Requirements of Cultured Mammalian Cells for Vitamin B12 and Biotin
FORT DETRICK FREDERICK MD
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Eagle reported in 1955 and 1957 that eight vitamins were essential for the growth of most mammalian cells in culture. No evidence for requirement of vitamin B12 or of biotin was presented. Sanford and co-workers in 1963 and 1964 reported evidence for requirement of both of these vitamins in certain cell strains, but no quantitative data were presented. In 1967, Ham reported that 3 x 10 to the minus 10th power M biotin permitted successful cloning of a Chinese hamster cell line. We have found that both HeLa and L cells required approximately 10 to the minus 11th power to 10 to the minus 10th power M vitamin B12 for optimal growth in a chemically defined medium. Requirement of HeLa cells for biotin was demonstrated initially with avidin, a biotin inactivator. The inhibitory activity of avidin on growth of HeLa cells was reversible by addition of biotin. Serial passage of both HeLa and L cells in presumably biotin-free medium resulted in achieving biotin-deficient cells. Both cell lines required approximately 10 to the minus 8th power M biotin for optimal growth in a chemically defined growth medium.
- Anatomy and Physiology