Journal: PLoS ONE

Article Title: The Chick Embryo as an Experimental System for Melanoma Cell Invasion

doi: 10.1371/journal.pone.0053970

Figure Lengend Snippet: (A) Schematic drawing depicting ventral and medial neural crest migration pathways. n.c. neural crest; n.t. neural tube; s.t. sympathetic trunk. (B) Chick embryo 24 h after transplantation of SKMel28 melanoma cells into the neural tube. Melanoma cells (visualized by HMB45 immunoreactivity) spontaneously resuming neural crest migration have a stretched, mesenchymal-like morphology (arrows). (C) At the site of destination along the ventral migration pathway (para-aortic sympathetic ganglia) melanoma cells undergo apoptosis, visualized by TUNEL staining. (D,E) Chick embryo 24 h after transplantation of benign primary human melanocytes into the neural tube. Melanocytes (showing a compact, epithelial-like morphology) are encountered only in the lumen of the neural tube and, in part, integrated into the roof plate with no neural crest migration. (F) Melan A immunoreactivity confirms the melanocytic origin of the cells. (G) Schematic drawing of chick embryo 72 h after transplantation of B16-F1 melanoma cells into the optic cup. (H) Histological correlate of schematic drawing. Already in H&E staining the transplanted, invasively migrating melanoma cells are visible (arrows). (I) Single melanoma cells (identified by HMB45 immunoreactivity) form a tumor, and single melanoma cells invade the choroid of the optic cup (arrows). (J) Chick embryo 96 h after transplantation of human metastatic melanoma cells into the brain vesicle at the hindbrain (rhombencephalon). The cells form a large tumor in the dorsal neuroepithelium with (K) single HMB45 positive cells infiltrating the surrounding brain tissues. (L) MIB1 immunohistochemistry (proliferation marker not cross-reacting with chick cells) identifies melanoma cells during haematogenous spreading in blood vessels among host erythrocytes and lymphocytes, and in the surrounding neural tissue.

Article Snippet: For each series of transplantation, one of the following cells were used as aggregates or cell suspensions: Mouse B16-F1 metastatic melanoma cells (gifted from ); human SKMel28 metastatic melanoma cells (purchased as part of the NCI60 panel of cancer cells from the NCI); human 451LU metastatic melanoma cells (gifted from Meenhard Herlyn, Wistar Institute, Philadelphia, USA ), or human melanocytes (human epidermal melanocytes neonatal (HEMn), CellSystems, Troisdorf, Germany, cultivated in Lifeline's DermaLife M medium (CellSystems)).

Techniques: Migration, Transplantation Assay, TUNEL Assay, Staining, Immunohistochemistry, Marker

Journal: PLoS ONE

Article Title: The Chick Embryo as an Experimental System for Melanoma Cell Invasion

doi: 10.1371/journal.pone.0053970

Figure Lengend Snippet: Untreated, BMP-2 or nodal pre-treated melanocytes were injected into the optic cup of the chick embryo (stage 20 HH). After 72 h of further incubation, the embryos were analyzed for tumor growth and invasion. Untreated melanocytes formed loosely aggregated tumors adjacent to the hyaloid vessels (left image in upper row), in the developing vitreous body and behind the lens (right image in upper row) without invasion. The BMP-2 and nodal groups formed tumors in similar locations. In the BMP-2 group single melanocytes invaded the lens epithelium (insert in left image in middle row), the retina, the hyaloid vessels, and the choroid (right image in middle row; arrows pointing at melanocytes). In the nodal group single melanocytes invaded the choroid (lower row, arrows in right image) and the hyaloid vessels.

Article Snippet: For each series of transplantation, one of the following cells were used as aggregates or cell suspensions: Mouse B16-F1 metastatic melanoma cells (gifted from ); human SKMel28 metastatic melanoma cells (purchased as part of the NCI60 panel of cancer cells from the NCI); human 451LU metastatic melanoma cells (gifted from Meenhard Herlyn, Wistar Institute, Philadelphia, USA ), or human melanocytes (human epidermal melanocytes neonatal (HEMn), CellSystems, Troisdorf, Germany, cultivated in Lifeline's DermaLife M medium (CellSystems)).

Techniques: Injection, Incubation