We intend to grow human uterine tissue (myometrium) in three dimensions. We will use a novel scaffold-forming technique, and then seed cells into the scaffold. These structures will be made large enough to mimic normal human myometrium. This project will also demonstrate that this engineered tissue will function similarly to the normal tissue. We will show that the engineered tissue strips are capable of contracting, and the engineered uterus is capable of generating intracavitary pressure. We will develop tissue engineering as a new model for the study of contractions of the pregnant human uterus. This model will address major health questions of abnormal uterine function (such as preterm labor, post partum hemorrhage, and post-dates pregnancy). The novelty of the model is that both cellular gene expression and 3-D physical structure are capable of being controlled. We will start with human uterine tissue obtained from women at the time of Cesarean Delivery. We will amplify the number of cells by growing them using standard culture techniques. We will create scaffolds from chitosan, a readily available, naturally occurring, safe, bio-resorbable compound. These scaffolds can be made in virtually any shape and with a wide variety of porosities to support cell growth. We will determine the rates that cells grow into and around the scaffolds. We will compare the biochemical markers found in normal uterine tissue with those in the engineered tissue. We will compare the physiology of engineered tissue strips with strips recently removed from women. We will demonstrate, for the first time, generation of intracavitary pressure from a tissue-engineered uterus. In addition to creating a new paradigm for study of tissue-level and organ-level uterine function, these techniques are capable of being up-scaled to allow tissue patches to be created from a small smooth muscle sample from a patient. This project is the first step on the path to creating an entire replacement uterus for women born without a functional uterus, or who have had a hysterectomy. PUBLIC HEALTH RELEVANCE: Tissue-engineered myometrium will be made from human uterine cells and chitosan, a naturally occurring, resorbable scaffold material. The tissue-engineered myometrium will be a novel model for the study of myometrial contractility. This project will be the first step toward construction of a functional replacement uterus for women who were born with a uterine anomaly or have had a hysterectomy.