Abstract
Mammalian cells are generally considered to be unable to utilize polysaccharides for cell growth because the phospholipid bilayer in the cell membrane has very low permeability to sugars. With the recent discovery of the only known animal disaccharide transporter, a sucrose transporter, we considered the potential use of polysaccharides as energy source, because that can impact biopharmaceutical manufacturing by potentially increasing carbohydrate loading in the culture medium and decreasing lactate accumulation. In this study, we found that mammalian cells can utilize maltose for growth in the absence of glucose and successfully adapted CHO-K1, CHO-DG44 and HEK293 cells to grow in glucose-free, maltose-containing serum-free protein-free media. We then cultivated a non-adapted CHO-K1 producer cell line in media containing both glucose and maltose to show that the cells can utilize maltose in a biphasic manner, that maltose enters the cells, and that maltose utilization only took place in the presence of the cells. This is the first report of a protein-free mammalian cell culture using a disaccharide as energy source.
Highlights
The culture can be a model to elucidate yet unknown mechanisms of polysaccharide transport and metabolism in mammalian cells, such as the recent discovery of the first known animal sucrose transporter in Drosophila melanogaster[13]
To evaluate the use of disaccharides to support the growth of mammalian cells, a CHO cell line, CHO-K1, was cultivated using a seeding cell density of 0.3 × 106 cells/ml, with 3.6 g/l of maltose, sucrose, lactose, trehalose or glucose as energy source in a serum-free protein-free cell culture medium HyQ PF-CHO
We demonstrated that mammalian cell lines (CHO-K1, CHO-DG44 and Human Embryonic Kidney 293 (HEK293)) can be adapted to grow in maltose-supplemented serum-free protein-free media without glucose
Summary
The culture can be a model to elucidate yet unknown mechanisms of polysaccharide transport and metabolism in mammalian cells, such as the recent discovery of the first known animal sucrose transporter in Drosophila melanogaster[13]. We evaluated the use disaccharides, the simplest polysaccharides, to support the growth of a mammalian cell line in a serum-free protein-free culture. CHO and Human Embryonic Kidney 293 (HEK293) cells were chosen for the potential practical application of the study, since these are two of the most widely used mammalian cell line for the manufacture of recombinant protein therapeutics. The growth of these cells in disaccharide-containing media was characterized
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