Cell recovery sufficient for adult transplantation by additional cord blood collection from placenta

The amount of newborn blood that can be collected from a single cord donor is limited, but a significant amount remains in the placenta. We used a simplified perfusion method to collect this additional blood. Umbilical cord blood from 15 newborns was collected before placental delivery by umbilical vein puncture. After delivery, the placenta was placed on sterile gauze and 63 mL of citrate-phosphate-dextrose-adenine anticoagulant were injected into the umbilical vein that was then clamped near the placenta. The placenta was gently massaged, hung over a sterile vessel, and the umbilical cord cut sterilely near the embryonic surface. Additional blood was collected into the sterile vessel by pressuring a gauze bag around the placenta. We assessed the contribution of this second fraction to the total volume, total nucleated cell (NC), CD34+, hematopoietic progenitors cell, and colony forming unit count and bacterial contamination risk. The total collected volume was 127.3 mL (range 92-170) and the NC content was 1.6+/-0.73x10(9). The mean second fraction contribution from 15 units to the total nucleated and mononuclear cell content was 54+/-9.87% and 54+/-9.52%, respectively. The added percentage of CD34+ and hematopoietic progenitor cells was 54.3+/-10.35% and 46.7+/-11.5%, respectively, while the additional percentages of colony forming-granulocyte macrophage and colony forming-erythroid in the second fraction were 43.2+/-5.5% and 39.8+/-4.3%, respectively, indicating that the cells collected after placental perfusion (second fraction) had similar HPC content and in vitro hematopoietic potential. The method did not increase the risk of bacterial contamination.