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Characterization of serotonin transporter function in immortalized human lymphoblastoid cell lines
Author(s) -
HillKapturczak Nathalie,
Dougherty Donald M,
Galindo David,
Javors Martin
Publication year - 2011
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.25.1_supplement.657.6
Immortalized human lymphoblastoid cell lines (LCLs) are a model to study the expression, distribution, and function of serotonin transporters (SERT) without the confounds associated with platelets and lymphocytes. Differences in assay parameters exist in the few published studies of SERT function (5‐HT uptake) in LCLs. For example, cell counts range from 10 4 cells to 10 7 cells and assay buffers and incubation times vary significantly. We have optimized the 5‐HT uptake assay conditions for LCLs. Furthermore, we compared SERT function of LCLs grown in two different growth media: RPMI 1640 supplemented with 2 mM L ‐glutamine, 100 μg/100 U penicillin–streptomycin, and either 15% undialyzed serum or 15% dialyzed serum. Also, we observed that SERT function in LCLs is affected by 5‐HT in the growth medium. The concentrations of 5‐HT in the medium containing undialyzed serum versus dialyzed serum were 1.2 μM and 71.6 nM, respectively (HPLC‐EC). LCLs grown in undialyzed medium have lower V max values compared to LCLs grown in dialyzed medium (30.8 and 131 fmoles/10 7 cells‐min, respectively) without differences in K m values. Finally, kinetic parameters for 5‐HT uptake appear to be consistent over several changes of growth media. Our preliminary experiments provide important details for the consistent measurement of SERT function in a human cell model. Funding: NIH R01‐MH07768403S2.