Stem Cell Recovery Following Implementation of an Automated Cord Blood Processing System in a High Volume Laboratory.

Blood ◽  
2007 ◽  
Vol 110 (11) ◽  
pp. 4919-4919
Author(s):  
David T. Harris ◽  
Ingrid E. Paulson ◽  
Heather Brown ◽  
Joseph Rosenthal
Keyword(s):  
Stem Cell ◽  
Cord Blood ◽  
Processing System ◽  
Post Processing ◽  
Cell Recovery ◽  
Recovery Rates ◽  
Cell Dose ◽  
Blood Processing ◽  
Percent Recovery ◽  
The Mean

Abstract Background: Many cord blood (CB) processing facilities are transitioning to automated systems to standardize methods and decrease human error. The AutoXpress Platform™ (AXP™) is an automated, functionally closed, sterile CB processing system. When placed within the AXP device and centrifuged, the whole blood product is separated into its composite cell populations and the TNC fraction of the CB unit is separated and automatically delivered into a blow-molded freezing bag at a uniform volume of 21 mL. Objective: This study evaluated the use of the AXP system in a directed donor (DD) family cord blood bank, targeted at producing consistently high TNC and MNC recovery rates regardless of broad variability in collection volume. Methods: Cord blood units were collected between 12/5/2006 and 2/24/2007 from 1414 consenting mothers who elected to preserve and bank CB at Cord Blood Registry (CBR). Collection kits were provided at the time of enrollment, and after delivery, CB was collected from the umbilical cord and transported to CBR’s processing facility in Tucson, Arizona. During the study period, all units arriving at the CBR laboratory were allocated to either ficoll or AXP processing based on the volume, age, and the degree of clotting in each unit. Units processed using AXP had a volume of 40–130 mL, an age of less than 48 hours since collection, and a clotting score of 0 to 2+ (based on an internal scale). TNC and MNC counts were measured both pre- and post-processing, using the Sysmex analyzer. Results: The mean age of cord blood units arriving at the laboratory was 23.59 hours, and the mean collection volume was 72.93 mL (±18.10mL). The mean TNC count post-processing was 9.94x108, the TNC percent recovery was 96.19%, and the mean MNC percent recovery was 98.65%. Volume (mL) Mean Post-Processing TNC (x108) TNC Percent Recovery (%) Mean Post-Processing MNC (x108) MNC Percent Recovery (%) Mean 72.93 9.94 96.19 4.05 98.65 Standard Deviation 18.10 4.43 11.87 1.59 9.16 Conclusions: AXP automated cord blood processing provides consistently high TNC and MNC recovery rates, which has important implications for stem cell dose if the sample is used in transplant. Because limited cell dose is frequently cited as an obstacle to CB transplantation, processing results could impact the usability of each unit. Because DD family CB banks process all units, regardless of collection volume, percent recovery becomes particularly important in evaluating the differences between processing centers. The AXP system yields the highest published cell recovery rate to date and can be easily integrated into a CB processing center such that it decreases the labor and time required for processing while maintaining MNC recovery of greater than 98%.

scholarly journals TotiCyte, a Paradigm Shift in Stem Cell Isolation and Storage from Umbilical Cord Blood

2021 ◽  
Vol 7 (3) ◽  
pp. 1-7
Author(s):  
Lesley-Ann Martin ◽  
Keyword(s):  
Stem Cell ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Paradigm Shift ◽  
Cell Isolation ◽  
Volume Reduction ◽  
Cell Recovery ◽  
Stem Cell Isolation ◽  
And Storage

We provide evidence to support the use of TotiCyte as a novel volume reduction technology capable of significantly improving CD34+ stem cell recovery

Effect of leukocyte compatibility on neutrophil increment after transfusion of granulocyte colony-stimulating factor–mobilized prophylactic granulocyte transfusions and on clinical outcomes after stem cell transplantation

Blood ◽  
2000 ◽  
Vol 95 (11) ◽  
pp. 3605-3612 ◽  
Author(s):  
Douglas R. Adkins ◽  
Lawrence T. Goodnough ◽  
Shalini Shenoy ◽  
Randy Brown ◽  
Jennifer Moellering ◽  
...  
Keyword(s):  
Stem Cell ◽  
Clinical Outcomes ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Granulocyte Transfusion ◽  
Screening Assay ◽  
Cell Dose ◽  
Component Cell ◽  
The Mean ◽  
Stimulating Factor

The primary limitations of granulocyte transfusions include low component cell dose and leukocyte incompatibility. Component cell dose improved with granulocyte colony-stimulating factor (G-CSF) mobilization, and the transfusion of G-CSF–mobilized, human leukocyte antigen (HLA)-matched granulocyte components resulted in significant, sustained absolute neutrophil count (ANC) increments. However, the effect of leukocyte compatibility on outcomes with G-CSF–mobilized granulocyte transfusions is unclear. The objectives were to determine the effect of leukocyte compatibility on ANC increments and selected clinical outcomes after transfusion of prophylactic, G-CSF–mobilized granulocyte components into neutropenic recipients of autologous peripheral blood stem cell (PBSC) transplants. Beginning on transplant day 2, 23 evaluable recipients were scheduled to receive 4 alternate-day transfusions of granulocyte components apheresed from a single donor given G-CSF. G-CSF was also given to recipients after transplantation. Recipient ANC was determined before and sequentially after each granulocyte transfusion to determine the peak ANC increment. Leukocyte compatibility was determined at study entry only by a lymphocytotoxicity screening assay (s-LCA) against a panel of HLA-defined cells. Eight recipients had positive s-LCA. On days 2 and 4, the mean peak ANC increments after granulocyte transfusion were comparable between the cohorts with positive and negative s-LCA. However, the mean peak ANC increments on day 6 (246/μL vs 724/μL; P = .05) and day 8 (283/μL vs 1079/μL; P = .06) were lower in the cohort with positive s-LCA, in spite of the transfusion of comparable component cell doses. Adverse reactions occurred with only 5 of 87 (5.7%) granulocyte transfusions and were not associated with leukocyte compatibility test results. Platelet increments, determined 1 hour after granulocyte transfusion, were comparable between the cohorts. Although the 2 cohorts received PBSC components with similar CD34+ cell doses, the cohort with a positive s-LCA had delayed neutrophil engraftment and a greater number of febrile days and required more days of intravenous antibiotics and platelet transfusions. Leukocyte incompatibility adversely affected ANC increments after the transfusion of G-CSF–mobilized granulocyte components and clinical outcomes after PBSC transplantation.

scholarly journals ASSESS THE EFFECTIVENESS OF STRUCTURED TEACHING PROGRAMMED REGARDING COLLECTION OF CORD BLOOD FOR STEM CELL THERAPY AT BRAM HOSPITAL OF RAIPUR

2021 ◽  
Vol 9 (09) ◽  
pp. 363-369
Author(s):  
Tripti Goarya ◽  
◽  
Chandrakala Janghel ◽  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cell Therapy ◽  
Cord Blood ◽  
Stem Cell Therapy ◽  
The Body ◽  
Blood Collection ◽  
Structured Teaching ◽  
The Mean ◽  
Cord Blood Collection

The stem cells, derived from the cord blood are hematopoietic stem cells. These have immense potential in curing blood related disorders like blood cancers, thalassemia etc. These can be useful in treating tissue related disorders of heart, bone, spinal cord etc. Stem cells are characterized by the ability to renew through mitotic cell division and differentiate into a diverse range of specialized cell types. Stem Cells are the basic building blocks of the body and have the potential to replenish other cells and give rise to number of tissues which constitute different organs. To conduct the study, 60 antenatal mothers are selected the age range of subjects was 21 to 40 years. The need for the study arises after knowing those antenatal mothers were unaware and having inadequate knowledge regarding cord blood collection for stem cell therapy. The above facts created an interest to conduct a structured teaching programme to assess its effectiveness on knowledge and attitude of antenatal mother on cord blood collection for stem cell therapy. The pretest, the mean score of knowledge is 13.2 mean % is 50.7, & attitude is 31.58, mean % 63.16, the post test the mean score of knowledge is 18.1, mean % is 69.65. Attitude 38.05, mean % 76.1.

Two Cases Report of Haploid Hematopoietic Stem Cell Transplantation Treatment of Adrenal Leukodystrophy

Blood ◽  
2016 ◽  
Vol 128 (22) ◽  
pp. 5878-5878 ◽  
Author(s):  
Libai Chen ◽  
Jianyun Wen ◽  
Yuelin He ◽  
Xiaoqin Feng ◽  
Chunfu Li ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Peripheral Blood ◽  
Peripheral Blood Stem Cell ◽  
Blood Stem Cell ◽  
Hematopoietic Stem ◽  
Cell Dose

Abstract Background : Adrenal leukodystrophy is one of the beta oxidation peroxidase disease, an x-linked recessive heredity, can lead to very long chain fatty acids in tissue accumulation, result in adrenal and cerebral white matter of the progressive deterioration. Hematopoietic stem cell transplantation (HSCT) is a curative treatment for early childhood cerebral type of X-ALD. We report two cases of haploid hematopoietic stem cell transplantation for the treatment of adrenal leukodystrophy. Methods: Two patients were male, 5 years old, 6 years old, respectively, by the gene diagnosis of adrenal leukodystrophy.Case 1 received father haploid bone marrow and peripheral blood stem cell(HLA7/10 match)combine with unrelated umbilical cord blood(HLA 9/10 match). Case 2 received sister haploid bone marrow and peripheral blood stem cell(HLA5/10 match)combine with unrelated umbilical cord blood(HLA 7/10 match).Case 1 conditioning regimen use cyclophosphamide, fludarabine and thiotepa.Case 2 conditioning regimen use cyclophosphamide , busulfan ,fludarabine , rabbit anti-human thymocyte immunoglobulin and thiotepa.Case 1 bone marrow infused total nucleated cell dose was 2×108/kg (CD34+:0.75%,CD3+:1.67%), peripheral blood stem cell infused total nucleated cell dose was 29×108/kg (CD34+:0.19%,CD3+:4.96%), unrelated umbilical cord blood infused total nucleated cell dose was 1.16×108/kg (CD34+:0.44%). Case 2 bone marrow infused total nucleated cell dose was 0.89×108/kg (CD34+:0.74%,CD3+:2.29%), peripheral blood stem cell infused total nucleated cell dose was 25.85×108/kg (CD34+:0.5%,CD3+:15.23%), unrelated umbilical cord blood infused total nucleated cell dose was 0.39×108/kg (CD34+:0.93 %). GVHD prophylaxis Case 1 used mycophenolate mofetil,sirolimus,while Case 2 used mycophenolate mofetil,tacrolimus and sirolimus. Results: The absolute neutrophil count (ANC) greater than 0.5×109/L of two patients were 21 and 23 days , case 1 had a successful engraftment with father donor-derived as case 2 had a successful engraftment with umbilical cord blood donor-derived.Follow-up time of 35 months and 3 months respectively,Case 1 Check head MRI again show a smaller lesionswhile Case 2 progression-free.Two cases' C24:0, C26:0, C24:0 / C22:0, C26:0 / C22:0 of plasma decline than before transplantation. Conclusion: In the absence of HLA-match donor, haploid bone marrow and peripheral blood stem cell combined unrelated umbilical cord transplantation is a effective method of treatment of adrenal leukodystrophy, but which will successful engraftment need more further studies. Disclosures No relevant conflicts of interest to declare.

scholarly journals Comparative Analysis of Cell Dose and Viability of Cord Blood Units at Cryopreservation and at Thaw/Infusion for Unrelated Stem Cell Transplantation in Adult Recipients

2014 ◽  
Vol 20 (2) ◽  
pp. S259-S260
Author(s):  
Spyridoula Vasileiou ◽  
Ioannis Baltadakis ◽  
Fotios Panitsas ◽  
Ifigenia Tzannou ◽  
Zoi Poulopoulou ◽  
...  
Keyword(s):  
Stem Cell ◽  
Comparative Analysis ◽  
Stem Cell Transplantation ◽  
Cord Blood ◽  
Cell Transplantation ◽  
Cell Dose

scholarly journals Safety and feasibility of virus-specific T cells derived from umbilical cord blood in cord blood transplant recipients

2019 ◽  
Vol 3 (14) ◽  
pp. 2057-2068 ◽  
Author(s):  
Allistair A. Abraham ◽  
Tami D. John ◽  
Michael D. Keller ◽  
C. Russell Y. Cruz ◽  
Baheyeldin Salem ◽  
...  
Keyword(s):  
T Cells ◽  
Stem Cell ◽  
Cord Blood ◽  
Ex Vivo ◽  
Cell Receptor ◽  
Viral Disease ◽  
Viral Reactivation ◽  
Cell Transplant ◽  
Transplant Recipients ◽  
Cell Recovery

Abstract Adoptive transfer of virus-specific T cells (VSTs) has been shown to be safe and effective in stem cell transplant recipients. However, the lack of virus-experienced T cells in donor cord blood (CB) has prevented the development of ex vivo expanded donor-derived VSTs for recipients of this stem cell source. Here we evaluated the feasibility and safety of ex vivo expansion of CB T cells from the 20% fraction of the CB unit in pediatric patients receiving a single CB transplant (CBT). In 2 clinical trials conducted at 2 separate sites, we manufactured CB-derived multivirus-specific T cells (CB-VSTs) targeting Epstein-Barr virus (EBV), adenovirus, and cytomegalovirus (CMV) for 18 (86%) of 21 patients demonstrating feasibility. Manufacturing for 2 CB-VSTs failed to meet lot release because of insufficient cell recovery, and there was 1 sterility breach during separation of the frozen 20% fraction. Delayed engraftment was not observed in patients who received the remaining 80% fraction for the primary CBT. There was no grade 3 to 4 acute graft-versus-host disease (GVHD) associated with the infusion of CB-VSTs. None of the 7 patients who received CB-VSTs as prophylaxis developed end-organ disease from CMV, EBV, or adenovirus. In 7 patients receiving CB-VSTs for viral reactivation or infection, only 1 patient developed end-organ viral disease, which was in an immune privileged site (CMV retinitis) and occurred after steroid therapy for GVHD. Finally, we demonstrated the long-term persistence of adoptively transferred CB-VSTs using T-cell receptor-Vβ clonotype tracking, suggesting that CB-VSTs are a feasible addition to antiviral pharmacotherapy.

Effect of leukocyte compatibility on neutrophil increment after transfusion of granulocyte colony-stimulating factor–mobilized prophylactic granulocyte transfusions and on clinical outcomes after stem cell transplantation

Blood ◽  
2000 ◽  
Vol 95 (11) ◽  
pp. 3605-3612 ◽  
Author(s):  
Douglas R. Adkins ◽  
Lawrence T. Goodnough ◽  
Shalini Shenoy ◽  
Randy Brown ◽  
Jennifer Moellering ◽  
...  
Keyword(s):  
Stem Cell ◽  
Clinical Outcomes ◽  
Granulocyte Colony Stimulating Factor ◽  
Colony Stimulating Factor ◽  
Granulocyte Transfusion ◽  
Screening Assay ◽  
Cell Dose ◽  
Component Cell ◽  
The Mean ◽  
Stimulating Factor

Abstract The primary limitations of granulocyte transfusions include low component cell dose and leukocyte incompatibility. Component cell dose improved with granulocyte colony-stimulating factor (G-CSF) mobilization, and the transfusion of G-CSF–mobilized, human leukocyte antigen (HLA)-matched granulocyte components resulted in significant, sustained absolute neutrophil count (ANC) increments. However, the effect of leukocyte compatibility on outcomes with G-CSF–mobilized granulocyte transfusions is unclear. The objectives were to determine the effect of leukocyte compatibility on ANC increments and selected clinical outcomes after transfusion of prophylactic, G-CSF–mobilized granulocyte components into neutropenic recipients of autologous peripheral blood stem cell (PBSC) transplants. Beginning on transplant day 2, 23 evaluable recipients were scheduled to receive 4 alternate-day transfusions of granulocyte components apheresed from a single donor given G-CSF. G-CSF was also given to recipients after transplantation. Recipient ANC was determined before and sequentially after each granulocyte transfusion to determine the peak ANC increment. Leukocyte compatibility was determined at study entry only by a lymphocytotoxicity screening assay (s-LCA) against a panel of HLA-defined cells. Eight recipients had positive s-LCA. On days 2 and 4, the mean peak ANC increments after granulocyte transfusion were comparable between the cohorts with positive and negative s-LCA. However, the mean peak ANC increments on day 6 (246/μL vs 724/μL; P = .05) and day 8 (283/μL vs 1079/μL; P = .06) were lower in the cohort with positive s-LCA, in spite of the transfusion of comparable component cell doses. Adverse reactions occurred with only 5 of 87 (5.7%) granulocyte transfusions and were not associated with leukocyte compatibility test results. Platelet increments, determined 1 hour after granulocyte transfusion, were comparable between the cohorts. Although the 2 cohorts received PBSC components with similar CD34+ cell doses, the cohort with a positive s-LCA had delayed neutrophil engraftment and a greater number of febrile days and required more days of intravenous antibiotics and platelet transfusions. Leukocyte incompatibility adversely affected ANC increments after the transfusion of G-CSF–mobilized granulocyte components and clinical outcomes after PBSC transplantation.

scholarly journals Increasing Post-Processing Total Nucleated Cell (TNC) Recovery in Cord Blood Banking: Hespan Addition in cGMP Environment

Blood ◽  
2014 ◽  
Vol 124 (21) ◽  
pp. 1126-1126
Author(s):  
Ludy Dobrila ◽  
Tracy Zhu ◽  
Dan Zamfir ◽  
Tao Wang ◽  
Michal J Tarnawski ◽  
...  
Keyword(s):  
Cord Blood ◽  
Conflicts Of Interest ◽  
Processing System ◽  
Reduction Process ◽  
Buffy Coat ◽  
Stability Study ◽  
Volume Reduction ◽  
Post Processing ◽  
Hematopoietic Stem ◽  
Cell Counts

Abstract Purpose. Evaluate the effects of HESPAN (HES) addition on indices of cord blood unit (CBU) potency, stability and safety after automated volume reduction (reduction of erythrocyte bulk and plasma volume) using the AXP AutoXpressTM (AXP) processing system. Background and Methods. TNC recovery varies significantly and unpredictably after volume reduction during CBU processing. However, prompt engraftment of CBU allotransplants correlates with their TNC including hematopoietic stem/progenitor cells (HPC). As a result, TNC is important in selecting CBU for transplantation: for example, 75% of the National Cord Blood Program (NCBP) CBUs shipped during the period 2010-2013 had TNC >120 x 107. Therefore, minimizing TNC losses during processing improves the chances that a CBU will be useful for clinical transplantation. The first method to increase post-processing TNC recovery was the addition of HES to the centrifugal method for CB volume reduction [Rubinstein et al, PNAS (USA), 92:10119-10122, 1995]. Volume reduction consists of concentrating the buffy-coat (containing leukocytes and HPC) by centrifugal stratification and removal of bulk red cells and platelet-containing plasma. NCBP processed manually over 30,000 CBUs using HES during the period 1995-2006. In 2006, the AXP, a closed, automated and FDA-approved processing method, without HES, was implemented. AXP was designed to enhance the MNC and CD34+ cell recoveries but not those of granulocytes. This resulted in lower post- processing TNC counts. Additionally, CBUs with larger volume and TNC content have somewhat higher TNC losses during automated processing. Using the AXP method, NCBP’s HEMACORD® obtained FDA License approval on November 10, 2011. However, since TNC (not MNC or CD34+ count) remains the most commonly used indicator of CBU potency and engraftment ability, we describe here implementation of HES in AXP processing to augment TNC recovery, by adding HES to the CBU to a 1-2% final concentration. Results.The results of manual CBU processing showed that HES addition improves yield, without changes in cell viability after cryopreservation, freezing at -196°C and thawing, after 20 years (from NCBP continuing stability study). HES addition also preserved CFU numbers and CD34+ cell counts after thawing. 1. Comparison of TNC recoveries without and with HES addition in the same CBU: A total of 25 CBUs were initially processed with the AXP platform without HES, as per routine procedure, and the TNC recoveries were calculated. Each CBU was then reconstituted after its initial processing into a new AXP bag set. HES was added aseptically to the reconstituted product and each CBU was processed in the same conditions as first time (same AXP device, centrifuge, etc.). The TNC recoveries after the second volume reduction process (AXP with HES) were ~20% higher on average than after the first (AXP without HES). 2. Comparison of TNC recoveries in different cohorts of CBUs: Thirty clinical-grade CBUs, with volumes 80-156 mL and TNC counts 111-290 x 107, were processed with HES in the AXP system and the results were compared with those of AXP-processed CBUs without HES over an earlier six month period. TNC and CD45+ cell recoveries improved by 16 - 20% maintaining mean post-processing hematocrit at 30.6% (SD ± 2). CD34+ and CD45+ cell viabilities were unchanged: 99% (SD ± 0.7) and 96% (SD ± 2.7), respectively, while their mean recoveries were 95% (SD ± 18) and 94% (SD ± 6). In addition, the same consistent post-processing volume was obtained (mean 20.8 mL, SD ± 0.1). CFU - CD34+ correlation (R2) after processing with HES was 0.788 (not different than what was observed in the CBUs without HES). Six CBUs AXP-processed with HES, were thawed and tested, with very minor losses in cell count and viability, similar to results of thawed AXP-processed CBU without HES. Finally, HES addition did not result in microbial contamination in any of these AXP-processed CBUs. Conclusion. Adding HES to CBUs before automated AXP processing increased substantially the TNC and CD45+ recoveries without loss of viability, while the CD34+ recoveries remained basically unchanged, with a mean of 95%. The post-processing hematocrit was consistent and low. AXP-automated CB processing with HES addition can be performed in the GMP environment, results in higher post-processing TNC and therefore, increases the CB bank’s ability to store larger CBUs that are most useful to patients. Disclosures No relevant conflicts of interest to declare.

scholarly journals Allogeneic stem cell transplantation with omidubicel in sickle cell disease

2021 ◽  
Vol 5 (3) ◽  
pp. 843-852
Author(s):  
Suhag Parikh ◽  
Joel A. Brochstein ◽  
Einat Galamidi ◽  
Aurélie Schwarzbach ◽  
Joanne Kurtzberg
Keyword(s):  
Stem Cell ◽  
Sickle Cell Disease ◽  
Stem Cell Transplantation ◽  
Cord Blood ◽  
Cell Transplantation ◽  
Sickle Cell ◽  
Ex Vivo ◽  
Cell Disease ◽  
Hematopoietic Stem ◽  
Cell Dose

Abstract Many patients with sickle cell disease (SCD) do not have HLA-matched related donors for hematopoietic stem cell transplantation (HSCT). Unrelated cord blood (UCB) is an alternative graft option but is historically associated with high graft failure rates, with inadequate cell dose a major limitation. Omidubicel is a nicotinamide-based, ex vivo–expanded UCB product associated with rapid engraftment in adults with hematologic malignancies. We hypothesized that increasing the UCB cell dose with this strategy would lead to improved engraftment in pediatric patients undergoing myeloablative HSCT for SCD. We report the outcomes of a phase 1/2 study in 13 patients with severe SCD who received omidubicel in combination with an unmanipulated UCB graft and 3 who received a single omidubicel graft. Grafts were minimally matched with patients at 4 of 6 HLA alleles. Median age at transplant was 13 years. A median CD34+ expansion of ∼80-fold was observed in omidubicel and led to rapid neutrophil engraftment (median, 7 days). Long-term engraftment was derived from the unmanipulated graft in most of the double cord blood recipients. Two of the 3 single omidubicel recipients also had sustained engraftment. Incidence of acute graft-versus-host disease (GVHD) was high, but resolved in all surviving patients. Event-free survival in the double cord group was 85% (median follow-up 4 years). All 3 patients in the single cord group were alive at 1 year after transplantation. Ex vivo expansion of UCB with omidubicel supports engraftment in patients with SCD. This approach to decreasing the incidence of GVHD should be optimized for general use in patients with SCD. This study was registered at www.clinicaltrials.gov as #NCT01590628.

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