Christina Basford1, Nicolas Forraz2, Saba Habibollah1, Kendal Hanger1, Colin McGuckin2. 1. Newcastle Centre for Cord Blood, Institute of Human Genetics, Newcastle University, Newcastle upon Tyne, NE1 3BZ, United Kingdom. 2. CTI-LYON, Cell Therapy Research Institute, Parc Technologique de Lyon-St Priest, Cèdre 1, 97 Allée Alexandre Borodine, 69800 SAINT PRIEST LYON, France.
Abstract
BACKGROUND AND OBJECTIVES: Well over 1 million Umbilical Cord Blood units (UCB) have been stored globally in the last 10 years. Already, over 20,000 transplants been performed using UCB for haematopoietic reconstitution alone, now this potential is joined by regenerative medicine. However, more needs to be known about processing of this stem cell source for it to reach full potential. METHODS AND RESULTS: IN THIS STUDY WE EVALUATED FIVE SEPARATION METHODS: plasma depletion, density gradient, Hetastarch, a novel method known as PrepaCyte-CB and an automated centrifugal machine. Sepax gives the highest recovery of nucleated cells, an average of 78.8% (SD±21.36). When looking at CD34+ haematopoietic stem cells PrepaCyte-CB provided the greatest recovery at 74.47% (SD±8.89). For volume reduction density gradient was the most effective leaving 0.03×10(6) RBC/ml, 8 times more efficient than its nearest competitor PrepaCyte-CB (p<0.05). Finally PrepaCyte-CB processing left samples with the highest clonogenic potential after processing and more significantly after cryopreservation: 9.23 CFU/10(8) cells (SD±2.33), 1.5 fold more effective than its nearest rival Sepax (p<0.05). CONCLUSIONS: PrepaCyte-CB was the most flexible method; the only processing type unaffected by volume. Results indicate that processing choice is important depending on your final intended use.
BACKGROUND AND OBJECTIVES: Well over 1 million Umbilical Cord Blood units (UCB) have been stored globally in the last 10 years. Already, over 20,000 transplants been performed using UCB for haematopoietic reconstitution alone, now this potential is joined by regenerative medicine. However, more needs to be known about processing of this stem cell source for it to reach full potential. METHODS AND RESULTS: IN THIS STUDY WE EVALUATED FIVE SEPARATION METHODS: plasma depletion, density gradient, Hetastarch, a novel method known as PrepaCyte-CB and an automated centrifugal machine. Sepax gives the highest recovery of nucleated cells, an average of 78.8% (SD±21.36). When looking at CD34+ haematopoietic stem cells PrepaCyte-CB provided the greatest recovery at 74.47% (SD±8.89). For volume reduction density gradient was the most effective leaving 0.03×10(6) RBC/ml, 8 times more efficient than its nearest competitor PrepaCyte-CB (p<0.05). Finally PrepaCyte-CB processing left samples with the highest clonogenic potential after processing and more significantly after cryopreservation: 9.23 CFU/10(8) cells (SD±2.33), 1.5 fold more effective than its nearest rival Sepax (p<0.05). CONCLUSIONS:PrepaCyte-CB was the most flexible method; the only processing type unaffected by volume. Results indicate that processing choice is important depending on your final intended use.
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