Optimization of fluorescent cell-based assays for high-throughput analysis using microchamber array chip formats

Abstract This study describes the optimization of fluorescent cell-based assays using microchamber array chip formats as well as using automatic nanoliter volumes of sample dispensing system for high-throughput screening analysis of anticancer drugs. Cell-based assays can be employed efficiently in the screening of potential anticancer drug candidates and bioactive compounds with distinct biological function. Identification and development of cell-based assays adapted to high-throughput screening requirements is important when screening chemicals for their potential anticancer properties. Cell-based screening assays using microchamber array chip formats and automatic nanoliter volumes of sample dispensing system requires an optimization as a prerequisite for parameters including assay liquid volume and number of cells per each chamber, and the total cell-based assay itself. Further, the anticancer effect of mitomycin C was studied as an example against human cervical carcinoma cell line-HeLa 229 using cell-based assay that was optimized on chamber array chip formats and determined the cytotoxicity of mitomycin C by measuring the cell proliferation of HeLa with Calcein-AM fluorescent dye. The cell-based screening assay that was performed using chamber array chip formats was compared with the conventional 96-well plate formats was discussed. The assay described in this study is rapid, simple and inexpensive that is desirable in selecting anticancer candidates.

[1]  Rose Microdispensing technologies in drug discovery. , 1999, Drug discovery today.

[2]  Yasutaka Morita,et al.  Application of a microchamber array for DNA amplification using a novel dispensing method. , 2002, Archives of histology and cytology.

[3]  S. Lautraite,et al.  Optimisation of cell-based assays for medium throughput screening of oxidative stress. , 2003, Toxicology in vitro : an international journal published in association with BIBRA.

[4]  A. Klippel,et al.  Unravelling novel intracellular pathways in cell-based assays. , 2002, Drug discovery today.

[5]  S A Sundberg,et al.  High-throughput and ultra-high-throughput screening: solution- and cell-based approaches. , 2000, Current opinion in biotechnology.

[6]  T. Sakaeda,et al.  Cytotoxic effects of 27 anticancer drugs in HeLa and MDR1-overexpressing derivative cell lines. , 2002, Biological & pharmaceutical bulletin.

[7]  M Webb,et al.  Ultra-High Throughput Screen of Two-Million-Member Combinatorial Compound Collection in a Miniaturized, 1536-Well Assay Format , 2000, Journal of biomolecular screening.

[8]  R. Schlegel,et al.  Presence and expression of human papillomavirus sequences in human cervical carcinoma cell lines. , 1985, The American journal of pathology.

[9]  J. Wölcke,et al.  Miniaturized HTS technologies - uHTS. , 2001, Drug discovery today.

[10]  Thomas D.Y. Chung,et al.  Assay Miniaturization for Ultra-High Throughput Screening of Combinatorial and Discrete Compound Libraries: A 9600-Well (0.2 Microliter) Assay System , 1998 .

[11]  R. Hertzberg,et al.  High-throughput screening: new technology for the 21st century. , 2000, Current opinion in chemical biology.