Journal of Cell Science, Vol 9, 529-537, Copyright © 1971 by Company of Biologists

Revised on March 3, 1971

The Glucose, Insulin and Glutamine Requirements of Suspension Cultures of HeLa Cells in a Difined Culture Medium

¹ Microbiology Department, Queen Elizabeth College (University of London), Campden Hill, London, W.8, England

The serum supplement in a defined medium for the growth of HeLa cells could be replaced by protamine-zinc-insulin (0.2 u./ml). Insulin (0.4 u./ml) replaced the growth-stimulatory properties of protamine-zinc-insulin, whilst protamine sulphate (5 µg/ml) was found to be toxic to the cells. The addition of insulin to cultures depleted of insulin increased both cell growth rates and maximum cell populations. In the defined medium, HeLa cells could only utilize glutamate when a small amount of glutamine was included. Glucose, at a level of 2 mg/ml, was shown to limit maximum cell populations. The growth yield from glucose was 295 µg cell dry weight/mg glucose. When the medium glucose concentration was increased to 4 mg/ml, HeLa cell populations in excess of 16 x 10⁵ cells (i.e. 640 µg dry weight)/ml were routinely achieved in the defined medium supplemented with insulin. Growth is then limited by the amino acid supply. Increasing the amino acid concentration of the medium by 50% raised the maximum cell population to 23.5x10⁵ cells (i.e. 940 µg dry weight)/ml.

This article has been cited by other articles:

				W. Arathoon and Birch JR Large-scale cell culture in biotechnology Science, June 13, 1986; 232(4756): 1390 - 1395. [Abstract] [PDF]