This study aimed to promote gland formation in cells derived from endometrial cancer, and assess the relevance of sulfolipids by performing culture with type I collagen. Tumors were developed in nude mice using cultured cell lines, gland formation was induced by culture with type I collagen and the composition of tumor cell sulfolipids was analyzed. Results showed that after culturing the cells on type I collagen gel, the gel was floated. Another layer of gel was placed on top so that the cells were sandwiched between two layers. Using this method, it was possible to induce gland formation in cells that formed only poorly differentiated tumors in nude mice. Mucous staining and electron microscopy demonstrated polarity of the glands. The cell lines that showed gland formation expressed sulfolipids, but not cholesterol sulfate. In conclusion, type I collagen and sulfolipids are involved in the process of gland formation in endometrioid adenocarcinoma.