The development of 3D cell culture techniques has begun to offer promising results for the field of cancer biology research in recent years, however, it is necessary to continue experimenting with protocols for tumor cells and to analyze their advantages or shortcomings with new Cell lines and / or material extracted from biopsies of different types of tumors for which culturing techniques of any kind have not yet been standardized. Therefore, this work aimed to establish a three-dimensional tumor culture model using the multicellular tumor spheroids (MTS) technique from tumor cell lines and biopsies of gastric and endometrial cancer tumors, establishing protocols For their sowing and analysis. The spheroids were cultured from the SiHa cell line, starting from monolayer cultures that were subsequently trypsinized to establish an ideal concentration that was seeded in wells coated with 1% agarose. With the spheroids already formed, tests are being carried out to make cuts that allow to observe the internal structure of the same, which are still in the analysis phase. At the end of the working time, it was possible to establish the protocol for the sowing of a three-dimensional cell culture using the multicellular tumor spheroid technique (MTS) from the SiHa cell line, as well as to seed these spheroids from the cultures Established primary tumor cells of gastric and endometrial cancer. This protocol opens the door to new research on gene silencing and virus penetration in more complex tumor systems.