Umbilical cord blood (UCB) transplantation is being used as an alternative source of hematopoietic stem cells for bone marrow reconstitution. Separation, processing and cry preservation of UCB samples in large numbers for storage in cord blood banks ideally needs to be partially automated to allow large numbers of samples to be processed efficiently. Aim of the present study was to carried out for standardization of collection, processing, cry preservation and quantification of CD34+ haematopoietic stem cells in umbilical cord blood. A total of 420 umbilical cord blood units were collected. Samples were analyzed for CD34+ cells concentration and viability of mononuclear cells were done on all the samples. The mean of the CD34+ cells concentration was 1.99± 0.82 % (range 0.4-4.5) on baseline period. The mean of the CD34+ cells concentration was 1.14± 0.85 % (range 0.2-3.8) on six month. The mean of the viability of mononuclear cells was 81.80 ±3.73℅ (range 76-91) on baseline period. The viability of mononuclear cells count at six month was 64.24± 2.70℅ (range 61-70). The viability loss of mononuclear cells from baseline to six month was 20.9℅. The mean of the total nucleated cells count was 11.36 ± 4.34 cells/ µl, (0.67- 16.68). In conclusion, the method demonstrates that UCB units can be routinely processed in a closed system that also achieves a significant reduction in storage needs and related costs, while maintaining quantity and quality of the haematopoitic stem cells.