Methods in cancer research: Assessing therapy response of spheroid cultures by life cell imaging using a...

Area of Science:

• Biomedical Engineering
• Cancer Research
• Cell Biology

Background:

• Three-dimensional (3D) spheroid cultures are more clinically relevant than 2D models for predicting cancer therapy response.
• Existing live-dead staining methods for spheroids are often costly, toxic, or lack long-term stability.
• Accurate monitoring of therapy response in 3D cancer models is crucial for drug development.

Purpose of the Study:

• To develop a cost-effective and stable live-dead staining protocol for monitoring cancer spheroid therapy response.
• To establish a more accurate method for assessing treatment efficacy in 3D cancer models.
• To validate the protocol using glioblastoma spheroid models.

Main Methods:

• Utilized calcein-AM (live cells) and Helix NP™ Blue (dead cells) for live-dead staining in spheroid cultures.
• Employed ICY BioImage Analysis and Z-stacks projection for quantitative viability assessment.
• Visualized spheroid cellular architecture using confocal microscopy.

Main Results:

• The developed method enabled stable monitoring of spheroid therapy response for up to 10 days.
• Quantitative viability analysis via image processing proved more accurate than spheroid size measurements.
• Glioblastoma spheroids showed reduced viability after 7-day treatment with temozolomide.

Conclusions:

• The novel calcein-AM and Helix NP™ Blue staining method provides a cost-effective, stable, and accurate approach for assessing therapy response in cancer spheroids.
• This versatile method can be adapted for various cancer types and 3D culture models.
• The technique enhances the predictive value of 3D cancer models for therapeutic efficacy evaluation.