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A quantitative microwell assay for chondrocyte cell adhesion.

R R Miller1, C A McDevitt

  • 1Department of Musculoskeletal Research, Cleveland Clinic Foundation, Ohio 44195.

Analytical Biochemistry
|February 1, 1991
PubMed
Summary

A new assay quantifies live articular chondrocytes using 3-(4,5-dimethylthiazol-2-yl)-2,5-dypenyl tetrazolium bromide (MTT) dye. This method accurately measures chondrocyte attachment to surfaces in microwell plates.

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Area of Science:

  • Biomedical Engineering
  • Cell Biology
  • Biochemistry

Background:

  • Articular chondrocytes are crucial for joint health.
  • Quantifying live chondrocytes in vitro is essential for research.
  • Existing methods may lack sensitivity or specificity.

Purpose of the Study:

  • To establish a reliable assay for measuring live articular chondrocytes.
  • To determine optimal conditions for the assay using MTT dye.
  • To validate the assay's linearity with cell number.

Main Methods:

  • Developed an assay utilizing 3-(4,5-dimethylthiazol-2-yl)-2,5-dypenyl tetrazolium bromide (MTT) dye.
  • Measured absorbance of formazan product generated by live chondrocytes.
  • Optimized MTT concentration (1 mg/ml) and incubation time (3 h).

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  • Utilized microwell plates for cell attachment studies.

    • Main Results:

    • Established a linear relationship between absorbance and live chondrocyte quantity.
    • Optimal MTT concentration and incubation period were identified.
    • The assay successfully quantitated chondrocytes attached to adhesive proteins.

    Conclusions:

    • The MTT-based assay provides a sensitive method for quantifying live articular chondrocytes.
    • This technique is suitable for assessing chondrocyte attachment in research settings.
    • The assay enables accurate measurement of cell viability and attachment on various substrates.