Observations on the use of a cation exchange resin for the preparation of metal-depleted media for lymphocyte culture.
A chelating resin specific for divalent cations (Chelex) was used to prepare metal-depleted media for lymphocyte culture. A batch procedure (resin in pH 7.4 phosphate buffer/specimen, 1:1) removed 70-80% of iron, 77-87% of copper and 88-98% of zinc, calcium and magnesium. At variance with other reports, when a resin/specimen ratio of 1:4 was used, iron chelation decreased to 40%, whereas other cation chelation remained unchanged. Best chelation for iron and calcium was obtained at pH 5-6.4; for copper, zinc and magnesium, at pH 7.4-8.0. During the procedure protein content decreased by 8-10%; arginine and lysine by 80%; asparagine, cystine, tyrosine and phenylalanine by 60%, other amino acids by 35%. These new data suggest that cation-depleted media prepared with Chelex may be used to study the effects of cations on lymphocytes in culture, provided that the most appropriate pH and resin/specimen ratio are selected and adequate amino acid replacement is performed. Results on normal human lymphocytes are reported.
Carpentieri
U
U
Department of Pediatrics, University of Texas Medical Branch, Galveston 77550.
Myers
J
J
Daeschner
C W
CW
Haggard
M E
ME
D002411:Cation Exchange Resins; D003470:Culture Media; D046508:Culture Techniques / Q000379:methods; D006801:Humans; D008214:Lymphocytes / Q000166:cytology*; D008670:Metals / Q000032:analysis*; D011137:Polystyrenes; D011145:Polyvinyls
D002411:Cation Exchange Resins; D003470:Culture Media; D008670:Metals; D011137:Polystyrenes; D011145:Polyvinyls; C006960:chelex