Multicellular tumor spheroids (MCTS) demonstrate a physiologically relevant microenvironment, similar growth kinetics, and almost identical cell-cell and cell-extracellular matrix interaction as those in vivo. MCTS can be cultured in a laboratory with various techniques; one of them is the liquid overlay technique that has not been performed in Asian oral cancer cell lines; ORL-48 and ORL-136. This technique is inexpensive and straightforward, with ease of maintenance and easy access to spheroids. The cancer cells are inhibited from adherence to the culture plates’ surface by the liquid overlay technique to enable 3-dimension (3D) spheroid formation. This study aimed to grow and characterize the 3D spheroids of ORL-48 and ORL-136 with the liquid overlay technique. 10,000 and 25,000 cells per well of ORL-48 and ORL-136 were grown for seven days in U-bottomed 96-well plates pre-coated with 1% agarose. The characterization of spheroids’ morphology, including shape, size, growth, surface characteristics, and core structure, was evaluated. The spheroids of two cell lines demonstrated different sizes but similar shape and growth characteristics. The spheroids of both cell lines showed a greater diameter with a cell density of 25,000 cells/well compared with a cell density of 10,000 cells/well. Besides, ORL-136 spheroids showed relatively smaller diameters for both cell densities as compared with ORL-48 spheroids. The spheroids became more compact on day 7 as compared with day 1. The hematoxylin and eosin-stained sections of the spheroids demonstrated densely packed sheets of pleomorphic squamous epithelial cells at their cores and 2-3 layers of squamous epithelial cells at their periphery. In conclusion, ORL-48 and ORL-136 spheroids were successfully developed via the liquid overlay technique. This model can be further studied to utilize for in vitro anticancer drug screening and could provide a suitable platform for future research on novel anticancer drugs’ cytotoxicity.