Processing, Cryopreservation and Culture of Hematopoietic Precursors for Umbilic Cord Blood Bank

[Objective] To investigate the conditions on processing and cryopreservation of umbilic cord blood (UCB) and expansion of hematopoietic precursors ex vivo for wide establishment of UCB bank. [Methods] We designed the procedure based on FEES density gradient centrifugation and compared two strategies for UCB collection and processing. [Results] The two strategies could get enriched nucleated cells. The average yield of total nucleated cells and the volume reducing in closed-bag system was higher than that in centrifuge tubes, even the latter was more convenient and less expensive. The potent proliferative ability of hematopoietic stem and progenitor cells was detected by the clone forming unit granulocyte-macrophage (CFU-GM) culture of post-processing and thawing nucleated cells. The effect of cryopreservation was also well proved by the recovery of thawing cells. [Conclusion] The procedure based on HES density gradient centrifugation and the cryopreservation method we designed provided the foundation of an effective and convenient procedure of collection, processing and preservation of UCB for UCB bank word-widely.