Department of Zoology, Genetics and Physical Anthropology, Universidade de Santiago de Compostela, Campus de Lugo, 27002 Lugo, Spain.
Genomic Medicine Group, Center for Research in Molecular Medicine and Chronic Diseases (CiMUS), Universidade de Santiago de Compostela, 15782 Santiago de Compostela, Spain.
Genes (Basel). 2021 Jan 19;12(1):113. doi: 10.3390/genes12010113.
Published studies show that most of the human cancer xenograft studies in zebrafish embryos have used incubation temperatures in the range of 32-34 °C for 3-6 days post-injection, trying to find a compromise temperature between the zebrafish embryos (28 °C) and the human injected cells (37 °C). While this experimental setup is widely used, a question remains: is possible to overcome the drawbacks caused by a suboptimal temperature for the injected cells? To clarify the effect of temperature and injected cells on the host, in this study, we analyzed the development and health of the last in response to different temperatures in the presence or absence of injected human cancer cells. Comparing different incubation temperatures (28, 34 and 36 °C), we determined morphological abnormalities and developmental effects in injected and non-injected embryos at different time points. Besides this, the expression of selected genes was determined by qPCR to determine temperature affected metabolic processes in the embryos. The results indicate that an incubation temperature of 36 °C during a period of 48 h is suitable for xenotransplantation without morphological or metabolic changes that could be affecting the host or the injected cells, allowing them to proliferate near their optimal temperature.
已发表的研究表明，大多数在斑马鱼胚胎中进行的人类癌症异种移植研究都使用了注射后 3-6 天、温度范围在 32-34°C 的孵化温度，试图在斑马鱼胚胎（28°C）和注射的人类细胞（37°C）之间找到一个折衷温度。虽然这种实验设置被广泛使用，但仍存在一个问题：是否有可能克服因注射细胞的温度不理想而导致的缺点？为了阐明温度和注射细胞对宿主的影响，在这项研究中，我们分析了最后一个的发育和健康状况，以了解在存在或不存在注入的人类癌细胞的情况下，温度的变化对宿主的影响。通过比较不同的孵化温度（28、34 和 36°C），我们在不同时间点确定了注射和未注射胚胎的形态异常和发育影响。除此之外，还通过 qPCR 确定了选定基因的表达，以确定温度是否影响胚胎中的代谢过程。结果表明，在 48 小时的时间内，36°C 的孵育温度适合于异种移植，而不会对宿主或注射细胞产生形态或代谢变化，从而使它们能够在接近最佳温度的条件下增殖。
