Multiphenotypic whole-cell sensor for viability screening.
Here, we describe the fabrication of whole mammalian cell biosensors for the optical monitoring of cell viability. Three phenotypes were examined for their response to the addition of two model chemotoxins: sodium hypochlorite and sodium azide, and one model biotoxin, concanavalin A. Two sensing platforms containing cells, hydrogel microspheres, or hydrogel arrays, were also explored. Step changes in viability in response to small doses of sodium hypochlorite were seen nearly instantaneously in all cell lines, in solution, microspheres, and microarrays. Linear detection of sodium azide by entrapped hepatocytes was 0-10 microM, whereas the linear detection range for macrophages and endothelial cells was 0-75 microM. Macrophages and hepatocytes have a greater sensitivity, as indicated by a 40% change in fluorescence over the linear range, whereas endothelial cells show only a 15% change in fluorescence over the linear range. Using photoreaction injection molding, we were also able to generate a multiphenotype sensor that enables the measurement of the toxic effect of 100 microg/mL concanavalin A on macrophages and hepatocytes, but not on endothelial cells.
Itle
Laura J
LJ
Department of Chemical Engineering and The Huck Institute for the Life Sciences, The Pennsylvania State University, University Park, Pennsylvania 16802-4420, USA.
Pishko
Michael V
MV
D000818:Animals; D015374:Biosensing Techniques / Q000295:instrumentation* / Q000379:methods; D002470:Cell Survival; D002478:Cells, Cultured; D003208:Concanavalin A / Q000633:toxicity; D042783:Endothelial Cells / Q000187:drug effects; D005452:Fluoresceins; D022781:Hepatocytes / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects; D051379:Mice; D008863:Microspheres; D011092:Polyethylene Glycols; D019810:Sodium Azide / Q000633:toxicity; D012973:Sodium Hypochlorite / Q000633:toxicity
D005452:Fluoresceins; D003208:Concanavalin A; C085925:calcein AM; D011092:Polyethylene Glycols; D019810:Sodium Azide; D012973:Sodium Hypochlorite