Animal-Origin-Free ITS-G 100X Supplement

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• Bovine Serum in Cell CultureBovine serum is the liquid fraction of clotted cattle blood, added to culture medium at 5-10% to supply growth factors, hormones, transport and attachment proteins, lipids and trace elements that basal media do not contain. Fetal bovine serum (FBS) and fetal calf serum (FCS) are two names for the same product, collected from the fetus at slaughter of pregnant cows; newborn calf serum comes from calves under about 20 days old and donor bovine serum from controlled donor herds aged roughly 12-36 months, both containing more immunoglobulin and fewer growth factors than FBS. Because serum is an undefined biological material with substantial lot-to-lot variation, unresolved animal welfare questions and a volatile supply chain, defined serum-free and animal-origin-free media are increasingly preferred where the cell line will tolerate them.
• Recombinant Human Insulin in Cell Culture: Working Concentrations and ITSRecombinant human insulin is a 51-amino-acid, 5.8 kDa two-chain protein used as a core supplement in serum-free and chemically defined culture media, where it drives glucose and amino acid uptake, protein and lipid synthesis, and cell survival. It is produced in *E. coli* or yeast rather than extracted from animal pancreas, making it animal-origin-free, and is usually supplied as a zinc-stabilised dry powder. The typical working concentration is 5-10 ug/mL, the level delivered by a standard 1X ITS supplement, which is roughly a thousandfold above physiological insulin levels -- at that concentration insulin acts substantially through the IGF-1 receptor as well as its own. Because insulin is poorly soluble near neutral pH, powder must be dissolved in dilute hydrochloric or acetic acid before dilution into medium.