Cord blood (CB) provides an alternative source of stem cells for transplantation, although in a considerable number of cases CB transplantation is followed by long periods of aplasia. Ex vivo expansion has the capacity to generate large amounts of progenitors, and it has been proposed that expanded cells might be beneficial in overcoming these long periods of aplasia. We describe the biological characteristics of cord blood compared to other sources of stem cells (BM and PB), and report the effects of FLT3-L and MIP-1alpha when added to a combination of SCF, IL-3 and IL-6 in pre-clinical short-term, serum-free expansion cultures of CB-derived CD34+ cells. After 6 days, this culture system was able to generate considerable expansion rates in the committed compartment (between 8.16- and 17.26-fold for CFU-GM, and 21.58- and 36.53-fold for the BFU-E/CFU-Mix), and the CD34+ population (between 11.25- and 25.42-fold). Moreover, this culture system was also able to maintain the week 5 CAFC population, particularly when both FLT3-L and MIP-1alpha were present (91% of the input level). Thus, we have described a pre-clinical protocol for ex vivo expansion of CB CD34+ cells in a short-term, static, serum-free system, where a high generation of committed progenitor cells is achieved together with CAFC maintenance.