Merck, S5666-5ML, SPITE Medium Supplement (100×)

Application
For suspension cell cultures
Serum-free and reduced serum formulations often require purified forms of the factors normally supplied by serum that are suitable for in vitro growth and differentiation studies. These important factors include insulin, transferrin, selenium, pyruvate, and ethanolamine.

INSULIN is a polypeptide hormone that promotes the uptake of glucose and amino acids and may owe observed mitogenic effect to this property.

TRANSFERRIN is an iron-transport protein. Iron is an essential trace element, but can be toxic in the free form. To nourish cells in culture, it is supplied bound to transferrin in serum.

SELENIUM is an essential trace element normally provided by Serum.

SODIUM PYRUVATE has been shown to be beneficial as an additional energy source in some instances.

ETHANOLAMINE is a fatty acid that plays a significant role in the proliferation of hybridoma cells and frequently is added to supplements used for culturing these cells.

Nutritional studies indicate that the supplement components described are utilized by most mammalian cells. They enhance cell proliferation and decrease the serum requirement of many cell types. When the provided supplement is used with 2 to 4 percent serum, proliferation is reported to be similar to medium supplemented with 10 percent serum.
Preparation Note
Prepared in Earle′s Balanced Salt Solution (EBSS) without phenol red.
Other Notes
Contains 1.0mg/ml recombinant human insulin, 0.55mg/ml human transferrin (substantially iron-free), 0.5μg/ml sodium selenite, 12mg/ml pyruvate and 0.2mg/ml ethanolamine at the 100x concentration.
SPITE is a mixture of recombinant human insulin, human transferrin (partially iron-saturated), sodium selenite, sodium pyruvate and ethanolamine. It is designed for cell cultures in which media without sodium pyruvate are used.