HiDef-ITS Cell Culture Supplement

Defined Bioscience’s HiDef-ITS is a defined formulation designed for the routine maintenance and low-density attachment of adherent cells under reduced serum conditions. HiDef-ITS is a combination of insulin (to stimulate glucose uptake), transferrin (to facilitate iron transport), and sodium selenite (a potent antioxidant and common component supplied by serum), previously shown to reduce the amount of FBS needed in the culture of a wide range of cell types, including primary cells, immortalized cell lines, and stem cells. HiDef-ITS should be used with other growth factors, sera, or additives (NOT INCLUDED) depending on your cell culture application.

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Defined Bioscience’s HiDef-ITS is a defined formulation designed for the routine maintenance and low-density attachment of adherent cells under reduced serum conditions. HiDef-ITS is a combination of insulin (to stimulate glucose uptake), transferrin (to facilitate iron transport), and sodium selenite (a potent antioxidant and common component supplied by serum), previously shown to reduce the amount of FBS needed in the culture of a wide range of cell types, including primary cells, immortalized cell lines, and stem cells.

HiDef-ITS is provided as a 100x supplement in Earle’s Balanced Salt Solution (EBSS) without phenol red, and at this concentration consists of 1.00 mg/mL insulin, 0.55 mg/mL transferrin, and 0.67 µg/mL sodium selenite, all using animal origin-free componentry. Depending on your application, FBS can be reduced to 4% or less when maintaining cells in a medium containing HiDef-ITS.

• Insulin: Insulin promotes the uptake of glucose, amino acids, cations, and phosphates, stimulates cell growth and cell cycle progression, and offers anti-apoptotic and lipogenic effects.
• Transferrin: Transferrin serves as a high-affinity iron carrier and potent antioxidant, reducing toxicity associated with free iron exposure, and offers further protective benefits to cells in reduced-serum conditions.
• Sodium selenite: Sodium selenite additionally serves as an antioxidant, as well as a co-factor for seleno-enzymes.

HiDef-ITS 100x is provided in liquid form in 10-mL volumes, sufficient for 1 L of medium. This supplement can be used with a wide range of basal medium formulations, including Defined Bioscience’s fully defined DMEM/F12 basal medium (Catalog # LSB-101). Following initial adaptation and depending on user application, cells cultured in HiDef-ITS can thrive under reduced serum conditions.

Each lot of HiDef-ITS is used in combination with basal medium in performance testing in a culture assay evaluating fibroblast outgrowth and viability.