Human melanoma cells transplanted into zebrafish proliferate, migrate, produce melanin, form masses and stimulate angiogenesis in zebrafish
Human melanoma cells transplanted into zebrafish proliferate, migrate, produce melanin, form masses and stimulate angiogenesis in zebrafish
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DOI：
10.1007/s10456-006-9040-2
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发表时间：
2006-01-01
期刊：
影响因子：
9.8
通讯作者：
McGrath, Patricia
中科院分区：
文献类型：
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作者：
Haldi, Maryann;Ton, Christopher;McGrath, Patricia
在这项研究中，我们优化了异种移植WM-266-4，转移性黑色素瘤细胞系，包括斑马鱼的位置和阶段的移植，细胞数量，注射方法，和斑马鱼孵育温度的参数。黑色素瘤细胞在体内增殖、迁移并形成团块。我们移植了另外两种癌细胞系，结肠直肠癌细胞系SW 620和胰腺癌细胞系FG CAS/Crk，这些人类癌症也在斑马鱼中形成肿块。我们还移植了CCD-1092 Sk，一种从正常包皮建立的人成纤维细胞系，该细胞系迁移，但不增殖或形成肿块。我们通过解离异种移植的斑马鱼，将来自每个斑马鱼的CM-DiI标记的人细胞的等分试样分配到血细胞计数器载玻片上，然后目视计数荧光标记的癌细胞的数量，来定量增殖的黑色素瘤和正常皮肤成纤维细胞的数量。由于斑马鱼是透明的，直到约30 dpf，标记的黑色素瘤细胞和斑马鱼内皮细胞（EC）的相互作用，可以可视化的整体安装免疫化学染色。Phy-V，小鼠抗斑马鱼单克隆抗体（mAb），特异性标记激活EC和血管母细胞染色后，使用免疫组织学和2-光子显微镜，我们观察到激活斑马鱼EC嵌入人黑色素瘤细胞团。斑马鱼模型提供了一种快速有效的方法来评估肿瘤发生各个阶段的人类癌细胞。
In this research, we optimized parameters for xenotransplanting WM-266-4, a metastatic melanoma cell line, including zebrafish site and stage for transplantation, number of cells, injection method, and zebrafish incubation temperature. Melanoma cells proliferated, migrated and formed masses in vivo. We transplanted two additional cancer cell lines, SW620, a colorectal cancer cell line, and FG CAS/Crk, a pancreatic cancer cell line and these human cancers also formed masses in zebrafish. We also transplanted CCD-1092Sk, a human fibroblast cell line established from normal foreskin and this cell line migrated, but did not proliferate or form masses. We quantified the number of proliferating melanoma and normal skin fibroblasts by dissociating xenotransplant zebrafish, dispensing an aliquot of CM-DiI labeled human cells from each zebrafish onto a hemocytometer slide and then visually counting the number of fluorescently labeled cancer cells. Since zebrafish are transparent until approximately 30 dpf, the interaction of labeled melanoma cells and zebrafish endothelial cells (EC) can be visualized by whole-mount immunochemical staining. After staining with Phy-V, a mouse anti-zebrafish monoclonal antibody (mAb) that specifically labels activated EC and angioblasts, using immunohistology and 2-photon microscopy, we observed activated zebrafish EC embedded in human melanoma cell masses. The zebrafish model offers a rapid efficient approach for assessing human cancer cells at various stages of tumorigenesis.