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Functional Assays and Toxicity Screening

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Tissue Engineering

Part of the book series: Learning Materials in Biosciences ((LMB))

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Abstract

This chapter covers the most commonly used cell viability assays. These assays are needed for students to properly evaluate the survival of the cells and/or tissue constructs during individual steps of the tissue engineering process. Different viability assays rely on different aspects of cell metabolism with various factors possibly impacting the outcome of the assay. Therefore, a proper interpretation of results requires understanding the mechanisms of each assay. Here we briefly discuss DNA-dye binding methods, lactate dehydrogenase release, resazurin and tetrazolium conversions, and the use of LIVE/DEAD stains as means to evaluate cell viability. We also explain how to include three types of controls, the importance of working within the linear part of the standard curve, and the need to have triplicates to ensure the reproducibility of the measurements.

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References and Further Reading

  1. T.L. Riss, R.A Moravec, A.L. Niles et al. Cell Viability Assays. 2016. In: Sittampalam GS, Grossman A, Brimacombe K, et al., editors. Assay Guidance Manual [Internet]. Bethesda (MD): Eli Lilly & Company and the National Center for Advancing Translational Sciences, https://www.ncbi.nlm.nih.gov/books/NBK144065; Präbst K, Engelhardt H, Ringgeler S, Hübner H. Basic Colorimetric Proliferation Assays: MTT, WST, and Resazurin. Methods Mol Biol. (1601), 1–17 (2017)

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Author information

Authors and Affiliations

  1. L. Orbeli Institute of Physiology, Yerevan, Armenia

    Hovhannes Arestakesyan

  2. The George Washington University, Washington, DC, USA

    Narine Sarvazyan

Authors
  1. Hovhannes Arestakesyan
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  2. Narine Sarvazyan
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Corresponding author

Correspondence to Narine Sarvazyan .

Editor information

Editors and Affiliations

  1. The George Washington University, Washington, DC, USA

    Narine Sarvazyan

Self-Check Questions

Self-Check Questions

  1. Q.5.1.

    Which of the following assays is considered to be a tissue-specific functional assay?

    1. A.

      Release of lactate dehydrogenase

    2. B.

      Permeability to Trypan blue

    3. C.

      Chondroitin production

    4. D.

      Reduction of resazurin

  1. Q.5.2.

    Which of the following assays is often used to noninvasively evaluate the survival of implanted tissue constructs?

    1. A.

      Tensile strength

    2. B.

      Load-bearing

    3. C.

      Bioluminescence

    4. D.

      LIVE/DEAD assay

  1. Q.5.3.

    What type of control is NOT included when testing response to a drug or a treatment?

    1. A.

      Blank

    2. B.

      Neutral control

    3. C.

      Positive control

    4. D.

      Negative control

  1. Q.5.4.

    Which assay recommends the transfer of accumulated media to a new well for viability testing?

    1. A.

      Resazurin

    2. B.

      LDH

    3. C.

      MTT

    4. D.

      Ethidium bromide

  1. Q.5.5.

    Viability assays can rely on different aspects of cell metabolism, EXCEPT

    1. A.

      Mitochondrial activity

    2. B.

      Protein turnover

    3. C.

      ATP production

    4. D.

      DNA mutation rate

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Arestakesyan, H., Sarvazyan, N. (2020). Functional Assays and Toxicity Screening. In: Sarvazyan, N. (eds) Tissue Engineering. Learning Materials in Biosciences. Springer, Cham. https://doi.org/10.1007/978-3-030-39698-5_5

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