Optimizing CHO hERG for automated patch clamping

著者: Søren Friis, Hervør L Olsen, Jeffrey Webber, M Knirke Jensen, Dorthe Nielsen, Morten Sunesen, Chris Mathes

Cultured cells from mammalian cell lines incubated at 37 °C can be used for automated patch clamp (APC), including compound screening, for only a relatively brief period of time. Usually this time period extends from the time of ~50 % confluence to full confluence. For CHO cells, the preferred cell type for APC, expressing hERG potassium channels the duration of the usable period is 1-2 days. We here report that incubating CHO-hERG cells at a reduced temperature, 30 °C, for 1-5 days prior to APC experiments is advantageous in several ways: (1) it increases the percentage of cells with acceptable hERG currents (> 50 pA), (2) it increases the average whole-cell current significantly, and (3) it
slows cell proliferation and extends the usable period up to at least 5 days. In addition, we have tested whether the altered conditions affect the IC50 obtained for known hERG inhibitors. For all experiments the automated patch clamp system QPatch 16 was used.