An Optimal Condition for Red Cell Depletion of Cord Blood with Pentastarch

Lee, Young Ho; Park, Hyun Woo; Han, Hun; Han, Jin Yeong; Kim, Jae Seok; Kim, Hyo Jin

Abstract

PURPOSE: We evaluated the optimal temperature, from comparing at room temperature and at 4degrees C, on separation of cord blood with 10% pentastarch to increase the yield of progenitor cells. The yield of progenitor cells could be decreased in elapsed cord blood following collection, probably due to microclot or clumping as we observed in previous study. Thus, we also assessed the effect of urokinase on the recovery rate of progenitor cells in elapsed cord blood.
METHODS
We collected 11 samples of cord blood by syringes anticoagulated with ACD from the umbilical cord vein during or following vaginal delivery. The cord blood was separately stored at room temperature with and without urokinase (5,000 IU/mL), and processed with 10% pentastarch to sediment RBCs at room temperature and at 4degrees C, respectively. Total nucleated cell (TNC) and CD34+ cell counts of unseparated and separated cord blood were performed on 0,24 and 48 hours after being held at room temperature.
RESULTS
1) The recovery rate of TNC in the cord blood elapsed for 24 hours was 47.22+/-40.19% by processing at room temperature and 17.30+/-14.67% at 4degrees C (P=0.06). The recovery rate of CD34+ cell was 94.48+/-84.34% at room temperature and 27.30+/-33.44% at 4degrees C (P=0.05). There were no statistical significances in the cord blood elapsed for 48 hours. 2) There were no statistical differences of TNC and CD34+ cell count in the unseparated and separated cord blood regardless adding urokinase.
CONCLUSION
We suggest that it could minimize the loss of progenitor cells during processing to sediment red cells of cord blood with 10% pentastarch at room temperature rather than at 4degrees C. Further studies regarding the optimal dosage of urokinase to prevent clumping or microclot in elapsed cord blood and to increase the yield of progenitor cells would be required.