Basic Colorimetric Proliferation Assays: MTT, WST, and Resazurin

Area of Science:

• Biochemistry
• Cell Biology
• Assay Development

Background:

• Cellular viability and proliferation are critical parameters in cell culture.
• Assessing metabolic activity provides a reliable indicator of cellular health.
• Existing assays require optimization for ease of use and cost-effectiveness.

Purpose of the Study:

• To describe selected biochemical assays for evaluating cellular viability and proliferation.
• To highlight the use of metabolic activity markers, specifically NADH and NADPH, for cell assessment.
• To provide detailed protocols for MTT, WST, and resazurin assays.

Main Methods:

• Measurement of biochemical markers indicative of cellular metabolic activity.
• Utilizing the reducing agents NADH and NADPH as indicators of metabolic state.
• Employing colorimetric changes in specific dyes, detectable by photometry, following reduction by NADH/NADPH.

Main Results:

• MTT, WST, and resazurin assays were selected for their applicability to various cell lines (adherent and suspended).
• These assays are characterized by ease of performance and cost-effectiveness.
• Biochemical reduction of dyes by NADH/NADPH leads to measurable color changes.

Conclusions:

• The described assays (MTT, WST, resazurin) provide robust methods for evaluating cell viability and proliferation.
• These assays are suitable for diverse cell culture applications due to their versatility and economic feasibility.
• Detailed protocols and troubleshooting notes enhance the practical utility of these assays in research settings.