In this study, three-dimensional bio-tissue scaffolds were prepared in vitro, which play an important role in tissue engineering. The aim of this research was to study the use of sheep omentum in the manufacture of soft tissue engineering scaffolds used in the laboratory, as this biosynthesis is characterized by its availability and cheap price. The biological metabolism of scaffolds also plays an important role in the process of new tissue formation. A porous biomaterial scaffold was designed using connective tissue from sheep omentum, which was successfully prepared by a freeze-drying method that included a decellularization process and the process of removing cells was confirmed using DAPI staining. The following properties of the scaffold were then measured, such as the biological properties of the resulting tissue, as well as Glycosaminoglycans (GAG) and metabolic activity (MTT) tests which proved that this tissue formed was not toxic to the cells seeded on it , structural properties, mechanical tests, and microscopy . Scanning electron imaging (SEM).