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Journal of Nutrition Vol. 118 No. 12 December 1988, pp. 1487-1494
Copyright © 1988 by American Society for Nutrition
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Demonstration of a Choline Requirement for Optimal Keratinocyte Growth in a Defined Culture Medium1

Philip R. Gordon, Liebe K. Gelman and Barbara A. Gilchrest

USDA Human Nutrition Research Center on Aging, Tufts University, Boston, MA 02111

Nutrient requirements for proliferation and differentiated function of individual cell types can be determined using cell culture methodologies. Human epidermal keratinocytes are stimulated to grow by choline supplementation in the presence of myo-inositol when grown in a commercial nutrient medium containing six other defined supplements. The optimal range of choline concentrations varied among donor cell lines, but consistently fell between 36 µM and 180 µM. Addition of 72 µM choline increased cell yield to 250 ± 38% of that produced by myo-inositol supplementation alone and 92 ± 8% of that produced by addition of a highly mitogenic hypothalamic extract, which was previously required for good growth in this culture system. Supplementation of the basal medium with both the extract and choline resulted in 165 ± 13% of the cell yield observed with the extract addition alone. Supplementation with other phospholipid precursors did not further increase keratinocyte growth. Neither dermal fibroblasts nor epidermal melanocytes were stimulated by supplementation with choline, suggesting the high keratinocyte requirement is unusual. This completely defined culture medium for keratinocyte growth should prove useful in analyzing the role of phospholipids and other nutrients in human epidermis.

KEY WORDS: • keratinocyte • choline • growth • inositol

1 This work was supported by USDA Contract No. 53-3K06-5-10.

Manuscript received 16 December 1987. Revision accepted 25 August 1988.






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