Protocol for quantifying xenografted human cancer cells in zebrafish larvae using Cellpose.

Area of Science:

• Cancer Biology
• Zebrafish Models
• Biotechnology

Background:

• Glioblastoma stem cells (GSCs) are crucial for tumor growth and recurrence.
• Accurate quantification of GSCs in vivo is essential for developing effective therapies.
• Existing methods for GSC quantification can be labor-intensive and subjective.

Purpose of the Study:

• To present a detailed protocol for quantifying xenografted human glioblastoma stem cells (GSCs) in zebrafish larvae.
• To establish a semi-automatic method for GSC segmentation and quantification.
• To enable unbiased assessment of GSC survival and proliferation in xenograft models.

Main Methods:

• Orthotopic xenotransplantation of GSCs into the midbrain of zebrafish larvae.
• Immunofluorescent labeling and confocal imaging of xenografted cells.
• GSC quantification utilizing the Cellpose algorithm for semi-automatic segmentation.

Main Results:

• A robust protocol for GSC xenotransplantation and imaging in zebrafish.
• Successful application of the Cellpose algorithm for accurate GSC segmentation.
• Demonstration of unbiased quantification of GSC survival and proliferation.

Conclusions:

• The presented protocol offers a reliable technique for quantifying human cancer cells in zebrafish xenografts.
• This method facilitates objective assessment of therapeutic efficacy targeting GSCs.
• The Cellpose-based approach enhances the efficiency and accuracy of GSC research.