Chemically defined serum-free media for the cultivation of primary cells and their susceptibility to viruses

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Abstract
Chemically defined media SFRE-199-1 for the growth and SFRE-199-2 for the maintenance of primary baboon kidney (Bak) cell cultures were formulated by supplementing medium M199 with insulin, sodium pyruvate, zinc sulfate, and increasing arginine-HCl, cysteine, cystine,l-glutamine,l-glutamic acid, glycine, histidine, tyrosine, and glucose to maximally active nontoxic concentrations. For prolonged maintenance of the cells, physiological pH control, and blocking of excessive lactic acid accumulation in the spent medium of the cell cultures, it was necessary to supplement the medium containing Earle's balanced salts withd-(+) galactose. The cells grew and were maintained equally well on glass or polystyrene surfaces. Selenium, when added to growth medium or substituted for insulin and zinc sulfate, did not stimulate cell growth. Electron microscopy showed that numerous dense particles, approximately 250 to 400 Å in diameter, with the appearance of glycogen, were found throughout the cytoplasm in the cells grown in SFRE-199-1 and maintained in SFRE-199-2. Echovirus types 1 to 3, poliovirus types 1 to 3, coxsackievirus types B2, B4, B5,Herpesvirus hominis type 1, simian herpesvirusH. simiae and SA8, and simian adenovirus SV34 when titrated in primary Bak cells and grown and maintained in SFRE-199-1 and 2, respectively, developed titers comparable to those obtained in conventionally grown and maintained cells.