The present invention relates to fed batch culture methods for high cell density growth of Listeria which produce cultures having an OD600 greater than about 2.2 or higher. In particular, the invention provides methods for high cell density growth of Listeria comprising growth in a pH controlled bioreactor and, optionally, the gradual addition of a carbon source, e.g., glucose, with or without one or more additional nutrients, e.g., vitamins, when growth in the initial culture is nearly complete or complete. In one embodiment, the methods of the invention are used to produce Listeria-based compositions, e.g., vaccines comprising Listeria that express a tumor-associated antigen, e.g., an EphA2 antigenic peptide, for eliciting an immune response against hyperproliferative cells.