Cell-to-cell variation in store-operated calcium entry in HEK-293 cells and its impact on the interpretation of data from stable clones expressing exogenous calcium channels.
To investigate the effect of cell-to-cell variation in store-operated calcium entry (SOCE) on the evaluation of data from stable cell clones selected following gene transfection, we measured SOCE in 2700 individual HEK-293 cells from the parent population and in 1900 individual cells from a clonal subpopulation of HEK-293 cells. We applied statistical resampling techniques to model conditions where one would compare the average SOCE in n control clones to the average SOCE in n experimental clones (n = 1-200). For an overexpression experiment with n = 1, there is a 27% chance of observing a 100% or higher difference in SOCE between clones, with n = 10 there is a 34% probability of observing a 20% or greater difference in SOCE, and with n = 100, there is less than a 10% chance of seeing a 10% or greater difference in SOCE, based solely on random selection of clones from the parent HEK-293 cell population. To assure that the degree of cell-to-cell variation was predictive of the degree of clone-to-clone variation, we measured SOCE in 270 clones, each arising from a single cell, and found the variation to be very similar to that observed for individual cells.
Babnigg
G
G
Department of Pharmacological and Physiological Sciences, University of Chicago, IL 60637, USA.
Heller
B
B
Villereal
M L
ML
D002118:Calcium / Q000378:metabolism*; D015220:Calcium Channels / Q000378:metabolism*; D000252:Calcium-Transporting ATPases / Q000037:antagonists & inhibitors; D002460:Cell Line; D002999:Clone Cells; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005456:Fluorescent Dyes / Q000378:metabolism; D006801:Humans; D007668:Kidney / Q000166:cytology* / Q000378:metabolism; D018002:Receptors, Bradykinin / Q000235:genetics / Q000378:metabolism; D019284:Thapsigargin / Q000494:pharmacology; D014162:Transfection
D015220:Calcium Channels; D004791:Enzyme Inhibitors; D005456:Fluorescent Dyes; D018002:Receptors, Bradykinin; D019284:Thapsigargin; D000252:Calcium-Transporting ATPases; D002118:Calcium