TUMOR-ALIGNED COCULTURE SYSTEM

许多抗癌药物在人体试验中失败，尽管在体外研究和动物模型中显示出疗效。已经清楚的是，涉及2D单一培养的体外测定不能反映肿瘤组织的复杂细胞和基质微环境，这可以解释2D模型预测临床疗效的失败。本研究的目的是使用乳腺癌、基质细胞和内皮细胞开发代表人类肿瘤组织结构的体外肿瘤对齐3D共培养模型，并使用已知有效的抗癌药物测试该模型。第一个目标是优化细胞外基质的组成和细胞模型，以引起生理肿瘤形态，增殖，侵袭，基质募集和内皮发芽。该合同的第二个目标是使用RT-PCR，Western Blot和免疫细胞化学来表征该模型与传统的2D和3D培养模型相比的基因表达变化。本合同的第三个目标是通过评估氟尿嘧啶和紫杉醇的生理特性来验证模型的有效性。这种模式的成功实施将大大节省药物开发的成本和时间，最终造福患者和社会。

Many anticancer drugs fail in human trials despite showing efficacy in in vitro studies and animal models. It has become clear that the in vitro assays involving 2D monoculture do not reflect the complex cellular and matrix microenvironment of the tumor tissue and this may explain the failure of 2D models to predict clinical efficacy. The goal of this study is to develop an in vitro tumor-aligned 3D coculture model representative of the human tumor tissue architecture using breast cancer, stromal, and endothelial cells and test this model with known effective anticancer drugs. The first objective it to optimize the composition of the extracellular matrix and the cell model to elicit the physiogical tumor morphology, proliferation, invasion, stromal recruitment, and endothelial sprouting . The second objective of this contract is to characterize changes in gene expression for this model compared to traditional 2D and 3D culture models using RT-PCR, Western Blot, and Immunocytochemistry. The third objective of this contract is to validate the model as effective by evaluating these physiological properties in response to Fluorouracil and Paclitaxel. Successful implementation of this model would save significant cost and time for drug development, ultimately benefiting the patient and society.