Chinese Hamster Ovary (CHO) cells are the main expression system for production of therapeutic recombinant (r-) proteins. The increased demand for these therapeutics has boosted the development of more robust production processes. Use of optimised feed/media has dramatically improved the performance of CHO cells in culture. However, such progress in biomass synthesis and r-protein production often come with an increased accumulation of lactate. In this study, we present a combined feeding strategy that uses galactose and lactate to replace glucose in CHO cell cultures. Replacement of glucose by galactose and lactate sustained cell growth and r-protein production in CHO cells. This strategy supported a better-balanced and more efficient metabolism, observed by an overall decreased consumption of carbon sources and amino acids, associated with an increased ATP production per C-mol consumed. Our results provide new insights of CHO cell metabolism in glucose-free media based on galactose and lactate. and glutamate, respectively, led to a significantly decreased rate of lactate production (C. Altamirano et al., 2000). However, with the implementation of this strategy, cell growth was dramatically decreased. In a subsequent study, a biphasic culture strategy using glucose and 4 galactose as primary and secondary carbon sources was designed to overcome the growth limitations observed in wholly galactose-based cultures. While the initial phase was characterised by a rapid cell growth with high rates of glucose consumption and lactate production, the second phase, when glucose was depleted, was characterised by the consumption of both galactose and lactate (produced in the initial phase) (Altamirano et al., 2006; Sun et al., 2013). To understand this phenomenon, Wilkens et al., (2011) studied lactate metabolism in CHO cell batch cultures growing in glucose and galactose using a metabolic flux analysis approach. These authors suggested that the lactate switch (i.e., from production to consumption) occurred when cytosolic pyruvate concentration was insufficient for cells to support their energy requirements, and that lactate uptake emerges as an alternative to incorporate carbon into the TCA cycle and improve energy metabolism (Wilkens et al., 2011). Similar observations were made in two different CHO cell lines (Gray et al., 2012; Sun et al., 2013) and in a mouse myeloma cell line (NS0) (Mulukutla et al., 2012) that underwent a lactate switch. Whilst galactose supplementation had little impact on r-protein production in CHO cell cultures, it did alter r-protein glycosylation (
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
customersupport@researchsolutions.com
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
This site is protected by reCAPTCHA and the Google Privacy Policy and Terms of Service apply.
Copyright © 2025 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.