Wth phase, metastatic growth phase) retained their graded invasive qualities in

Wth phase, metastatic growth phase) retained their graded invasive qualities in vivo. For the in vivo experiment we chose the rhombencephalon as transplantation site. However, the method itself was not described in detail.Non-transformed Primary Human Melanocytes do not Perform Neural Crest Cell MigrationTo demonstrate that the neural crest migration spontaneously performed by melanoma cells upon transplantation into the neural tube was due to re-expression of embryonic traits, aggregates from primary human melanocytes were also injected into the neural tube. Histological evaluation demonstrated that the melanocyte aggregates integrated into the roof plate but did not perform neural crest migration (Figures 3D ), indicating that the neural crest-derived melanocytes residing in the differentiated stratified epithelium of the skin have lost the capability of spontaneous neural crest migration.Pre-treatment with the TGFbeta Family Members BMP-2 or Nodal Induces Invasive Migration of Human Melanocytes in 18325633 the Optic CupWe next asked whether the results gained on human melanocytes in the neural tube of the chick embryo (compare Figures 3D ) could be re-produced in an 298690-60-5 biological activity ectopic site. We therefore injected the melanocytes into the optic cup as described above. Since in our previous reports we saw a BMP-dependence of neural crest migration of melanoma cells [16] and of invasive migration of 26001275 melanoma cells in the optic cup [17], we now asked whether BMP-2 or nodal (both TGF-beta family members) could drive invasive migration in the melanocytes. As expected, untreated melanocytes formed loosely aggregated tumors behind the lens, adjacent to the hyaloid vessels and in the developing vitreous body. The human melanocytes were identified in the chick embryo by their specific pigmentation and morphology. The untreated melanocytes showed no invasion (Fig. 4, upper row). In the melanocytes pre-treated with BMP-2 or nodal we also observed the formation of loosely aggregated tumors in similar locations. In contrast to untreated melanocytes, single and groups of BMP-2 pre-treated melanocytes could be found in the lens epithelium, the retina, in the hyaloid vessels, and, most pronounced, invading the choroid (Fig. 4, middle row). In the group of nodal pre-treated melanocytes single melanocytes invading the choroid and the hyaloid vessels were observed (Fig. 4, lower row). For all three experimental groups for evaluation we grouped the melanocytes according to the compartments in which they were found: injection channel, choroid, hyaloid vessels, vitreous body, and behind the lens (compare Table 1).Transplantation into the Optic Cup is a Model for Invasive Migration of Melanoma CellsAs second niche for the investigation of invasion, the embryonic optic cup was chosen. Upon transplantation of B16-F1 melanoma cells into stage 19?0 HH embryos and Rubusoside site incubation for 72 h, histological evaluation illustrated that one part of the melanoma cells had remained behind the lens at the spot of transplantation, while a second part had formed tumors in and invaded the choroid (highly vascularized, loose mesenchymal connective tissue) in the region of the prospective anterior eye chamber (Figure 3G ). In some embryos, the cells had destroyed the lens and invaded the hyaloid vessels (not shown). Malignant growth of melanoma cells in the embryonic optic cup is also enhanced by BMP-2 and can be blocked by noggin [17].Transplantation into the Brain Vesicles is a Model for Brain Metast.Wth phase, metastatic growth phase) retained their graded invasive qualities in vivo. For the in vivo experiment we chose the rhombencephalon as transplantation site. However, the method itself was not described in detail.Non-transformed Primary Human Melanocytes do not Perform Neural Crest Cell MigrationTo demonstrate that the neural crest migration spontaneously performed by melanoma cells upon transplantation into the neural tube was due to re-expression of embryonic traits, aggregates from primary human melanocytes were also injected into the neural tube. Histological evaluation demonstrated that the melanocyte aggregates integrated into the roof plate but did not perform neural crest migration (Figures 3D ), indicating that the neural crest-derived melanocytes residing in the differentiated stratified epithelium of the skin have lost the capability of spontaneous neural crest migration.Pre-treatment with the TGFbeta Family Members BMP-2 or Nodal Induces Invasive Migration of Human Melanocytes in 18325633 the Optic CupWe next asked whether the results gained on human melanocytes in the neural tube of the chick embryo (compare Figures 3D ) could be re-produced in an ectopic site. We therefore injected the melanocytes into the optic cup as described above. Since in our previous reports we saw a BMP-dependence of neural crest migration of melanoma cells [16] and of invasive migration of 26001275 melanoma cells in the optic cup [17], we now asked whether BMP-2 or nodal (both TGF-beta family members) could drive invasive migration in the melanocytes. As expected, untreated melanocytes formed loosely aggregated tumors behind the lens, adjacent to the hyaloid vessels and in the developing vitreous body. The human melanocytes were identified in the chick embryo by their specific pigmentation and morphology. The untreated melanocytes showed no invasion (Fig. 4, upper row). In the melanocytes pre-treated with BMP-2 or nodal we also observed the formation of loosely aggregated tumors in similar locations. In contrast to untreated melanocytes, single and groups of BMP-2 pre-treated melanocytes could be found in the lens epithelium, the retina, in the hyaloid vessels, and, most pronounced, invading the choroid (Fig. 4, middle row). In the group of nodal pre-treated melanocytes single melanocytes invading the choroid and the hyaloid vessels were observed (Fig. 4, lower row). For all three experimental groups for evaluation we grouped the melanocytes according to the compartments in which they were found: injection channel, choroid, hyaloid vessels, vitreous body, and behind the lens (compare Table 1).Transplantation into the Optic Cup is a Model for Invasive Migration of Melanoma CellsAs second niche for the investigation of invasion, the embryonic optic cup was chosen. Upon transplantation of B16-F1 melanoma cells into stage 19?0 HH embryos and incubation for 72 h, histological evaluation illustrated that one part of the melanoma cells had remained behind the lens at the spot of transplantation, while a second part had formed tumors in and invaded the choroid (highly vascularized, loose mesenchymal connective tissue) in the region of the prospective anterior eye chamber (Figure 3G ). In some embryos, the cells had destroyed the lens and invaded the hyaloid vessels (not shown). Malignant growth of melanoma cells in the embryonic optic cup is also enhanced by BMP-2 and can be blocked by noggin [17].Transplantation into the Brain Vesicles is a Model for Brain Metast.