A Clonogenic Assay to Quantify Melanoma Micrometastases in Pulmonary Tissue

Methods Mol Biol. 2021:2265:385-406. doi: 10.1007/978-1-0716-1205-7_28.

Abstract

Metastatic melanoma is one of the most aggressive types of cancers, diffused worldwide and with a significant percentage of lethality. The employment of animal models to test therapeutic strategies against melanoma growth and metastatic spread is of key relevance for cancer biologists. In this regard, the count of metastatic foci in murine lung tissue is one of the recognized methods to monitor macrometastases of melanoma. Here, we illustrate a clonogenic assay method to detect with high sensitivity the presence of single melanoma cells (micrometastases) at the pulmonary level when metastatic foci are still not detectable in the tissue. This method allows for high precision detection and quantification of melanoma metastatic spread to the lung at early stages.

Keywords: B16.F10; Clonogenic assay; Crystal Violet; ImageJ macros; Melanoma; Micrometastasis; Mouse model; R scripts.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Lung Neoplasms / metabolism*
  • Lung Neoplasms / pathology
  • Lung Neoplasms / secondary
  • Melanoma, Experimental / metabolism*
  • Melanoma, Experimental / pathology
  • Mice
  • Neoplasm Metastasis
  • Tumor Stem Cell Assay*