Zen-Bio, Inc focuses on the in vitro isolation and culture of human adipose tissues and has an international reputation in this field. There is an unmet medial need for new technologies to create implant adipose tissues. Lipodystrophy patients lose subcutaneous fat, leading to life threatening diabetes. Creating new adipose tissue would simplify their medical management. Breast cancer patient routinely undergo a transverse rectus abdominal musculocutaneous procedure after mastectomy to achieve breast reconstruction. Many of these same patients could undergo a less costly and invasive "lumpectomy" but elect not to since the outcome is aesthetically unacceptable. Bio-engineered autologous fat to repair lumpectomies would encourage more of these procedures. In the proposed experiments, Zen-Bio Inc. will obtain" proof of principle" data demonstrating that human adipose tissue derived stromal cells can be used for soft tissue reconstruction and transplanting. Specific Aim I will further optimize cryopreservation of human adipose-derived stromal cells and maintain their differentiation potential in vitro. Specific Aim 2 will determine the ability of human adipose-derived stromal cells to form a fat pad when implanted subcutaneously in immunodeficient mice. These studies will demonstrate the commercial feasibility of our core hypothesis, that autologous adipose derived stromal cells can be used for soft tissue reconstruction