J M Alonso1, D M Regan, C E Johnson, D A Oliver, R Fegan, L C Lasky, D A Wall. 1. Division of Pediatric Hematology/Oncology, Department of Pediatrics, Pediatric Research Institute, Cardinal Glennon Children's Hospital/Saint Louis University, St Louis, MO, USA.
Abstract
BACKGROUND: In UC blood banking, volume and RBC reduction of the collected UC blood allows more efficient long-term storage and decreases infusion-related hemolysis and DMSO toxicity. However, high cell yields are imperative. At the St Louis Cord Blood Bank, we have systematically addressed processing/freezing and have developed a simple processing/freezing procedure. METHOD: The methodology is a modification of the hetastarch sedimentation and volume reduction approach of Rubinstein at the New York Placental Blood Program. Cord blood is mixed with a 1:5 v/v ratio of hetastarch. The product is incubated for 45 min in an inverted position in a refrigerated centrifuge (4 degrees C), and then is spun for 5 min at 50 g. RBC concentrate is drained from the bottom. The volume drained is calculated to remove 80% of RBC. The UC blood unit is then resuspended and spun for 13 min at 420 g. Plasma is expressed from the top. RESULTS: A final product volume of 27 mL (range 16-58 mL) was obtained from an original 50-200 mL of UC blood collected. The average yield of total nucleated cells pre- and post-processing was 90% for the first 4055 UC blood units banked. Pre- and post-processing CFU and CD34 yields were tested in a cohort and were similarly conserved. With a processing time of 3 h for a single cord, this process is time efficient and lends itself well to processing several units at the same time. The technique has been exported to other laboratories with similar yields. DISCUSSION: This simple methodology results in reliable yields and is well suited to larger scale banking.
BACKGROUND: In UC blood banking, volume and RBC reduction of the collected UC blood allows more efficient long-term storage and decreases infusion-related hemolysis and DMSOtoxicity. However, high cell yields are imperative. At the St Louis Cord Blood Bank, we have systematically addressed processing/freezing and have developed a simple processing/freezing procedure. METHOD: The methodology is a modification of the hetastarch sedimentation and volume reduction approach of Rubinstein at the New York Placental Blood Program. Cord blood is mixed with a 1:5 v/v ratio of hetastarch. The product is incubated for 45 min in an inverted position in a refrigerated centrifuge (4 degrees C), and then is spun for 5 min at 50 g. RBC concentrate is drained from the bottom. The volume drained is calculated to remove 80% of RBC. The UC blood unit is then resuspended and spun for 13 min at 420 g. Plasma is expressed from the top. RESULTS: A final product volume of 27 mL (range 16-58 mL) was obtained from an original 50-200 mL of UC blood collected. The average yield of total nucleated cells pre- and post-processing was 90% for the first 4055 UC blood units banked. Pre- and post-processing CFU and CD34 yields were tested in a cohort and were similarly conserved. With a processing time of 3 h for a single cord, this process is time efficient and lends itself well to processing several units at the same time. The technique has been exported to other laboratories with similar yields. DISCUSSION: This simple methodology results in reliable yields and is well suited to larger scale banking.
Authors: Allison Hubel; Ralf Spindler; Julie M Curtsinger; Bruce Lindgren; Sara Wiederoder; David H McKenna Journal: Transfusion Date: 2014-12-19 Impact factor: 3.157