Casey Mattie J, Modzelewska Katarzyna, Anderson Daniela, Goodman James, Boer Elena F, Jimenez Laura, Grossman Douglas, Stewart Rodney A
Department of Oncological Sciences and Huntsman Cancer Institute, University of Utah School of Medicine.
Department of Oncological Sciences and Huntsman Cancer Institute, University of Utah School of Medicine; Department of Dermatology, University of Utah Health Sciences Center, Salt Lake City.
J Vis Exp. 2017 May 17(123):55712. doi: 10.3791/55712.
Tumor cell transplantation is an important technique to define the mechanisms controlling cancer cell growth, migration, and host response, as well as to assess potential patient response to therapy. Current methods largely depend on using syngeneic or immune-compromised animals to avoid rejection of the tumor graft. Such methods require the use of specific genetic strains that often prevent the analysis of immune-tumor cell interactions and/or are limited to specific genetic backgrounds. An alternative method in zebrafish takes advantage of an incompletely developed immune system in the embryonic brain before 3 days, where tumor cells are transplanted for use in short-term assays (i.e., 3 to 10 days). However, these methods cause host lethality, which prevents the long-term study of tumor cell behavior and drug response. This protocol describes a simple and efficient method for the long-term orthotopic transplantation of zebrafish brain tumor tissue into the fourth ventricle of a 2-day-old immune-competent zebrafish. This method allows: 1) long-term study of tumor cell behaviors, such as invasion and dissemination; 2) durable tumor response to drugs; and 3) re-transplantation of tumors for the study of tumor evolution and/or the impact of different host genetic backgrounds. In summary, this technique allows cancer researchers to assess engraftment, invasion, and growth at distant sites, as well as to perform chemical screens and cell competition assays over many months. This protocol can be extended to studies of other tumor types and can be used to elucidate mechanisms of chemoresistance and metastasis.
肿瘤细胞移植是一种重要的技术，可用于确定控制癌细胞生长、迁移和宿主反应的机制，以及评估患者对治疗的潜在反应。目前的方法很大程度上依赖于使用同基因或免疫缺陷动物，以避免肿瘤移植被排斥。此类方法需要使用特定的基因品系，这往往会妨碍对免疫-肿瘤细胞相互作用的分析，和/或仅限于特定的基因背景。斑马鱼中的另一种方法利用了3天前胚胎大脑中发育不完全的免疫系统，在此处移植肿瘤细胞用于短期分析（即3至10天）。然而，这些方法会导致宿主死亡，从而妨碍对肿瘤细胞行为和药物反应的长期研究。本方案描述了一种简单有效的方法，用于将斑马鱼脑肿瘤组织长期原位移植到2日龄具有免疫能力的斑马鱼的第四脑室。该方法允许：1）长期研究肿瘤细胞行为，如侵袭和扩散；2）肿瘤对药物的持久反应；3）肿瘤的再移植，用于研究肿瘤进化和/或不同宿主基因背景的影响。总之，该技术使癌症研究人员能够评估远处部位的植入、侵袭和生长情况，并在数月内进行化学筛选和细胞竞争分析。本方案可扩展到其他肿瘤类型的研究，并可用于阐明化疗耐药性和转移的机制。
