- Email: [email protected]
recovery, or recipient engraftment
Poster Abstracts
performed manually but is labor intensive, operator dependent and increases the risk of product contamination. Protocols for automated cell processing systems have been developed for washing CB with DA. We present the results of our analysis comparing cellular recoveries achieved after manual or automated wash. Methods: CBUs included in this analysis were distributed by the Carolinas Cord Blood Bank (CCBB) for use in UCBT in the Duke Pediatric and Adult Transplant Programs, were washed manually utilizing the Rubinstein method (PNAS 1995; washed during 1999–2004; N = 91) or using the automated Sepax system (Biosafe International; Protocol v308; washed during 2010–2016; N = 62). Both protocols use DA at a 1:8 dilution. Data collection included precryopreservation TNCC, total CD34+ and CFUs. All assays were performed by the Duke Stem Cell Laboratory with exception of pre-cryopreservation TNCC data (performed by CCBB). Recovery of TNCC, total CD34+ and CFUs after thawing and washing were compared based on wash method using the Wilcoxon rank sum test. Results: Similar TNCC and total CD34+ recoveries were observed after a manual or automated wash (Table I). Significantly higher CFU recoveries occurred in CBUs washed using the Sepax automated cell processing system (P = 0.009; Figure 1). Conclusion: In this study, we compared cellular recoveries of thawed CBUs washed either manually or using the Sepax automated cell processor. While the TNCC and total CD34+ recoveries were similar between the two methods, CBUs washed via an automated system had higher CFU recovery. Since CFUs are an important measure of CBU potency and predictor of engraftment, this finding favors use of an automated wash method over a manual one. Ultimately, utilizing an automated wash is a safe, efficient, and feasible way of processing CBU. 121 HPC, APHERESIS PRODUCTS WITH HIGH WHITE BLOOD CELL CONCENTRATIONS CAN BE SAFELY STORED OVERNIGHT PRIOR TO CRYOPRESERVATION WITHOUT IMPACTING CD34+ CELL VIABILITY/RECOVERY, OR RECIPIENT ENGRAFTMENT N.L. Prokopishyn1,2, S. Berrigan1, A. Hillman1, H. Stastna1, N. Favell1 1 Cellular Therapy Laboratory, Calgary Laboratory Services, Calgary, Alberta, Canada, 2Pathology and Laboratory Medicine, University of Calgary, Calgary, Alberta, Canada Autologous transplant is a crucial treatment step mqny patientw. Successful collection and cryopreservation of HPC, Apheresis products (HPC (A)) is essential to successful transplant outcome. A high quality cryopreserved HPC (A) with adequate recovery and viability of CD34+ cells is essential to engraftment. Some studies have indicated decreased CD34+ cell viability/recovery in products that are stored at 40C prior to cryopreservation when the White Blood Cell (WBC) concentration ([WBC]) is higher than 200 × 109/L. Processing facilities often either cryopreserve product immediately following collection (as a late shift) or dilute product to a final [WBC] of 200 × 109/L prior to storage. Historical
Figure 1.
S59
data in our centre showed no evidence high [WBC] in collected products impacted CD34+ cell recovery or functionality following overnight storage. Therefore, temporary storage of collected products at 4°C without dilution of products to the [WBC] of 200 × 109/L is permited. With the recent updates in collection procedures, we felt that it prudent to re-examine [WBC] influence on CD34+ cell recovery/viability post-cryopreservation and thaw. HPC (A) collections (n = 369) were analyzed to detemine if [WBC] of product post collection impacted cell recovery and viability. All products were cryopreserved the day after collection following overnight storage at 40C. Products were cryopreserved by controlled rate freezer protocol and stored in vapour phase liquid nitrogen freezers until thaw and transplant. Quality Control (QC) vials representative of product were thawed and enumerated for total CD34+ cells surviving and % CD34+ cell viability (7-AAD). Percent (%) CD34+ cell recovery, % viable CD34+ cells, and % WBC recovery were calculated for all products and compared to collected [WBC]. There was no observed difference in % CD34+ cell recovery or viability in the products with Low [WBC] (<200 × 109/L) and High [WBC] (>200 × 109/L). As well, patients receiving products from the “low” and “high” [WBC] groups demonstrated no difference in time to neutrophil or platelet engraftment indicating equivalent functionality of the products. In summary, these findings demonstrate that neither CD34+ cell recovery, viability, nor functionality are impacted by [WBC] higher than 200 × 109/L when HPC (A) products are stored overnight prior to cryopreservation.
122 WILL NOT BE PRESENTED
123 PROLONGED TRANSIT TIME DOES NOT IMPAIR HPC MANIPULATION AND CRYOPRESERVATION P.J. Shaw1,2,3, V. Antonenas1,2, K. Micklethwaite1,2, L.E. Clancy2, K. Yehson1, D. Tong1, D. Gottlieb1,2 1 The Westmead and Children’s Hospital BMT Laboratory, Westmead Hospital, Westmead, New South Wales, Australia, 2Sydney Cellular Therapies Laboratory, Westmead Institute for Medical Research, Westmead, New South Wales, Australia, 3BMT, The Children’s Hospital at Westmead, Sydney, New South Wales, Australia Introduction: In Australia, we commonly obtain cells which have been transported long distances, with significant time delays prior to transplantation, and we have recently reported this. (Patton et al BBMT in press). At The Children’s Hospital, Westmead, it is our standard practice to CD34 select unrelated HPC products ahead of transplant. For this reason, we studied the time taken for the HPC cells to reach our facility and analysed the recovery and purity of CD34 positive cells after selection.
performed manually but is labor intensive, operator dependent and increases the risk of product contamination. Protocols for automated cell processing systems have been developed for washing CB with DA. We present the results of our analysis comparing cellular recoveries achieved after manual or automated wash. Methods: CBUs included in this analysis were distributed by the Carolinas Cord Blood Bank (CCBB) for use in UCBT in the Duke Pediatric and Adult Transplant Programs, were washed manually utilizing the Rubinstein method (PNAS 1995; washed during 1999–2004; N = 91) or using the automated Sepax system (Biosafe International; Protocol v308; washed during 2010–2016; N = 62). Both protocols use DA at a 1:8 dilution. Data collection included precryopreservation TNCC, total CD34+ and CFUs. All assays were performed by the Duke Stem Cell Laboratory with exception of pre-cryopreservation TNCC data (performed by CCBB). Recovery of TNCC, total CD34+ and CFUs after thawing and washing were compared based on wash method using the Wilcoxon rank sum test. Results: Similar TNCC and total CD34+ recoveries were observed after a manual or automated wash (Table I). Significantly higher CFU recoveries occurred in CBUs washed using the Sepax automated cell processing system (P = 0.009; Figure 1). Conclusion: In this study, we compared cellular recoveries of thawed CBUs washed either manually or using the Sepax automated cell processor. While the TNCC and total CD34+ recoveries were similar between the two methods, CBUs washed via an automated system had higher CFU recovery. Since CFUs are an important measure of CBU potency and predictor of engraftment, this finding favors use of an automated wash method over a manual one. Ultimately, utilizing an automated wash is a safe, efficient, and feasible way of processing CBU. 121 HPC, APHERESIS PRODUCTS WITH HIGH WHITE BLOOD CELL CONCENTRATIONS CAN BE SAFELY STORED OVERNIGHT PRIOR TO CRYOPRESERVATION WITHOUT IMPACTING CD34+ CELL VIABILITY/RECOVERY, OR RECIPIENT ENGRAFTMENT N.L. Prokopishyn1,2, S. Berrigan1, A. Hillman1, H. Stastna1, N. Favell1 1 Cellular Therapy Laboratory, Calgary Laboratory Services, Calgary, Alberta, Canada, 2Pathology and Laboratory Medicine, University of Calgary, Calgary, Alberta, Canada Autologous transplant is a crucial treatment step mqny patientw. Successful collection and cryopreservation of HPC, Apheresis products (HPC (A)) is essential to successful transplant outcome. A high quality cryopreserved HPC (A) with adequate recovery and viability of CD34+ cells is essential to engraftment. Some studies have indicated decreased CD34+ cell viability/recovery in products that are stored at 40C prior to cryopreservation when the White Blood Cell (WBC) concentration ([WBC]) is higher than 200 × 109/L. Processing facilities often either cryopreserve product immediately following collection (as a late shift) or dilute product to a final [WBC] of 200 × 109/L prior to storage. Historical
Figure 1.
S59
data in our centre showed no evidence high [WBC] in collected products impacted CD34+ cell recovery or functionality following overnight storage. Therefore, temporary storage of collected products at 4°C without dilution of products to the [WBC] of 200 × 109/L is permited. With the recent updates in collection procedures, we felt that it prudent to re-examine [WBC] influence on CD34+ cell recovery/viability post-cryopreservation and thaw. HPC (A) collections (n = 369) were analyzed to detemine if [WBC] of product post collection impacted cell recovery and viability. All products were cryopreserved the day after collection following overnight storage at 40C. Products were cryopreserved by controlled rate freezer protocol and stored in vapour phase liquid nitrogen freezers until thaw and transplant. Quality Control (QC) vials representative of product were thawed and enumerated for total CD34+ cells surviving and % CD34+ cell viability (7-AAD). Percent (%) CD34+ cell recovery, % viable CD34+ cells, and % WBC recovery were calculated for all products and compared to collected [WBC]. There was no observed difference in % CD34+ cell recovery or viability in the products with Low [WBC] (<200 × 109/L) and High [WBC] (>200 × 109/L). As well, patients receiving products from the “low” and “high” [WBC] groups demonstrated no difference in time to neutrophil or platelet engraftment indicating equivalent functionality of the products. In summary, these findings demonstrate that neither CD34+ cell recovery, viability, nor functionality are impacted by [WBC] higher than 200 × 109/L when HPC (A) products are stored overnight prior to cryopreservation.
122 WILL NOT BE PRESENTED
123 PROLONGED TRANSIT TIME DOES NOT IMPAIR HPC MANIPULATION AND CRYOPRESERVATION P.J. Shaw1,2,3, V. Antonenas1,2, K. Micklethwaite1,2, L.E. Clancy2, K. Yehson1, D. Tong1, D. Gottlieb1,2 1 The Westmead and Children’s Hospital BMT Laboratory, Westmead Hospital, Westmead, New South Wales, Australia, 2Sydney Cellular Therapies Laboratory, Westmead Institute for Medical Research, Westmead, New South Wales, Australia, 3BMT, The Children’s Hospital at Westmead, Sydney, New South Wales, Australia Introduction: In Australia, we commonly obtain cells which have been transported long distances, with significant time delays prior to transplantation, and we have recently reported this. (Patton et al BBMT in press). At The Children’s Hospital, Westmead, it is our standard practice to CD34 select unrelated HPC products ahead of transplant. For this reason, we studied the time taken for the HPC cells to reach our facility and analysed the recovery and purity of CD34 positive cells after selection.