Cancer is the second leading cause of death worldwide, and triple-negative breast cancer (TNBC) patients show the poorest prognosis and survival and the highest metastasis prevalence among all breast cancer subtypes. Matcha has recently been associated with multiple health benefits, and in vitro studies showed the potential effect of matcha in inhibiting cancer development and metastasis. We aimed to determine the safe, non-toxic dose of matcha suitable for zebrafish and to investigate the anticancer effect of matcha on the metastasis and growth of human TBNC cells using a zebrafish xenograft model. Wild-type AB zebrafish were used to conduct multiple general toxicity assessments, including developmental, neuromuscular, and cardiovascular toxicities. The safe, non-toxic concentration of matcha was determined to be 50 µg/mL and 100 µg/mL. Afterward, the zebrafish xenograft model was successfully established for MDA-MB-468 and MDA-MB-231 TNBC cells. The tumor size and metastasis of the injected cancer cells were traced through CM-Dil red fluorescent dye. Upon exposure to matcha at the safe doses, MDA-MB-231 and MDA-MB-468 showed a trend toward reduction in tumor size in a dose-dependent manner, indicated by quantified fluorescence. Matcha also visibly suppressed metastasis of cancer cells in the zebrafish body. Our results point to a potential dose-dependent anticancer effect of matcha on TNBC cells; however, more extended observation periods after xenotransplantation are required to confirm the long-term anticancer effect of matcha on tumor growth and metastasis.