Electrochemical sensors, MTT and immunofluorescence assays for monitoring the proliferation effects of cissus populnea extracts on Sertoli cells
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Date
2011
Authors
Osibote, E.
Noah, N.
Sadik, O.
McGee, D.
Ogunlesi, M.
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Abstract
Background: We describe the development of an electrochemical sensor array for monitoring the proliferation
effects of cissus populnea plant extracts on TM4 Sertoli cells.
Methods: The proliferation activities of the extracts on Sertoli cells were studied using a high-throughput
electrochemical sensor array (DOX-96) and the analytical sensor characteristics were compared with conventional
colorimetric MTT (3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide) assay and fluorescence spectroscopy.
Results: This work shows that there is a definite positive trend in the proliferation effect of the extract of Cissus
populnea on the TM4 Sertoli cells. All of the three techniques confirmed that the most effective concentration for
the proliferation is 10 ppm. At this concentration, the proliferation effect was established around 120% for both
DOX-96 and MTT techniques, whereas fluorescence assays showed a higher level (120-150%). DOX-96 showed a
lower limit of detection (1.25 × 10(4) cells/ml); whereas the LOD recorded for both MTT and fluorescence
techniques was 2.5 × 10(4) cells/ml. Visual examination of the cells by means of confocal fluorescence microscopy
confirmed the proliferation of Sertoli cells as was determined using the MTT assay. This investigation provides a
confident interpretation of the results and proved that the most effective concentration for the proliferation using
Cissus populnea plant extract is 10 ppm.
Conclusions: Overall, the DOX results compared well with the conventional methods of checking proliferation of
cells. The fascinating feature of the sensor array is the ability to provide continuous proliferation experiments with
no additional reagents including 96 simultaneous electrochemical experiments. The use of the DOX-96 could
reduce a typical bioassay time by 20-fold. Thus the DOX-96 can be used as both a research tool and for practical
cell culture monitoring.
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Keywords
Electrochemical sensor array , Plant extracts , Sertoli cells , Fluorescence assays , Research Subject Categories::NATURAL SCIENCES::Chemistry
Citation
Osibote, E., [Et..al] (2011). Electrochemical sensors, MTT and immunofluorescence assays for monitoring the proliferation effects of cissus populnea extracts on Sertoli cells. Reproductive Biology and Endocrinology, 9(1), 65.