title: Electrochemical sensors, MTT and immunofluorescence assays for monitoring the proliferation effects of cissus populnea extracts on Sertoli cells.
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.
authors.affiliation: Department of Chemistry, Center for Advanced Sensors & Environmental Monitoring, State University of New York at Binghamton, NY 13902-6000, USA.
authors.forename: Elizabeth
authors.initials: E
authors.lastname: Osibote
authors.forename: Naumih
authors.initials: N
authors.lastname: Noah
authors.forename: Omowunmi
authors.initials: O
authors.lastname: Sadik
authors.forename: Dennis
authors.initials: D
authors.lastname: McGee
authors.forename: Modupe
authors.initials: M
authors.lastname: Ogunlesi
mesh_terms: D000818:Animals; D015374:Biosensing Techniques / Q000379:methods*; D049109:Cell Proliferation / Q000187:drug effects*; D002478:Cells, Cultured; D032422:Cissus / Q000737:chemistry*; D004563:Electrochemistry / Q000379:methods; D005303:Fertility Agents, Male / Q000494:pharmacology; D005455:Fluorescent Antibody Technique / Q000379:methods; D008297:Male; D010936:Plant Extracts / Q000494:pharmacology*; D051381:Rats; D012708:Sertoli Cells / Q000187:drug effects* / Q000502:physiology; D013091:Spermatogenesis / Q000187:drug effects / Q000502:physiology; D013778:Tetrazolium Salts / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology
chemical_list: D005303:Fertility Agents, Male; D010936:Plant Extracts; D013778:Tetrazolium Salts; D013844:Thiazoles; C022616:thiazolyl blue