Cell culture technology plays an essential role in modern biomedical research, especially in cancer research. The Cell Culture Facility (CCF) serves as a customized pay-for-service Facility that provides expertise, technical support, reagents, equipment, liquid nitrogen storage, and/or training to increase the effectiveness and efficiency of using mammalian cell culture in FCCC laboratories. This Facility provides technical assistance and consultation services in tissue culture techniques, centralized liquid nitrogen banking, mycoplasma screening services, the preparation of custom and standard culture media and supplements, performance testing of fetal bovine serum, cell propagation, and technical support and supplies for mouse embryonic stem (ES) cell culture. Major efforts by the CCF include carrying out technically-challenging techniques, such as the establishment of primary cell cultures from patient or animal tissue samples and the generation of gene "knockout" or "knockin" ES cells. In 2003, the Facility served 52 investigators with peer-reviewed funding and a total of 56 of the 65 research laboratories at FCCC from all three Divisions and 11 Research Programs. Peer-reviewed, funded research accounted for more than 92.5% of CCF usage. The Facility is highly-regarded among faculty users, and received an "Outstanding to Excellent" rating at the last CCSG review. In the past five years, the demand for CCF services has increased substantially, with 225% more primary cell lines established and 54-117% increases in the distribution of media, sterile reagents, and serum. Importantly, the Facility supplies the FCCC community with valuable on-site expertise for a host of established cell culture protocols, consultation in the design and evaluation of experiments, training in cell culture methodologies, and adaptable assistance delivered in a cost-effective manner. The Facility continues to provide innovative services, including the recent establishment of "nucleofection" services to efficiently transfect cell lines and the production of extracellular matrices for "three-dimensional" cell culture. Thus, the CCF serves FCCC investigators with a ready source of expertise, manpower, and a convenient and cost-effective resource for culture supplies and specialized services.