Abstract
Abstract: :
Purpose: To study the effect of L–glutamine on the growth and melanogenesis of cultured uveal melanocytes (UM).Methods: Human UM were isolated and cultured from donor eyes as previously reported. Early passages of UM were plated into multi–well plates and cultured with glutamine–free FIC medium (glutamine–free F12 medium with 20 ng/ml bFGF, 1 mM IBMX, 10 ng/ml cholera toxin and 10% serum). L–glutamine was added to the culture medium at various concentrations. Each test condition was performed in triplicate. Cells cultured with FIC medium without glutamine were used as controls. After 6 days, the cells were detached and counted. Their melanin content was measured spectrophotometrically. Results: UM cultured with glutamine–free medium grew slowly and produced less melanin in vitro. Glutamine at 1–5 mM significantly stimulated growth and melanogenesis in a dose–dependent manner (p < 0.01). The cell number and melanin content per well of the 5 mM glutamine group were 132% and 193% of the controls, respectively. Glutamine did not stimulate the growth and melanogenesis of UM at 10 mM. Conclusion: The optimal concentration of L–glutamine in the culture medium for the growth and melanogenesis is 3–5 mM. The glutamine concentration in the F12 medium is only 1 mM, which is inadequate for UM. Addition of 2–4 mM of glutamine to F12 medium is important for the optimal growth and melanogenesis of UM in vitro. CR: None. Supported by the New York Eye and Ear Infirmary Departments of Ophthalmology and Pathology Research Funds, New York, NY.
Keywords: melanocytes • uvea • melanoma