask and increased the maximal growth rate in bioreactor up to 20 %. Cell growth stimulation induced by hydrolysates addition was linked with energetic metabolism improvement suggesting that they promote oxidative pathway. Furthermore, hydrolysates provided an additional source of substrate that supported cell growth despite glutamine limitation.
许多研究强调了水解产物作为添加剂在无动物培养基中对细胞培养性能的巨大益处。然而,为了准确定义水解产物的补充策略,需要更深入地了解它们对细胞生长和蛋白质生产的影响。在本研究中,首先评估了在化学定义培养基中添加一种酵母提取物(YE)和两种酵母肽(分别命名为 YP.A 和 YP.B)对细胞培养性能的影响。有趣的是,发现具体的影响取决于酵母水解产物的降解程度。在 1 g L(-1)的 YE 提高了最大细胞密度 70%,而 YE(1 g L(-1))和 YP.A(4 g L(-1))的混合物提高了 IgG 产量 180%。然后在整个培养过程中对 CHO 细胞动力学进行了这些条件的评估。水解产物延长了摇瓶中的细胞生长阶段,并将生物反应器中的最大生长速率提高了 20%。水解产物添加引起的细胞生长刺激与能量代谢的改善有关,表明它们促进了氧化途径。此外,水解产物提供了额外的底物来源,即使在谷氨酰胺有限的情况下也能支持细胞生长。