Fat grafting for breast cancer (BrCa) reconstruction and breast augmentation has become increasingly more popular. A major area of debate and controversy is the effect of adipose-derived stem cells (ASCs) on remnant or undetected BrCa cells. We investigate the in vitro response of BrCa to ASCs in a coculture model with regards to cell migration.The study was approved by the institutional review board. BrCa and adipose tissue specimens either from subcutaneous breast tissue or abdominal lipoaspirate were obtained from the same patient. BrCa cells and ASCs were harvested with either explant culture and/or enzymatic digestion. Tissues were grown in cell culture flasks until adequate cell libraries were established. Adipose-derived stem cells from adipose specimens were characterized with flow cytometry. Immunofluorescence (IF) staining of the initial cell population harvested from the BrCa specimens confirmed the presence of CD24, an epithelial marker of BrCa. A homogenous CD 24+/CD 90- BrCa cell population was obtained with flowcytometric cell sorting. The in vitro migration of BrCa cells was examined in coculture with and without ASCs.Adipose-derived stem cells harvested from the adipose specimens were positive for mesenchymal stem cell markers CD 105, CD 90, CD 73, and CD 44 and negative for lymphocyte cell marker CD 34 and leukocyte marker CD 45. The percentage of the CD 24+/CD 90- BrCa cells in the initial cell population harvested from BrCa specimens was 0.61%. The BrCa cells morphologically had large nuclei and small cytoplasm in clusters under the light microscope, suggesting a cancer cell phenotype. CD 24 expression on the surface of BrCa cells was confirmed with IF staining. The number of BrCa cells migrated in ASCs coculture was approximately 10 times higher than the number of BrCa cells migrated in BrCa cell only cultures.Adipose-derived stem cells significantly increase the migration capacity of BrCa cells in vitro in cocultures. This should be taken into consideration when performing fat grafting to the breast especially in patients with a history of BrCa or strong family history of BrCa.