Abstract：
:Metastasis is the defining feature of advanced malignancy, yet remains challenging to study in laboratory environments. Here, we describe a high-throughput zebrafish system for comprehensive, in vivo assessment of metastatic biology. First, we generated several stable cell lines from melanomas of transgenic mitfa-BRAF(V600E);p53(-/-) fish. We then transplanted the melanoma cells into the transparent casper strain to enable highly quantitative measurement of the metastatic process at single-cell resolution. Using computational image analysis of the resulting metastases, we generated a metastasis score, μ, that can be applied to quantitative comparison of metastatic capacity between experimental conditions. Furthermore, image analysis also provided estimates of the frequency of metastasis-initiating cells (∼1/120,000 cells). Finally, we determined that the degree of pigmentation is a key feature defining cells with metastatic capability. The small size and rapid generation of progeny combined with superior imaging tools make zebrafish ideal for unbiased high-throughput investigations of cell-intrinsic or microenvironmental modifiers of metastasis. The approaches described here are readily applicable to other tumor types and thus serve to complement studies also employing murine and human cell culture systems.
journal_name
Cancer Resjournal_title
Cancer researchauthors
Heilmann S,Ratnakumar K,Langdon E,Kansler E,Kim I,Campbell NR,Perry E,McMahon A,Kaufman C,van Rooijen E,Lee W,Iacobuzio-Donahue C,Hynes R,Zon L,Xavier J,White RMdoi
10.1158/0008-5472.CAN-14-3319subject
Has Abstractpub_date
2015-10-15 00:00:00pages
4272-4282issue
20eissn
0008-5472issn
1538-7445journal_volume
75pub_type
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