A Simple, Rapid, and Effective Method for Tumor Xenotransplantation Analysis in Transparent Zebrafish Embryos

Monika Verma; Michele Rhodes; Susan Shinton; David L. Wiest

doi:10.3791/66164

Journal

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Cancer Research

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A Simple, Rapid, and Effective Method for Tumor Xenotransplantation Analysis in Transparent Zebrafish Embryos

JoVE Journal
Cancer Research

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JoVE Journal Cancer Research

A Simple, Rapid, and Effective Method for Tumor Xenotransplantation Analysis in Transparent Zebrafish Embryos

1,092 Views

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06:10 min

•

July 12, 2024

DOI:

10.3791/66164-v

DOI:

10.3791/66164-v

•

06:10 min

July 12, 2024

•

4 Views

Monika Verma¹, Michele Rhodes¹, Susan Shinton¹, David L. Wiest¹

¹Nuclear Dynamics and Cancer Program,Fox Chase Cancer Center

Transcript

We seek to gain molecular insight into the control of normal and malignant hematopoiesis and then to exploit those insights to develop new therapies for the treatment of blood cancers. Tumor xenografts are an indispensable tool for evaluating the impact of a genetic alteration or treatment on tumor behavior. However, mice xenografts are expensive, time consuming, and limited in scope.

Using zebrafish for tumor xenotransplantation overcomes all these drawbacks. In this protocol, we demonstrate an easy and rapid mounting process for xenotransplantation, which increases throughput. We further employ cell staining to know the quality of the inoculum and then flow cytometry to assess the tumor progression by evaluating or quantifying the disease burden.

We intend to use this approach to identify novel ways of blocking the function of the Ras/MAP kinase cascade, which drives the vast majority of human cancers. And we’ll do that not by trying to block the catalytic activity of the Ras/MAP kinase cascade but rather by preventing its ability to phosphorylate those cancer-driving substrates it modifies.

Summary

We describe a protocol for xenotransplantation into the yolk of transparent zebrafish embryos that is optimized by a simple, rapid staging method. Post-injection analyses include survival and assessing the disease burden of xenotransplanted cells by flow cytometry.