| In vivo Efficacy Studies in Cell Line and Patient-derived Xenograft Mouse Models
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Author:
Date:
2017-01-05
[Abstract] In vivo xenograft models derived from human cancer cells have been a gold standard for evaluating the genetic drivers of cancer and are valuable preclinical models for evaluating the efficacy of cancer therapeutics. Recently, patient-derived tumorgrafts from multiple tumor types have been developed and shown to more accurately recapitulate the molecular and histological heterogeneity of cancer. Here we detail the procedures for developing patient-derived xenograft models from breast cancer tissue, cell-based xenograft models, serial tumor transplantation, tumor measurement, and drug treatment.
[摘要] 衍生自人类癌细胞的体内异种移植模型已经成为评估癌症遗传学驱动因素的黄金标准，并且是用于评估癌症治疗药物功效的有价值的临床前模型。最近，已经开发了来自多种肿瘤类型的患者衍生的移植物，并且显示出更准确地概括了癌症的分子和组织学异质性。在这里，我们详细介绍了从乳腺癌组织，基于细胞的异种移植模型，连续肿瘤移植，肿瘤测量和药物治疗开发患者衍生异种移植模型的程序。
背景 异种移植模型已经成为研究癌症基因驱动因素和确定癌症治疗药物的潜在功效的有力方法。发现T细胞缺乏的无胸腺裸鼠（nu / nu）小鼠和T细胞和B细胞缺陷型严重联合免疫缺陷（scid / scid）小鼠（Flanagan， 1966; Bosma和Carroll，1991）。由于这些发现，其他免疫受损的小鼠模型已经变得可用，包括重组激活基因2（Rag2）敲除小鼠，非肥胖（NOD） - 小鼠和NOD-scid < il2rgamma（null）小鼠（也称为nsg小鼠）（shinkai等人，1992;="" prochazka等人，1992;="">
＆nbsp;在开发包括注射或植入部位的异种移植模型时，应考虑几个选项。由于生长测量和成像的肿瘤可及性，皮下异种移植物通常用于体内研究;然而，该模型的显着限制是大多数癌细胞缺乏正常的基质微环境。原位异种移植物提供互补的基质微环境;然而，根据原位点，包括更复杂的外科手术，测量肿瘤生长或反应的难度以及啮齿动物基质的限制（Talmadge等人，2007），该途径也存在缺点。许多癌症研究尤其是乳腺癌研究中广泛应用原位异种移植物。注入乳腺脂肪垫是一个相对简单的程序，允许乳腺癌细胞的可见和可测量的生长。尽管乳腺脂肪垫为乳腺癌细胞提供了互补的组织部位，但重要的是注意到人和啮齿动物乳腺基质和激素环境之间存在明显差异。 ...
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| In vitro Differentiation of Murine Innate Lymphoid Cells from Common Lymphoid Progenitor Cells
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Author:
Date:
2016-03-20
[Abstract] Subtypes of innate lymphoid cells (ILC), defined based on their cytokine secretion profiles and transcription factor expression, are important for host protection from pathogens and maintaining tissue homeostasis. ILCs develop from common lymphoid progenitors (CLP) in the bone marrow. Using the methods described here, we have previously shown that loss of the transcriptional regulator TOX (Thymocyte-selection associated HMG-box protein) leads to specific changes in ILC development and differentiation. Here, we describe how to obtain ILCs from in vivo isolated CLP grown in vitro.
[摘要] 先天淋巴细胞（ILC）的亚型，基于其细胞因子分泌谱和转录因子表达来定义，对于宿主保护免于病原体和维持组织内稳态是重要的。 ILC从骨髓中的普通淋巴祖细胞（CLP）发展而来。 使用本文所述的方法，我们以前显示转录调节器TOX（胸腺细胞选择相关的HMG框蛋白）的损失导致ILC发展和分化的具体变化。 在这里，我们描述了如何从体内分离的CLP获得ILC。
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| Agrobacterium rhizogenes-Based Transformation of Soybean Roots to Form Composite Plants
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Author:
Date:
2016-01-20
[Abstract] Transgenic soybean roots of composite plants are a powerful tool to rapidly test the function of genes and activity of gene promoters. No tissue culture is needed, thus avoiding loss of valuable material due to contamination. This is a simple technique that requires less training and care than tissue culture techniques. Furthermore, it takes less time to produce transgenic roots than techniques using sterile tissue culture. If the transgenic roots are to be challenged with a pathogen, there is no need to produce axenic pathogens with this technique, because sterile tissue culture medium is not used. Therefore, there is no agar medium on which contaminants may grow resulting in obscured results or diseased roots. Here, we describe the production of transgenic soybean roots on 7-day-old ...
[摘要] 复合植物的转基因大豆根是快速测试基因功能和基因启动子活性的有力工具。不需要组织培养，因此避免了由于污染导致的有价值材料的损失。这是一种简单的技术，比组织培养技术需要更少的培训和护理。此外，产生转基因根的时间比使用无菌组织培养的技术要少。如果转基因根将用病原体攻击，则不需要用该技术产生无性病原体，因为不使用无菌组织培养基。因此，不存在其上可能生长污染物的琼脂培养基，导致模糊的结果或患病的根。在这里，我们描述使用发根土壤杆菌（Agrobacterium rhizogenes）在7日龄大豆幼苗上生产转基因大豆根。这些复合植物可以在温室中生长用于进一步实验，例如确定基因表达对线虫发育的影响。
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