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Jul 1, 2009 (Vol. 29, No. 13)

Upgrading Cell-Based Viability Assays

Novel Approaches More Accurately Reflect the In Vivo Human State

  • Solutions

    Click Image To Enlarge +
    Primary rat hippocampal neurons were cultured and immunolabeled using Millipore assay components. Neurons are labeled with beta-III-tubulin (green), and presynaptic vesicles are labeled with synaptophysin (red).

    “We approach cell-based assays on two fundamental levels—providing solutions for characterizing cellular events across a wide range of cell types and using cells as biological test tubes to engineer specific assays for research and drug discovery,” says Jeff Till, Ph.D., marketing director for drug discovery at Millipore. The company has introduced kits for studying cellular toxicity using high-content image analysis.

    “We cover a range of cellular toxicity responses such as DNA damage and oxidative stress,” Dr. Till says. A number of new products focus on assessment of the effects of drugs on the nervous system. These include a screening kit composed of high-quality, validated, target-specific detection reagents for profiling neurotoxicity, neurite outgrowth, and neuronal morphology in a wide variety of mammalian cell types.

    Additional cell-based assay products available from Millipore include cell-signaling kits designed around a variety of cell lines engineered for drug discovery through the use of specific reporter systems. Noteworthy are mouse and human stem cell lines for the investigation of compounds that affect cellular differentiation.

  • Gene Manipulation in Living Cells

    Sigma-Aldrich and Sangamo Biosciences have partnered to exploit an interventional technology in genome science for the development of new material for cell-based assays.

    “Using a zinc finger nuclease-based approach we are now able to engineer the genome of living cells,” states David Smoller, Ph.D., president of the research biotechnology business unit at Sigma-Aldrich.

    The CompoZr™ zinc finger nuclease (ZFN) technology for targeted genome editing allows the insertion, deletion, or alteration of targeted sequences, including the placement of a reporter gene in front or at the 3´ end of any endogenous locus that the investigator may choose.

    The technology is modular—the zinc finger domain targets to a specific sequence and the heterodimeric nuclease cleaves the genomic DNA, creating a double-stranded break. This break in the DNA stimulates the natural repair mechanisms of the cell, allowing an exogenous transfected DNA sequence to be integrated into the region of interest in the genome.

    The CompoZr ZFN system allows the investigator to engineer a wide range of possibilities for custom cell lines to be used in cell-based assays. The procedure is fast and straightforward, and because it can be performed on living cells, it would lend itself especially to redesigning embryos, including Drosophila, zebrafish, mice, and other important molecular model systems.

Readers' Comments

Posted 07/10/2009 by Sr. Director, Business Development

Very insightful article that highlights multiple new technologies being advanced. One concern that I do not see being addressed is the drive to make our in vitro assays more physiologically relevant by removing and replacing non-human components when possible. Cells do b ehave differently when cultured on plastic or Matrigel when compared to fully human ECMs. There are dramatic morphological as well as molecular gene expression differences that lead to aberrant and misleading results. Minimizing these impacts on in vitro assays and maximizing human physiological relevance is the future of cell-based assays.

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