Experimental mice for the development of an orthotopic model of bone malignancies.
Britta VormoorHenrike K. KniziaMichael A. BateyGilberto S. AlmeidaIan WilsonPetra DildeyAbhishek SharmaHelen BlairI. Geoff HideOlaf HeidenreichJosef VormoorRoss J. MaxwellChris M. Bacon
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In recent decades, subcutaneously implanted patient-derived xenograft tumors or cultured human cell lines have been increasingly recognized as more representative models to study human cancers in immunodeficient mice than traditional established human cell lines in vitro. Recently, orthotopically implanted patient-derived tumor xenograft (PDX) models in mice have been developed to better replicate features of patient tumors. A liver orthotopic xenograft mouse model is expected to be a useful cancer research platform, providing insights into tumor biology and drug therapy. However, liver orthotopic tumor implantation is generally complicated. Here we describe our protocols for the orthotopic implantation of patient-derived liver-metastatic uveal melanoma tumors. We cultured human liver metastatic uveal melanoma cell lines into immunodeficient mice. The protocols can result in consistently high technical success rates using either a surgical orthotopic implantation technique with chunks of patient-derived uveal melanoma tumor or a needle injection technique with cultured human cell line. We also describe protocols for CT scanning to detect interior liver tumors and for re-implantation techniques using cryopreserved tumors to achieve re-engraftment. Together, these protocols provide a better platform for liver orthotopic tumor mouse models of liver metastatic uveal melanoma in translational research.
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Orthotopic liver transplantation
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Background: Gallbladder carcinoma (GBC) is the most common carcinoma of the biliary system. Among its research models, orthotopic xenograft models, important research tools, have been rarely reported in the literature however. Aim: To explore establishment of an orthotopic xenograft model and to evaluate the advantage and disadvantage as compared with other models. Materials and Methods: Subcutaneous xenograft and orthotopic xenograft models of gallbladder carcinoma in nude mice were established and compared with human gallbladder carcinomas. Results: For the orthotopic xenograft model and clinical gallbladder carcinomas, the lymph node metastatic rates were 69.2% and 53.3% (p>0.05); ascites generation rates, 38.5% and 11.7%(p<0.05); liver invasive rates, 100% and 61.7%(p<0.05); and lymphatic vessel densities (LVD), $10.4{\pm}3.02$ and $8.77{\pm}2.92$ (p>0.05), respectively. In the subcutaneous xenograft model, no evidence of ascites generation, lymph node metastasis and liver metastasis were found, and its LVD was lower ($4.56{\pm}1.53$ , p<0.05). Conclusions: Compared with the subcutaneous xenograft model, the orthotopic xenograft model better simulates clinical gallbladder carcinoma in terms of metastasis and invasion, which may be attributed to the difference in microenvironment and LVD.
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以细胞因子白细胞介素(IL)-12为基础的抗肿瘤免疫疗法有望成为治疗肝脏肿瘤的有效手段之一.以往的IL-12抗肿瘤基因治疗研究中多采用皮下接种肝癌模型,肿瘤的生长环境与临床实际情况相差甚远。
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AbstractA number of xenograft models have been developed for human lung cancer. These include subcutaneous (sc)-implant models and implantation under the renal capsule, but these models have not been sufficiently representative of the clinical situation (1). The studies of McLemore et al. (2,3) have utilized the orthotopic concept to develop more relevant lung-tumor models in nude mice. The first model developed by McLemore et al. (2) utilized the growth of human lung cancer cell lines in the bronchioloalveolar region of the right lung of nude mice implanted via an intrabronchial injection. Suspensions of disaggregated fresh tumor specimens were also implanted intrabronchially (ib) by this group. These tumors grew intrabronchially much more extensively than the same tumors inoculated sc. However, most of the tumors propagated ib were localized to the right lung, with only 1% metastasizing to the left lung, 2% to the trachea, 6% to the peritracheal area, and only 3% spreading distantly to lymph nodes, liver, or spleen. McLemore et al. (3) subsequently developed a second model by injecting lung tumor cells via an intrathoracic route into the pleural space. This model seems similar to the intrabronchial model in that extensive local growth occurs with little metastatic spread. KeywordsGreen Fluorescent ProteinNude MouseEnhance Green Fluorescent ProteinLeft LungGreen Fluorescent Protein ExpressionThese keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.
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