To generate a reliable preclinical model system exhibiting the molecular 
features of salivary adenoid cystic carcinoma (ACC) whose biology is still 
unclear due to the paucity of stable cell cultures. To develop new in vitro and 
in vivo models of ACC, the techniques of organoid culture and patient-derived 
tumor xenograft (PDX), which have attracted attention in other malignancies in 
recent years, were applied. Tumor specimens from surgically resected salivary 
ACC were proceeded for the preparation of PDX and organoid culture. The 
orthotopic transplantation of patient-derived or PDX-derived organoids was 
demonstrated into submandibular glands of NSG mice and those histology was 
evaluated. PDX-derived organoid cells were evaluated for the presence of 
MYB-mediated fusion genes and proceeded for in vitro drug sensitivity assay. 
Human ACC-derived organoids were successfully generated in three-dimensional 
culture and confirmed the ability of these cells to form tumors by orthotopic 
injection. Short-term organoid cell cultures from two individual ACC PDX tumors 
were also established that maintain the characteristic MYBL1 translocation and 
histological features of the original parent and PDX tumors. Finally, the 
establishment of drug sensitivity tests on these short-term cultured cells was 
confirmed using three different agents. This is the first to report an approach 
for the generation of human ACC-derived organoids as in vitro and in vivo cancer 
models, providing insights into understanding of the ACC biology and creating 
personalized therapy design for patients with ACC.