Dielectrophoretic assembly of insulinoma cells and fluorescent nanosensors into three-dimensional pseudo-islet constructs.
Dielectrophoretic forces, generated by radio-frequency voltages applied to micromachined, transparent, indium tin oxide electrodes, have been used to condense suspensions of insulinoma cells (BETA-TC-6 and INS-1) into a 10 x 10 array of three-dimensional cell constructs. Some of these constructs, measuring approximately 150 microm in diameter, 120 microm in height and containing around 1000 cells, were of the same size and cell density as a typical islet of Langerhans. With the dielectrophoretic force maintained, these engineered cell constructs were able to withstand mechanical shock and fluid flow forces. Reproducibility of the process required knowledge of cellular dielectric properties, in terms of membrane capacitance and membrane conductance, which were obtained by electrorotation measurements. The ability to incorporate fluorescent nanosensors, as probes of cellular oxygen and pH levels, into these 'pseudo-islets' was also demonstrated. The footprint of the 10 x 10 array of cell constructs was compatible with that of a 1536 microtitre plate, and thus amenable to optical interrogation using automated plate reading equipment.
Pethig
R
R
BioCurrents Research Centre, Program in Molecular Physiology, Marine Biological Laboratory, Woods Hole, MA 02543, USA. ron@informatics.bangor.ac.uk
Menachery
A
A
Heart
E
E
Sanger
R H
RH
Smith
P J S
PJ
D018929:Cell Culture Techniques; D015194:Diffusion Chambers, Culture; D004553:Electric Conductivity; D046209:Electrophoresis, Microchip / Q000295:instrumentation / Q000379:methods*; D005456:Fluorescent Dyes; D006863:Hydrogen-Ion Concentration; D007340:Insulinoma / Q000378:metabolism / Q000473:pathology; D007515:Islets of Langerhans / Q000166:cytology / Q000378:metabolism; D008564:Membrane Potentials; D008839:Microelectrodes; D053769:Nanocapsules / Q000737:chemistry; D036103:Nanotechnology / Q000379:methods*; D010100:Oxygen / Q000032:analysis; D010190:Pancreatic Neoplasms / Q000378:metabolism / Q000473:pathology; D011166:Porphyrins / Q000737:chemistry; D033081:Shear Strength; D013314:Stress, Mechanical; D023822:Tissue Engineering / Q000379:methods; D054457:Tissue Scaffolds
D005456:Fluorescent Dyes; D053769:Nanocapsules; D011166:Porphyrins; D010100:Oxygen