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Human umbilical cord blood (HUCB) is now considered as a valuable source for stem cell-based therapies. Previous studies showed that intravascular injection of the HUCB significantly improves neurological functional recovery in a model of intracerebral hemorrhage (ICH). To extend these findings, we examined the behavioral recovery and injured volume in the presence of increasing doses of human umbilical cord blood derived mononuclear cells (HUC-MCs) after intracerebral hemorrhage in rats. The experimental ICH was induced by intrastriatal administration of bacterial collagenase IV in adult rats. One day after the surgery, the rats were randomly divided into 4 groups to receive intravenously either BrdU positive human UC-MCs (4 × 10(6), 8 × 10(6) and 16 × 10(6) cells in 1 ml saline, n=10, respectively) as treated groups or the same amount of saline as lesion group (n=10). There was also one group (control n=10) that received only the vehicle solution of collagenase. The animals were evaluated for 14 days with modified limb placing and corner turn tests. The transplanted human UC-MCs were also detected by immunohistochemistry with labeling of BrdU. Two weeks after infusion, there was a significant recovery in the behavioral performance when 4 × 10(6) or more UC-MCs were delivered (P<0.05-0.001). Injured volume measurements disclosed an inverse relationship between UC-MCs dose and damage reaching significance at the higher UC-MCs doses. Moreover, human UC-MCs were localized by immunohistochemistry only in the injured area. Intravenously transplanted UC-MCs can accelerate the neurological function recovery of ICH rat and diminish the striatum lesion size by demonstrating a dose relationship between them., (Copyright © 2012 Elsevier GmbH. All rights reserved.)

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.