Coats Jacqueline S, Baez Ineavely, Stoian Cornelia, Milford Terry-Ann M, Zhang Xiaobing, Francis Olivia L, Su Ruijun, Payne Kimberly J
Department of Pathology and Human Anatomy, Loma Linda University.
Department of Basic Sciences, Loma Linda University.
J Vis Exp. 2017 May 10(123):55384. doi: 10.3791/55384.
Patient-derived xenograft (PDX) mice are produced by transplanting human cells into immune deficient mice. These models are an important tool for studying the mechanisms of normal and malignant hematopoiesis and are the gold standard for identifying effective chemotherapies for many malignancies. PDX models are possible because many of the mouse cytokines also act on human cells. However, this is not the case for all cytokines, including many that are critical for studying normal and malignant hematopoiesis in human cells. Techniques that engineer mice to produce human cytokines (transgenic and knock-in models) require significant expense before the usefulness of the model has been demonstrated. Other techniques are labor intensive (injection of recombinant cytokine or lentivirus) and in some cases require high levels of technical expertise (hydrodynamic injection of DNA). This report describes a simple method for generating PDX mice that have exogenous human cytokine (TSLP, thymic stromal lymphopoietin) via weekly intraperitoneal injection of stroma that have been transduced to overexpress this cytokine. Use of this method provides an in vivo source of continuous cytokine production that achieves physiological levels of circulating human cytokine in the mouse. Plasma levels of human cytokine can be varied based on the number of stromal cells injected, and cytokine production can be initiated at any point in the experiment. This method also includes cytokine-negative control mice that are similarly produced, but through intraperitoneal injection of stroma transduced with a control vector. We have previously demonstrated that leukemia cells harvested from TSLP-expressing PDX, as compared to control PDX, exhibit a gene expression pattern more like the original patient sample. Together the cytokine-producing and cytokine-negative PDX mice produced by this method provide a model system that we have used successfully to study the role of TSLP in normal and malignant hematopoiesis.
患者来源的异种移植(PDX)小鼠是通过将人类细胞移植到免疫缺陷小鼠体内产生的。这些模型是研究正常和恶性造血机制的重要工具,也是鉴定许多恶性肿瘤有效化疗方法的金标准。PDX模型之所以可行,是因为许多小鼠细胞因子也作用于人类细胞。然而,并非所有细胞因子都是如此,包括许多对研究人类细胞正常和恶性造血至关重要的细胞因子。使小鼠产生人类细胞因子的技术(转基因和敲入模型)在证明模型的实用性之前需要大量费用。其他技术劳动强度大(注射重组细胞因子或慢病毒),在某些情况下需要高水平的技术专长(DNA的流体动力学注射)。本报告描述了一种生成PDX小鼠的简单方法,即通过每周腹腔注射已被转导以过表达这种细胞因子的基质细胞,使PDX小鼠具有外源性人类细胞因子(TSLP,胸腺基质淋巴细胞生成素)。使用这种方法可提供体内持续产生细胞因子的来源,在小鼠体内实现循环人类细胞因子的生理水平。人类细胞因子的血浆水平可根据注射的基质细胞数量而变化,并且细胞因子的产生可在实验的任何时间点开始。该方法还包括类似产生的细胞因子阴性对照小鼠,但通过腹腔注射用对照载体转导的基质细胞。我们之前已经证明,与对照PDX相比,从表达TSLP的PDX收获的白血病细胞表现出更类似于原始患者样本的基因表达模式。通过这种方法产生的产生细胞因子和细胞因子阴性的PDX小鼠共同提供了一个模型系统,我们已成功使用该系统研究TSLP在正常和恶性造血中的作用。