Mishra Dhruva K, Kim Min P
Department of Surgery, Houston Methodist Hospital Research Institute.
Department of Surgery, Houston Methodist Hospital Research Institute; Division of Thoracic Surgery, Department of Surgery, Weill Cornell Medical College, Houston Methodist Hospital;
J Vis Exp. 2018 Aug 19(138):58145. doi: 10.3791/58145.
It is difficult to isolate tumor cells at different points of tumor progression. We created an ex vivo lung model that can show the interaction of tumor cells with a natural matrix and continual flow of nutrients, as well as a model that shows the interaction of tumor cells with normal cellular components and a natural matrix. The acellular ex vivo lung model is created by isolating a rat heart-lung block and removing all the cells using the decellularization process. The right main bronchus is tied off and tumor cells are placed in the trachea by a syringe. The cells move and populate the left lung. The lung is then placed in a bioreactor where the pulmonary artery receives a continual flow of media in a closed circuit. The tumor grown on the left lung is the primary tumor. The tumor cells that are isolated in the circulating media are circulating tumor cells and the tumor cells in the right lung are metastatic lesions. The cellular ex vivo lung model is created by skipping the decellularization process. Each model can be used to answer different research questions.
在肿瘤进展的不同阶段分离肿瘤细胞是困难的。我们创建了一个离体肺模型,它可以展示肿瘤细胞与天然基质的相互作用以及营养物质的持续流动,还创建了一个展示肿瘤细胞与正常细胞成分和天然基质相互作用的模型。无细胞离体肺模型是通过分离大鼠心肺块并使用脱细胞过程去除所有细胞而创建的。右主支气管被结扎,肿瘤细胞通过注射器注入气管。细胞移动并在左肺中聚集。然后将肺置于生物反应器中,肺动脉在封闭回路中接受持续的培养基流动。在左肺上生长的肿瘤是原发性肿瘤。在循环培养基中分离出的肿瘤细胞是循环肿瘤细胞,右肺中的肿瘤细胞是转移病灶。细胞离体肺模型是通过跳过脱细胞过程而创建的。每个模型都可用于回答不同的研究问题。