Online monitoring of BALB/3T3 metabolism and adhesion with multiparametric chip-based system.
A multiparametric chip-based system was employed to measure cell adhesion, metabolism, and response to metal compounds previously classified as cytotoxic in immortalized mouse fibroblasts (BALB/3T3 cell line). The system measures in parallel, online, and in label-free conditions the extracellular acidification rates (with pH-sensitive field effect transistors [ISFETs]), the cellular oxygen consumption (with amperometric electrode structures [Clark-type sensors]), and cell adhesion (with impedimetric interdigitated electrode structures [IDESs]). The experimental protocol was optimized to monitor metabolism and adhesion of the BALB/3T3 cell line. A total of 70,000 cells and a bicarbonate buffer-free running low-glucose Dulbecco's modified Eagle's medium (DMEM) containing 10% fetal clone serum III and 1mM Hepes were selected to maintain cells in good conditions on the chip during the measurements performed under perfusion conditions. Cells were exposed to sodium arsenite, cadmium chloride, and cis-platinum at concentrations ranging from 1 to 100 microM. The kinetics of cell response to these compounds was analyzed and suggests that the Clark-type sensors can be more sensitive than IDESs and ISFETs in detecting the presence of high chemical concentration when short exposure times (i.e., 2h) are considered. The cytotoxicity data obtained from the online measurements of acidification, respiration, and adhesion at 24h compare well, in terms of half-inhibition concentration values (IC(50)), with the ones obtained using 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) test and colony-forming efficiency (CFE) assay. The results show a good sensitivity of the system combined with the advantages of the online and label-free detection methods that allow following cell status before, during, and after the treatment in the same experiment.
Ceriotti
L
L
Nanotechnology and Molecular Imaging Unit, Institute for Health and Consumer Protection, Joint Research Centre, I-21020 Ispra, Italy.
Kob
A
A
Drechsler
S
S
Ponti
J
J
Thedinga
E
E
Colpo
P
P
Ehret
R
R
Rossi
F
F
D016475:3T3 Cells; D000818:Animals; D018053:Arsenites / Q000633:toxicity; D015374:Biosensing Techniques; D002021:Buffers; D002122:Calcium Chloride / Q000633:toxicity; D002448:Cell Adhesion / Q000187:drug effects; D002461:Cell Line, Transformed; D019069:Cell Respiration / Q000187:drug effects; D002945:Cisplatin / Q000633:toxicity; D003470:Culture Media / Q000737:chemistry; D004305:Dose-Response Relationship, Drug; D017097:Electric Impedance; D004563:Electrochemistry / Q000295:instrumentation; D004867:Equipment Design; D005347:Fibroblasts / Q000378:metabolism*; D006531:HEPES / Q000737:chemistry; D020128:Inhibitory Concentration 50; D007700:Kinetics; D056656:Lab-On-A-Chip Devices; D051379:Mice; D008807:Mice, Inbred BALB C; D008839:Microelectrodes; D009862:Online Systems; D010101:Oxygen Consumption / Q000187:drug effects; D015203:Reproducibility of Results; D012680:Sensitivity and Specificity; D012825:Silicon / Q000737:chemistry; D017670:Sodium Compounds / Q000633:toxicity; D013997:Time Factors; D018675:Toxicity Tests
D018053:Arsenites; D002021:Buffers; D003470:Culture Media; D017670:Sodium Compounds; C017947:sodium arsenite; D002122:Calcium Chloride; D002945:Cisplatin; D006531:HEPES; D012825:Silicon