Τίτλος:
Development of a novel, multi-analyte biosensor system for assaying cell
division: Identification of cell proliferation/death precursor events
Γλώσσες Τεκμηρίου:
Αγγλικά
Περίληψη:
A novel, miniaturized biosensor system was created by combining the
electrophysiological response of immobilized cells with
superoxide-sensing technology, optical and fluorescence microscopy. Vero
cells were immobilized in a calcium alginate matrix (at a density of 1.7
x 10(6) cells ml(-1)). A 0.5 cm x 0.5 cm piece of cell-containing gel
matrix was aseptically adhered on a glass microscope slide with a
microfabricated gold electrode array, sealed with a cover slip and
provided with Dulbecco’s medium +10% (v/v) fetal calf serum every day
by means of a capillary feeding tube. During a culture period of 7 days,
the membrane potential of immobilized cells was continuously monitored,
while cell division was assayed with an optical microscope. In addition,
daily measurements of immobilized cell membrane potential, viability,
RNA and calcium concentration, radical oxygen species (ROS) and
glutathione accumulation, were conducted by fluorescence microscopy
after provision of an appropriate dye. Superoxide accumulation was
assayed by covering the electrodes with superoxide dismutase (SOD).
Maximum cell membrane potential values and superoxide production were
observed upon initiation of cell division. Using the novel biosensor, we
were able to correlate seven different cell physiological parameters to
each other and formulate a model for ROS-mediated signaling function on
cell division and death. In addition, we were able to predict cell
proliferation or death by comparing the relative response of the
electrophysiological and superoxide sensor during the culture period.
(c) 2005 Elsevier B.V. All rights reserved.
Συγγραφείς:
Kintzios, S
Marinopoulou, I
Moschopoulou, G
Manganab, O and
Nomikou, K
Endo, K
Papanastasiou, I
Simonian, A
Περιοδικό:
Biosensors and Bioelectronics
Εκδότης:
ELSEVIER ADVANCED TECHNOLOGY
Λέξεις-κλειδιά:
bioelectric recognition assay (BERA); cell immobilization; cell
proliferation signal; superoxide dismutase; fluorescence microscopy;
oxidative stress
DOI:
10.1016/j.bios.2005.04.022
