Human umbilical cord blood (UCB) is a valuable alternative source of ethically acceptable, clinically competent stem cells that is most likely closest to embryonic stem cells. Development of reliable methods for the expansion of cord blood stem cells is critical to ensure their clinical application. In the present article, advances in cord blood stem cell isolation, culture expansion methods through co-culture with human mesenchymal stem cells, culture optimization techniques with defined media and cord blood stem cell banking aspects have been reviewed. Refined methods of isolation as well as defined culture conditions of expansion that favor retention of stem cell phenotype and proper cryogenic storage can significantly increase the use of cord blood stem cells in human cell therapy applications.