Microfluidic cell culture and its application in high-throughput drug screening: cardiotoxicity assay for hERG channels.
Evaluation of drug cardiotoxicity is essential to the safe development of novel pharmaceuticals. Assessing a compound's risk for prolongation of the surface electrocardiographic QT interval and hence risk for life-threatening arrhythmias is mandated before approval of nearly all new pharmaceuticals. QT prolongation has most commonly been associated with loss of current through hERG (human ether-a-go-go related gene) potassium ion channels due to direct block of the ion channel by drugs or occasionally by inhibition of the plasma membrane expression of the channel protein. To develop an efficient, reliable, and cost-effective hERG screening assay for detecting drug-mediated disruption of hERG membrane trafficking, the authors demonstrate the use of microfluidic-based systems to improve throughput and lower cost of current methods. They validate their microfluidics array platform in polystyrene (PS), cyclo-olefin polymer (COP), and polydimethylsiloxane (PDMS) microchannels for drug-induced disruption of hERG trafficking by culturing stably transfected HEK cells that overexpressed hERG (WT-hERG) and studying their morphology, proliferation rates, hERG protein expression, and response to drug treatment. Results show that WT-hERG cells readily proliferate in PS, COP, and PDMS microfluidic channels. The authors demonstrated that conventional Western blot analysis was possible using cell lysate extracted from a single microchannel. The Western blot analysis also provided important evidence that WT-hERG cells cultured in microchannels maintained regular (well plate-based) expression of hERG. The authors further show that experimental procedures can be streamlined by using direct in-channel immunofluorescence staining in conjunction with detection using an infrared scanner. Finally, treatment of WT-hERG cells with 5 different drugs suggests that PS (and COP) microchannels were more suitable than PDMS microchannels for drug screening applications, particularly for tests involving hydrophobic drug molecules.
Department of Biomedical Engineering and Wisconsin Institutes for Medical Research, University of Wisconsin-Madison, Madison, WI, USA.
Xiaojing
X
Su
Edmond W K
EW
Young
Heather A S
HA
Underkofler
Timothy J
TJ
Kamp
Craig T
CT
January
David J
DJ
Beebe
D018929:Cell Culture Techniques; D004353:Drug Evaluation, Preclinical / Q000379:methods*; D051638:Ether-A-Go-Go Potassium Channels / Q000037:antagonists & inhibitors* / Q000378:metabolism; D057809:HEK293 Cells; D057166:High-Throughput Screening Assays; D006801:Humans; D017136:Ion Transport / Q000187:drug effects; D008133:Long QT Syndrome / Q000139:chemically induced; D044085:Microfluidics / Q000379:methods; D011108:Polymers / Q000737:chemistry; D026902:Potassium Channel Blockers / Q000009:adverse effects / Q000494:pharmacology
D051638:Ether-A-Go-Go Potassium Channels; D011108:Polymers; D026902:Potassium Channel Blockers