Abstract
BackgroundWe describe the development of an electrochemical sensor array for monitoring the proliferation effects of cissus populnea plant extracts on TM4 Sertoli cells.MethodsThe 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.ResultsThis 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.ConclusionsOverall, 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.
Highlights
We describe the development of an electrochemical sensor array for monitoring the proliferation effects of cissus populnea plant extracts on TM4 Sertoli cells
The immediate objective was to compare the performance of three analytical techniques on the proliferation effects: electrochemical sensor arrays (DOX-96), colorimetric assay using MTT and fluorescence techniques
Spermatogenesis and proliferation effect of cells Follicle stimulating hormone (FSH) supports the growth of Sertoli cells and abnormal follicle stimulating hormones (FSH) levels in both male and female can be related to infertility
Summary
We describe the development of an electrochemical sensor array for monitoring the proliferation effects of cissus populnea plant extracts on TM4 Sertoli cells. Cissus populnea is one of the several climbing tropical shrubs that are believed to promote fertility in males and females, the mechanism is unclear [1]. Another member of the family (Cissus sicyoides) has been reported for the treatment of rheumatism, epilepsy and stroke [2]. Extracts from the plant have been screened for antimicrobial activities [3], treatment of Extracts from the stem of Cissus populnea are believed to improve fertility in men with low sperm count [1]. It is believed that the hormonal regulation of spermatogenesis is mediated by the Sertoli cells since they either partially or completely surround every germ cell [11]
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