Cord Blood Units Research Articles

Abstract 26 HLA Analysis of the Australian Cord Blood Banks: How Diverse Are Donors?

IntroductionThe network of public cord blood banks (CBB) in Australia, known as AusCord, is comprised of CBB located in Brisbane, Sydney, and Melbourne. The network stores almost 37,000 cord blood units (CBU) and has released more than 1,300 for transplantation.ObjectiveThe objectives of this study were (1) to determine the relative diversity of HLA allele subtypes, tissue types, and haplotypes at each of the banks and between the banks and (2) to identify common tissue types and haplotypes that could be utilized for clinical research and development of third party cell therapy products.MethodsHLA data was obtained for 36,782 CBU stored in the AusCord inventory. To standardize data format, high-resolution typing was converted to 2-digit typing. HLA allele subtypes were ranked from most to least common. A subset of Indigenous Australian and Pacific Islander HLA allele subtypes was interrogated to determine whether increased frequency correlated with declared ethnicity. Tissue types were separated into CBU that had HLA-A, HLA-B, HLA-C, and HLA-DRB1 typing (21,815 total) and CBU that did not have HLA-C typing (36,782 total). CBU tissue types were interrogated in 3 separate groups: searchable, searched, and released inventory. Haplotypes were confirmed where maternal typing was available (3,105 CBU).ResultsEthnicity screening for donors and strategic location of collection sites servicing ethnic minority communities resulted in banking of Indigenous Australian and Pacific Islander HLA. At a bank and network level, there was a similar frequency of specific HLA subtypes; however, when considering the tissue types, there was vast diversity (~75% unique with 8 allele typing). Whilst there is diversity, there was also a fraction of the inventory that exhibited repetitive tissue types that could be utilized for clinical research.DiscussionThe study demonstrates that cord blood collections are able to boost storage of diverse tissue types in, and unique to, a multicultural population such as Australia. This finding can guide policy development and funding, as well as operational decisions at the CBB. Furthermore, CBB are well positioned to support research and development activity aimed at discovery of new applications for cord blood units through supply of GMP grade cryopreserved products exhibiting common tissue types and haplotypes.

Abstract 23 A First Insight into the Validity of Post-Thaw Cell Recovery, Viability, and Potency Over Time

IntroductionTesting of a thawed segment or representative sample of a cord blood unit (CBU) for cell recovery, viability, and potency forms part of pre-release testing. Not all CBUs that have had pre-release testing completed are shipped for transplant, due to postponed or cancelled shipment request, and are therefore kept in the inventory until they are searched again. In an effort to identify these CBUs for future expedited release, the World Marrow Donor Association (WMDA) initiated a project to flag them as “Ready to Ship” in the WMDA Search & Match Service. However, it is not known how long post-thaw results are valid before they must be repeated on another thaw. Such information can be beneficial not only for transplant centers to better assess CBU suitability but also for Cord Blood Banks (CBBs) to avoid unnecessarily exhausting their precious samples.ObjectiveThis study aimed to provide a first insight into the validity of post-thaw results over various lengths of time.MethodsNineteen CBUs, collected between 2003 and 2015 at the BMDI CBB and that had post-thaw testing performed twice, were analyzed for change in the total nucleated cell (TNC) recovery, total CD34 cell recovery, CD34 cell viability, and CFU-GM counts between two thaws.ResultsThe 19 CBUs were grouped into 4 groups based on their storage time between thaws. An average decrease in TNC recovery of 5.3% from first thaw to second thaw was observed for the entire dataset (P = .015). Four CBUs with a greater than 10% decrease had a bag TNC greater than 25 × 108. Individual CBU data is shown in Figure 1. No significant change between thaws was seen with total CD34 recovery (P = .738) or CD34 viability (n = 9 pairs, P = .354). A large variation in CFU-GM counts was observed between thaws. It is worth noting that all CBUs met CBB release criteria of >60% TNC recovery and that CD34 viabilities were above 70% (FACT requirement) in both thaws.Figure 1.Comparison of first and second thaw TNC recovery (A), total CD34 recovery (B), CD34 viability (C), and CFU-GM counts (D) of individual cord blood unit, grouped into 4 groups based on their storage time between thaws: 0-6 months (i), 2-3.5 years (ii), 6-8.5 years (iii), and 12 years (iv).DiscussionWhile results need to be confirmed on a larger dataset, our preliminary data shows very little change in post-thaw cell recovery and viability over various lengths of time (2 months-12 years), suggesting post-thaw results may be valid for longer than 10 years.

Abstract 33 A Diverse and Sustainable State-Run Public Cord Blood Program

IntroductionThe Arizona Public Cord Blood Program was created in 2011 by the Arizona Biomedical Research Centre, a subsection of the Arizona Department of Health Services, to advance the collection and increase the number of racially and ethnically diverse cord blood units available for transplantation, as well as to promote awareness of the benefits of cord blood stem cells through our educational partner, Save the Cord Foundation (STCF).ObjectiveThe main objective of this study was to create a sustainable program for women of all racial and ethnic backgrounds to have the opportunity to donate cord blood, with the primary goal of transplantation and the secondary goal of providing non-transplantable cord blood units for research. A second objective was to educate the residents of the state of Arizona about cord blood stem cells and the need for their preservation.MethodsA portion of state lottery funds support the program monetarily. Those funds are provided to four partner collection hospitals employing "cord blood consenters," whose responsibility it is to consent patients, assist delivery providers with collections, and package and ship cord blood units to our partner cord blood bank at MD Anderson Cancer Center. There are also two clinical coordinators who educate and train hospital staff on quality collection practices, with special emphasis on the importance of high-volume, sterile collections. STCF provides education across the state to expectant parents, health care providers, schools, and the public about the need for cord blood stem cell donation for transplant and research.ResultsSince 2011, the Arizona Public Cord Blood Program has banked several hundreds of racially and ethnically diverse cord blood units with the National Marrow Donor Program (Figure 1) and has had 80 life-saving cord blood units matched with patients in need around the globe. This innovative program has expanded cord blood awareness and promoted the preservation of cord blood, and it also has resulted in the creation of an economic engine for the state of Arizona that is an attractant for STEM-based businesses and careers.Figure 1.Pie chart showing percentage of collections by race/ethnicity. Abbreviation: AI/AN, American Indian/Alaskan Native.DiscussionA decade later, the Arizona Public Cord Blood Program has proven to be a sustainable model for collecting and providing suitable cord blood units for transplant to diverse patient populations.

Abstract 32 Low Incidence of Product Complaints and Adverse Events Support Safety of Cord Blood Units

IntroductionAs an important cellular therapy source for many patients, the safe and timely delivery of cord blood units (CBU) is paramount. The National Marrow Donor Program (NMDP) investigates product complaints received for CBUs distributed under the 10-CBA investigational new drug application that may impact the safety, quality, identity, purity, or potency of the product. NMDP also receives reports of any patient adverse events or serious complications resulting from the transplantation of CBUs. Information on product complaints and adverse events provides insight into areas related to product integrity, patient safety, and customer satisfaction and highlights opportunities for improvement.ObjectiveWe analyzed all product complaints and adverse events reports submitted to NMDP for CBUs distributed under the NMDP 2010 Cord Blood Access Protocol (10-CBA) from October 2019 through March 2022. These investigations offer visibility into factors associated with product quality and integrity discovered after shipment.MethodsA total of 1,578 CBUs from 38 cord blood banks were shipped from October 2019 through March 2022 (30 months). Data on CBU product complaints reported to NMDP were compiled and analyzed. Data were grouped into categories based upon the primary reason for the product complaint or adverse event. The outcome of NMDP investigations and resolutions is relayed quarterly to the Quality Subgroup of the Cord Blood Advisory Group.ResultsOf the 1,578 CBUs shipped between October 2019 and March 2022, 98% had no reported incidents (32 reports of product complaints and 4 of adverse events). The most common reasons for product complaints investigated by NMDP were related to shipping/transportation (n = 15, 47%), quality issues such as lower than expected potency or bag breaks (n = 11, 34%), and labeling errors (n = 3, 9%). Three of the four adverse events were associated with infusion (two hypertensive episodes and one anaphylaxis) and one was a secondary cancer following transplant.DiscussionTransplant centers report very few quality incidents, product complaints, or adverse events when using cord blood units. We remain vigilant for opportunities to improve. Clinicians and patients can be assured that CBUs are a safe product with high likelihood of arriving to the transplant center without incident or causing adverse events once transplanted.

Abstract 28 Comparative Analysis of the Cord Blood Bank in Quebec

IntroductionIn the context of the regional diversity observed in the population of Quebec, the Héma-Québec Cord Blood Bank (HQ CBB) representation differs from that of the Héma-Québec Stem Cell Registry (HQ Registry). The Bank encourages registrants of non-European descent to promote diversity, and only collects from health centers around Montreal and Quebec cities because of proximity concerns.ObjectiveIn this study, we aimed to analyze allele frequencies in the HQ CBB to assess similarities and differences in reference to the HQ Registry.MethodsThe HQ Registry was previously typed using high-resolution genotyping. For the HQ CBB, ambiguous two-field HLA-A, -B, -C, and -DRB1 typing data were obtained from cord blood units. Statistical analyses were performed using the GENE[RATE] population analysis tools to estimate allele and haplotype frequencies by administrative region determined by registered postal codes. Hardy-Weinberg equilibrium was verified by the nested likelihood procedure. Finally, genetic distance was compared between the HQ Registry and the HQ CBB by region using Non-metric Multi-dimensional Scaling (NMDS).ResultsBecause of the small sample size in certain regions, HLA frequency was analyzed for 7 of the 17 regions within the HQ CBB with typings from 11,472 cord blood units. The HLA allele frequencies were previously assessed for the Registry, with 3,806 donors originating from 14 administrative regions. The Hardy-Weinberg equilibrium was rejected at least in relation to one locus in 3 regions (Capitale-Nationale, Montreal, and Laval), which differs from the results obtained from the Registry, in which the equilibrium was rejected only for Montreal. As in the Registry, allele frequencies in the HQ CBB varied by region but had limited overlap with the HQ Registry. Two regions (Chaudière-Appalaches and Laval) demonstrated significant differences in allele frequencies as represented in the HQ Registry versus the HQ CBB.DiscussionWe observed differences in the Quebec population representation between the HQ CBB and the HQ Registry. This probably represents selection bias related to recruitment strategies and inclusion criteria. Importantly, the complementarity of these two stem cell sources could allow for a more diverse donor pool, and this understanding could also guide the recruitment strategies.

Abstract 19 Cleveland Cord Blood Center (CCBC) Best Practices Throughout the COVID-19 Pandemic and How It Impacted Collection of Cord Blood Units

IntroductionOur commitment to provide quality cord blood units (CBUs) to patients in need was challenged with the outbreak of COVID-19 and the pandemics continued impact through early 2022. We continually monitored the ever-evolving COVID-19 public health threat to make adjustments to our practices to achieve optimal collections while maintaining the safety and quality of our CBUs.ObjectiveThe CCBC continued collections at all five collection sites with a brief mandated pause at our California site due to hospital restrictions. We maintained minimal staffing at the cord blood center to reduce the number of employees impacted should an exposure occur. Donors were screened upon admission for symptoms, exposure, positive testing, and recent vaccination, and donors with identified risks were excluded from collection.MethodsOur study design takes a retrospective look at data collected from the onset of the COVID-19 pandemic through December 2021 compared to data from 2018-2019. Factors included were birth numbers, number of patients testing positive for COVID-19, and staffing shortages due to coordinator illness with COVID-19.ResultsWe saw limited impact to our collection staff at the hospital or on site at the CCBC. Our biggest reduction of donors consented was between 2019 and 2020, which we attribute to the COVID peak and introduction of vaccinations. Most sites were trending downward for consenting on a yearly basis before COVID-19 at a rate of 6%-10% per year; during 2019-2020 we saw 19%, but the trend has returned to 8% from 2020-2021. This same trending occurred during the ZIKA outbreak. There was no further data review to determine if these mothers would have been excluded for other reasons. We collected a total of 15,561 units between 2018 and 2019 and a total of 10,121 units between 2020 and 2021.DiscussionOverall productivity of the collections was not dramatically affected by our response to the COVID-19 pandemic. Further research at time of pre-consent, to determine if the mother would have been ineligible to donate for reasons other than COVID-19, would have given us a more accurate count of donors lost for reasons related to the COVID-19 pandemic.

Abstract 25 ECTmatch: Optimizing Small-Scale Cord Blood Banking Through HLA Analysis

IntroductionCord blood (CB) banks have had to rely on large inventories of CB units to try to serve the largest possible proportion of the population, all the while prioritizing collection of non-Caucasian ethnic groups. However, due to the high linkage disequilibrium of HLA genes and the high frequency of several HLA alleles in the population, CB banks contain hundreds of CB units that could be matched to the same patients, making the inventory somewhat redundant from a clinical standpoint.ObjectiveExCellThera developed ECTmatch, an algorithm dedicated to optimizing the selection of CB units based on in-depth HLA analysis in order to maximize the efficiency of the bank to suitably match the largest proportion of subjects within a small pool of donors.MethodsThe performance of ECTmatch was evaluated in a simulation aiming to select 100 CB units from the Héma-Québec CB bank that satisfied an arbitrary minimal cell content criteria of 120 × 107 TNC and 6 × 106 CD34+ cells (n = 2,987). Selection was performed to optimize matching for the Quebec population, with a minimal HLA-match of 5/8 for HLA-A, -B, -C, and -DRB1.ResultsECTmatch provides a suitably matched donor for 71.5% of the Quebec population, compared with only 45.0% (±2.4%) with random selection. Because patients who require a CB transplant tend to have rarer HLAs, the performance of ECTmatch was evaluated for this specific subset of patients (n = 62). Again, ECTmatch outperformed random selection, by providing a donor for 54.8% of patients, compared with only 29.9% with random selection. Finally, while ECTmatch was developed to optimize CB selection specifically for the Quebec population, it still outperformed random selection for subjects from the other Canadian provinces or the USA.DiscussionBy selecting CB units based on HLA profiles, ECTmatch allows the creation of a highly useful inventory with a very low number of CB units. This approach to small-scale CB banking can be adapted to different population subsets and could be used to select a subset of CB units for pre-release for immediate clinical availability or for the creation of a pre-expanded CB inventory with maximal population coverage.

Abstract 13 A Randomized, Placebo-Controlled, Phase II Trial of Intravenous Allogeneic Umbilical Cord Blood Infusion for Adults with Ischemic Stroke

IntroductionStroke is the fifth leading cause of death in the United States. Tissue plasminogen activator and mechanical thrombectomy are the only effective treatments, but many patients are ineligible for these treatments.ObjectiveThe objective of this study was to determine whether an intravenous infusion of a non-HLA matched, unrelated donor umbilical cord blood (UCB) would improve functional outcomes.MethodsWe conducted a phase II multicenter, randomized (2:1), placebo controlled, double-blinded trial of UCB in adults with acute ischemic stroke. Patients had to have adequate immune function. Cord blood units were selected from U.S. public cord banks based on blood type, race, and cell dose. Study product was infused 3-10 days post stroke. Participants were randomized within strata of National Institutes of Health Stroke Scale Score (NIHSS) (<12 vs ≥12), and study center. The primary endpoint was change in Modified Rankin Scale (mRS) (baseline minus day 90). The study was powered at 80% (odds ratio of 2). Key secondary outcomes included functional independence at day 90 (mRS <2), NIHSS, the Barthel Index, infusion reactions, and adverse events.ResultsSeventy-nine participants were enrolled at 6 centers when the trial was closed early due to slow accrual related to COVID19; 73 participants (47 randomized to UCB) were included in the safety and efficacy analyses. The median (range) of the change in mRS was 1 (–2, 3) in UCB and 1 (–1, 4) in placebo. A shift analysis based on the proportional odds model showed an odds ratio of 0.9 (95% CI: 0.4, 2.3) after adjustment for baseline mRS and randomization strata. No differences were observed on the key secondary outcomes. There were 17 mild infusion reactions (27.6% UCB; 15.4% placebo). The distribution of serious and non-serious adverse events was similar between arms.DiscussionThis study demonstrated the safety of infusing non-HLA matched UCB to adults with acute ischemic stroke. Feasibility and logistics were challenging. The primary efficacy endpoint did not demonstrate benefit in this underpowered sample size. In a secondary ad hoc analysis, a trend of improved functional outcomes at day 90 in recipients of UCB more than 5 days post stroke (Figure 1) could be explored in future trials.Figure 1.Cord blood versus placebo odds ratio by treatment latency (days).

Abstract 7 Reduced Intensity Conditioning Umbilical Cord Blood Transplantation in Combination with Splenic Irradiation in a Patient with Secondary Myelofibrosis and Massive Splenomegaly

IntroductionMyelofibrosis (MF) is a bone marrow neoplasm characterized by marrow fibrosis/failure and splenomegaly, causing pain, early satiety, and cytopenia. The only curative option is allogeneic hematopoietic stem cell transplant (allo-HSCT); however, its use is limited due to donor availability and patient age, functional status, and comorbidities. Umbilical cord blood (UCB) has been explored in this population for this reason with variable success, partly related to splenomegaly, which might influence engraftment in MF patients.ObjectiveThe objective was to report the successful use of UCB transplantation in conjunction with splenic irradiation to treat a patient with advanced MF and massive splenomegaly.MethodsA 71-year-old male with post-essential thrombocythemia MF was treated with ruxolitinib for 2 years but developed worsening cytopenia and transfusion dependence. Splenomegaly (19.1×7.1×20.8 cm) was identified. Allo-HSCT was recommended on the basis of high-risk disease (DIPSS Plus score: 4) and few alternative options. Although suitable HLA-matched donors were unattainable, two cord blood units were available. A reduced intensity conditioning regimen of fludarabine, cyclophosphamide, and TBI 300 was chosen. Given the primary resistance to ruxolitinib, the spleen was irradiated with 450 cGy prior to transplant.ResultsNeutrophil and platelet engraftment occurred on days +30 and +49, respectively. The patient developed engraftment syndrome and acute graft versus host disease (GVHD) that resolved with methylprednisolone and early introduction of ruxolitinib. His immunosuppressive medications were discontinued by 6-months without evidence of chronic GVHD thereafter. He is now 1 year post-transplant with 100% donor chimerism and no clinical evidence of MF or GVHD.DiscussionWe present a novel therapeutic approach combining the use of splenic irradiation and a double unit UCB HSCT in a patient with MF and massive splenomegaly. UCB transplant should be considered in patients with MF who do not have an HLA-matched donor with the additional benefit of less chronic GvHD. Massive splenomegaly can negatively impact transplant outcomes due to entrapment of donor stem cells, delayed engraftment, and prolonged cytopenia. Splenic irradiation before UCB transplant can potentially reduce these complications, as it did in this case, and in turn decrease transplant-related mortality. Our future aim is to develop a clinical trial to study this approach.

Abstract 24 Improving Cord Blood Unit Selection and Optimizing Manufacturing of a Unique Cord Blood-Derived Product (DUOC-01)

IntroductionDUOC-01 are microglia-like cells derived from umbilical cord blood CD14-positive monocytes, which have been shown to induce remyelination in murine models of demyelination. Intrathecal DUOC-01 administration is being tested clinically in two phase I studies. First, DUOC-01 is being tested as a bridging therapy for pediatric patients with inherited leukodystrophies undergoing unrelated donor cord blood transplantation. Second, DUOC-01 is being tested in a phase Ia dose escalation study in patients with primary progressive multiple sclerosis.ObjectiveDUOC-01 is manufactured from an HLA-matched cryopreserved cord blood unit (CBU). The CBU is thawed and cultured for 21 days under specific conditions, and the resulting DUOC-01 is harvested and formulated with hydrocortisone for intrathecal administration. Cell dosing per patient is limited by DUOC-01 yield, which is highly variable; therefore, there is great need to increase DUOC-01 yield and optimize manufacturing for more consistency.Methods(1) For the identification of optimal CBU selection, units are screened using small scale DUOC-01 cultures derived from CBU bag segments, which are then scored (0-4). In order to assess the value, we performed screening followed by DUOC-01 manufacturing and correlated the screening score to the DUOC-01 yield. (2) We assessed multiple variables to optimize DUOC-01 manufacturing including media volume and temperature, fetal bovine serum (FBS) lot variability, and culture container material.Results(1) 5/13 segments scored high (defined as ≥2) while the remaining 8/13 scored low (<2). All 5 high-scored CBUs resulted in high yield (defined as ≥1.64 total cells/cm2). Of the 8 low-scored segments, 4 were low yield and 4 were high yield. (2) Comparison of different FBS lots did not show a significant change in DUOC-01 yield. Furthermore, media volume and temperature did not influence product yield. The choice of container material substantially impacted culture yields and cell viability (Figure 1).Figure 1Choice of tissue culture flasks affects DUOC-01 cell yields and viabilities. (A): Average live yield comparison of harvested cells. (B): Average cell viability comparison of harvested cells. Each color/symbol represents the same source umbilical cord blood (paired, n varies from 1 to 4). Bars show means. Statistical analysis was performed by paired T test.DiscussionIn order to increase cell yield and improve product consistency, we confirmed the value of the CBU segment screening assay and discovered the potential importance of culture container material. More experiments are needed to optimize and finalize product manufacturing.

Abstract 21 High-Throughput Blood Separation System for CD34+ Cell Isolation

IntroductionResearch into the therapeutic roles and clinical potential of CD34+ cells is growing [Weissman and Shizuru, Blood. 2008;112(9):3543-3553]. Accordingly, advances in cell isolation methods have risen to meet the research demand. Cord blood banks have a unique opportunity to source CD34+ cells because of their relatively high prevalence in cord blood [Dauber et al. Cytotherapy. 2011;13(4):449-458]. However, current isolation systems are not optimized for processing cord blood, and even blood-compatible systems typically require the blood to be processed before use. FerroBio (www.42bio.com) has developed a magnetic separation system that facilitates scalable, high-volume cell isolation from complex biological fluids, such as blood, without pre-processing. This technology is now being adapted for CD34+ cell isolation from cord blood.ObjectiveIn this study, the conditions in which magnetic beads efficiently bind target cells were explored. The objective of this work was to evaluate optimal blood mixing conditions to bring beads into contact with cells in suspension in the blood. This will allow unmodified cord blood units (CBUs) to flow directly into our apparatus.MethodsThe FerroBio separation process is comprised of three simple steps (Figure 1A). Here, conditions of step 1 were explored to maximize target cell recovery and viability. Variables included type of agitation equipment (Figure 1B), temperature, time, and relative air volume in the blood bag. Recovery and viability were measured via flow cytometry and calculated relative to unprocessed blood from the same CBU.ResultsTwo mixing conditions were identified for maximum recovery of CD34+ cells (Figure 1C, blue bars). While not statistically different from each other, the Drum Roller method only requires one piece of standard equipment, is gentler, and can accommodate multiple CBUs simultaneously. No statistical difference in viability was observed in any conditions (90.7% ± 16.3%, n = 9). Increasing incubation time and the relative amount of air in the bag did not increase recovery (not shown).Figure 1. (A): Overview of the FerroBio Magnetic Separation Process, highlighting step (1) in which magnetic beads are mixed within the CBU to bind the target CD34+ cells. (B): Illustrations of the agitation equipment used to mix the CBU during step (1). (C): The resulting CD34+ recovery (1-way ANOVA, P = .0245). Image credit for panel (A): BioRender.DiscussionThe FerroBio separator can streamline CD34+ cell isolation by processing high blood volumes in a fast, closed system with high recovery rates. Additional studies are under way to evaluate the functionality of the isolated CD34+ cells and efficiency of bead removal.

Abstract 11 Outcome of Patients Treated with Umbilical Cord Blood and Homologous Serum Eye Drops

IntroductionSeveral authors have demonstrated the safety and efficacy of CBS eye drops in the treatment of severe DE with or without Sjogren’s syndrome (SS-I), ocular graft versus host disease (GVHD), persistent corneal epithelial defects, recurrent corneal erosions, chemical burns, and neurotrophic keratitis.ObjectiveTo compare the efficacy of cord blood serum (CBS) and adult donor peripheral blood serum (PBS) eye drops (prepared, standardized, and controlled in the Emilia Romagna Cord Blood Bank) in the treatment of severe dry eye disease (DED), a multicenter randomized, double masked, crossover clinical trial was designed.MethodsSixty patients diagnosed with severe DED associated with persistent corneal epithelial defects were randomized and equally assigned to group A (treated with CBS) or group B (treated with PBS) , eye drops administered 8 times/day for 1 month. Primary outcome was the pre-treatment and post-treatment change in corneal fluorescein staining. Secondary outcomes included the pretreatment and post-treatment change in ocular surface disease index (OSDI), questionnaire and Visual Analogue Score (VAS) of subjective symptoms, Schirmer I test, tear break-up time, and conjunctival staining. Patients with relapse in signs or symptoms after further 2 months switched to the remaining group for one additional month.ResultsCorneal staining was more significantly reduced after CBS treatment; both VAS and OSDI score reduction was observed in both groups, but group A reported significantly less grittiness and pain. Nineteen patients shifted in the crossover period. The within individual comparison confirmed a better recovery in the CBS treatment period. Epidermal growth factor, transforming growth factor, and platelet-derived growth factor content was associated with reduction of epithelial damage. Levels of interleukins were positively associated with symptoms decrease.DiscussionDED signs improved after both CBS and PBS treatments with potential advantages of CBS for subjective symptoms and corneal damage reduction. In addition to the very important therapeutic value of EDHO products, considering the CB units not suitable for transplantation, we have identified novel non-wasteful clinical applications of those allogeneic biological products, resulting in an increase of ethical motivation of families generously donating CB for public benefit.Figure 1.Slit-lamp photographs with fluorescein staining in patient running both period 1 and period 2 of the study. Before PBS treatment, the corneal epithelium was stained diffusely (top left). One month after treatment, the severity of the damage decreased, especially in the central area (top right). Two months after the end of the treatment, the level of epithelial damage worsened and patient entered period 2 (bottom left). After 1 month of CBS treatment, a diffuse decrease in staining was observed in all areas. In this patient, a better improvement was observed after the CBS treatment compared with the PBS treatment.

Abstract 15 Safety of DUOC-01, Intrathecal Cord Blood-Derived Cellular Therapy, as an Adjunct to Allogeneic Cord Blood Transplantation in Children with Inherited Metabolic Diseases

IntroductionUnrelated donor umbilical cord blood transplantation (CBT) improves function and extends life in children with certain inherited metabolic diseases (IMDs); however, progressive neurologic decline often occurs during the few months post-CBT prior to donor engraftment in the brain, causing residual, irreversible impairments. We previously described donor engraftment in the brain after CBT and developed DUOC-01, a monocyte-derived CB cell product, to attempt to accelerate donor cell delivery to the brain.ObjectiveThe objective of this study was to determine manufacturing feasibility and safety of intrathecal DUOC-01 in a phase I study in children with IMDs undergoing CBT after myeloablative cytoreduction (busulfan/cyclophosphamide/anti-thymocyte globulin).MethodsDUOC-01 was manufactured in adherent cell culture in a GMP lab over 21 days with a target dose of ≤100 × 105 cells. When cell dose permitted, 80% of the CB unit was used for transplant and 20% of the same unit was used to manufacture DUOC-01. Otherwise, DUOC-01 was manufactured from a unique CB unit. On day ≥+28 post-CBT, if there was evidence of hematopoietic engraftment without severe graft versus host disease or morbidity, DUOC-01 cells were harvested and administered intrathecally. Safety was monitored via neurologic exams, neuroimaging, and CSF studies.ResultsForty patients (aged 0-15 years) underwent CBT, 37 engrafted with donor cells. Median infused total nucleated cell count was 6.5 × 107/kg (range 2.9-33.1 × 107/kg); median time to neutrophil engraftment was 20 days (range 13-46 days). Ten patients did not receive DUOC-01 cells for the following reasons: engraftment failure (3), transplant-related morbidity (4), and low manufacturing yield (3). Thirty patients received 0.2-100 × 105 DUOC-01 cells intrathecally for a median of 31 days (range 27-77 days) post-CBT (19 with the same CB donor and 11 with a unique CB donor). Two patients developed transient fever and hypotension hours after unique-donor DUOC-01, consistent with an inflammatory response. Hydrocortisone (HC) was added to the final formulation, and post-validation, seven additional patients received unique-donor DUOC-01+HC without incident. No other DUOC-01-related serious adverse events occurred.DiscussionDUOC-01 is a CB-derived product intended to accelerate donor cell delivery to the brain of children with IMDs undergoing CBT. Intrathecal DUOC-01 administration is well-tolerated post-CBT from the same donor and, with the addition of HC, also from a unique donor. Escalating DUOC doses are being tested for safety and exploration of potential efficacy in a phase Ia trial of adults with multiple sclerosis.

Abstract 16 New Advanced Therapy Medicinal Products (ATMPs) in Medical Trials: A FamiCord Group Development Pipeline Update

IntroductionSeventeen products in five countries are used in medical experiments and clinical trials. FamiCord Group, the leading stem cell bank in Europe and number 3 worldwide, is a listed biotech group consisting of the European biotech companies Vita 34 (Germany), P.B.K.M. (Poland), Crioestaminal (Portugal), Smart Cells (UK), and more than 30 other subsidiaries. FamiCord Group was founded in 2002 and is located in more than 35 countries in Europe and the Middle East. The Group is engaged in the storage of biological material and the manufacture, development, and approval of cell therapeutics for clinical use. More than 850,000 cryopreserved umbilical cord blood and tissue units and more than 4,200 patients treated with stem cells in clinical trials, experimental therapies, and standard therapies demonstrate the extraordinary expertise in stem cell banking and clinical application.ObjectiveAn overview of the Cell Therapy Pipeline with cell products from the FamiCord Group is shown in Table 1. ATMPs have been developed and manufactured in 3 synergistic R&D centers in Poland, Portugal, and Germany. Twenty preclinical and clinical trials are being conducted as part of in-house and consortia projects.Table 1.Overview of the Cell Therapy Pipeline using cell products of FamiCord GroupCountryProject titlePhaseIndicationGermanyCartiCureClinical trial phase I/IIaCartilage damageGermanyArthroCurePreclinical trial/PoCOsteoarthritisSwedenPro-Trans-1Clinical trial phase I/IIDiabetes type ISwedenPro-Trans-RepeatClinical trial phase IIDiabetes type ISwedenProTrans-YoungClinical trial phase I/IIDiabetes type ISwedenProTrans-19+SEClinical trial phase ICOVID-19CanadaProTrans-19+CAClinical trial phase IICOVID-19PortugalSTROKE34Clinical trialStrokePortugalRescueCordHospital exemptionHIEPortugalMSC-ARDSClinical trial phase IIARDSPortugalMSC-GvHDHospital exemptionAcute GvHDPolandALSTEMClinical trial phase I/IIaALSPolandBIOOPAClinical trial phase I/IIEpidermolysis bullosaPolandCIRCULATEClinical trial phase II/IIINo-option critical limb ischemiaPolandCIRCULATEClinical trial phase II/IIIChronic ischemic heart failurePolandCIRCULATEClinical trial phase II/IIIAcute myocardial infarctionPolandABC THERAPYClinical trial phase I/IICutis laxi/ScarsPolandABC THERAPYMedical experimentDiabetic footPolandCD19_CAR-TClinical trialCancerPolandDystrogenMedical experimentDuchenne dystrophy Abbreviations: ALS, amyotrophic lateral sclerosis; ARDS, acute respiratory distress syndrome; COVID-19, coronavirus disease 2019; GvHD, graft versus host disease; HIE, hypoxic-ischemic encephalopathy; MSC, mesenchymal stem/stromal cells; PoC, proof of concept.MethodsSelected new developments are presented in detail: BIOOPA for treatment of epidermolysis bullosa and CartiCure for treatment of cartilage damage in knee joint.ResultsThe BIOOPA dressing for the treatment of epidermolysis bullosa is safe and effective, as no infections or necrosis were observed at the graft implant site during the follow-up period. These patients also reported reduced pain and improved quality of life. When used in orthopedics (CartiCure for cartilage damage), cord tissue-derived MSCs have been shown to be safe and fully integrated into the patient's cartilage, resulting in significant improvement in pain scores and quality of life.DiscussionThe trials presented demonstrate that allogeneic cell therapeutics are the future of regenerative medicine. Use of allogenic technology is the only way to overcome the limitations of the autologous approach: manufacturing process extensively scalable, drug available "off the shelf", applicable in all medical facilities, and economical manufacturing process due to unlimited scalability. Large pharmaceutical companies are investing significantly in this space.

Abstract 30 Should Cord Blood Unit Distribution Patterns Impact Collection Strategies?

IntroductionThe National Cord Blood Program has a large inventory of clinical cord blood units (CBUs) available for transplant (more than 60,000). As a small proportion is distributed each year, new collections should theoretically replenish the inventory, but it is unclear which characteristics should be prioritized for banking.ObjectiveOur goal was to characterize distributed CBUs (in terms of cellularity, race/ethnicity, and match), compare them with the inventory, and devise a collection strategy.MethodsShipments from January 2015 to March 2022 were reviewed (n = 1,046). Race/ethnicity information on CBUs was obtained at the time of collection and was available for 55,934 stored CBUs and 1,020 distributed CBUs. It was also obtained for 394 recipients. HLA match was determined as HLA-A and B antigens and DRB1 allele match (/6), and allelic match as HLA-A, B, C, DRB1 allele match (/8). Total nucleated cells (TNC) and CD34 cells were counted at banking of the CBU, and cell doses (per kg of recipient’s weight) were calculated at shipment.ResultsThe proportion of White Non-Hispanic (WNH) CBUs distributed (38%) was lower than what is present in the inventory (46%), but match levels were higher among these units (Figure 1A and 1B), with 55% of WNH CBUs being 5/8 or higher, vs 40% for minorities’ CBUs. Race match between CBU and recipient led to higher matched transplants, but 41% of race mismatched CBUs still reached 6/8 or higher allele match. The median TNC was 102 × 107 for stored CBUs and 167 × 107 for distributed CBUs (Figure 1C). Better matched units have been selected with lower cell contents (Figure 1D).Figure 1.Distribution characteristics.DiscussionCBUs broaden transplant options across ethnic groups, and much focus has been placed on collecting minorities’ CBUs. However, our analysis showed that WNH CBUs are still the largest distributed population, with higher allele match to the recipient (predicting better transplant outcomes), so these CBUs should not be excluded from collection strategies. From the data presented here, the goal for new collections should be to obtain CBUs with higher TNC and CD34 (spreading across all races and ethnicities). These would replenish the pool of high cellularity CBUs that are more often selected for transplant.

Higher Cryopreserved CD34+ Cell Dose Is Associated with Decreased Hepatic Veno-Occlusive Disease/Sinusoidal Obstruction Syndrome after Single-Unit Cord Blood Transplantation in Adults Given Prophylactic Ursodeoxycholic Acid and Intravenous Heparin

Hepatic veno-occlusive disease (VOD), also known as sinusoidal obstruction syndrome (SOS), is one of the most serious complications to occur early after allogeneic hematopoietic cell transplantation (HCT). However, detailed data on VOD/SOS after cord blood transplantation (CBT) are not available. The objective of this retrospective study was to evaluate the incidence, risk factors, and clinical impact of VOD/SOS after single-unit unrelated CBT for adult patients at our institution. We retrospectively evaluated the incidence, risk factors, and outcomes of VOD/SOS after 390 single-unit unrelated CBTs performed in 332 adults under a prophylactic strategy of ursodeoxycholic acid (UDCA) and i.v. heparin at our institution between 1998 and 2021. VOD/SOS was observed in 24 of the 390 CBTs. The cumulative incidence of VOD/SOS was 5.9% at 30 days and 6.2% at 100 days after CBT. Multivariate analysis showed that cryopreserved CD34+ cell dose ≥1.0×105/kg was significantly associated with a decreased risk of VOD/SOS after CBT (hazard ratio [HR], 0.33; 95% confidence interval [CI], 0.12 to 0.91; P=.032). In multivariate analysis, the development of VOD/SOS was significantly associated with higher overall mortality (HR, 6.19; 95% CI, 3.61 to 10.65; P < .001), treatment failure (HR, 4.79; 95% CI, 2.95 to 7.76; P < .001), and nonrelapse mortality (HR, 12.60; 95% CI, 6.90 to 23.00; P < .001). Our study shows that the incidence of hepatic VOD/SOS was relatively low after unrelated single-unit CBT under a prophylactic strategy of UDCA and i.v. heparin. A higher cryopreserved cord blood CD34+ cell dose was associated with a reduction in VOD/SOS, suggesting that selection of a higher cord blood unit CD34+ cell dose could be efficient in preventing hepatic VOD/SOS in adults undergoing single-unit CBT.

Public Cord Blood Banks as a source of starting material for clinical grade HLA-homozygous induced pluripotent stem cells

BackgroundThe increasing number of clinical trials for induced pluripotent stem cell (iPSC)-derived cell therapy products makes the production on clinical grade iPSC more and more relevant and necessary. Cord blood banks are an ideal source of young, HLA-typed and virus screened starting material to produce HLA-homozygous iPSC lines for wide immune-compatibility allogenic cell therapy approaches. The production of such clinical grade iPSC lines (haplolines) involves particular attention to all steps since donor informed consent, cell procurement and a GMP-compliant cell isolation process.MethodsHomozygous cord blood units were identified and quality verified before recontacting donors for informed consent. CD34+ cells were purified from the mononuclear fraction isolated in a cell processor, by magnetic microbeads labelling and separation columns.ResultsWe obtained a median recovery of 20.0% of the collected pre-freezing CD34+, with a final product median viability of 99.1% and median purity of 83.5% of the post-thawed purified CD34+ population.ConclusionsHere we describe our own experience, from unit selection and donor reconsenting, in generating a CD34+ cell product as a starting material to produce HLA-homozygous iPSC following a cost-effective and clinical grade-compliant procedure. These CD34+ cells are the basis for the Spanish bank of haplolines envisioned to serve as a source of cell products for clinical research and therapy.

Frequency and distribution of HLA-DQB1 alleles from 2076 cord blood samples of the Vietnamese cohort.

Human leucocyte antigen (HLA) alleles are very diverse and characterized by ethnicity. To date, information about the frequencies and distributions of HLA alleles among the Vietnamese population is still limited. In this study, HLA-DQB1 alleles of 2076 cord blood units from individuals belonging to Vietnam's Kinh ethnic people were genotyped using Luminex-based polymerase chain reaction sequence-specific oligonucleotide. The results of the study demonstrated that there were 23 alleles on the locus HLA-DQB1. Among those, there were six alleles with high frequencies of over 5%, including DQB1* 03:01 (35.9%), DQB1* 05:01 (12.8%), DQB1* 03:03 (12.2%); DQB1* 06:01 (7.20%), DQB1* 05:02 (6.62%) and DQB1* 02:01 (5.30%) and five rare alleles with low frequencies of below 0.1%. More importantly, this study for the first time reported the presence of two new rare alleles including DQB1* 01:01 and DQB1* 01:02. Conclusively, this study provided significant information about HLA-DQB1 alleles for further investigations and clinical applications.

Favorable Outcome with Conditioning Regimen of Flu/Bu4/Mel in Acute Myeloid Leukemia Patients in Remission Undergoing Cord Blood Transplantation

Cord blood transplantation (CBT) is a curative therapeutic option for patients with acute myeloid leukemia (AML) who do not have an HLA-matched donor. The decline in early nonrelapse mortality (NRM) after CBT has significantly improved overall survival (OS) during the past 20 years because of advances in CBT practices, including more careful patient selection, use of safer conditioning regimens, better cord blood unit selection, and improved supportive care. A previous study reported a conditioning regimen comprising fludarabine, busulfan, and melphalan (Flu/Bu4/Mel) developed for patients undergoing CBT in non-complete remission (CR) myeloid malignancies that showed durable engraftment and remission with acceptable nonrelapse mortality (NRM), leading to excellent survival outcomes. However, no prior study has focused on the role of Flu/Bu4/Mel in CBT conditioning and compared it with conventional myeloablative conditioning (MAC) for AML patients in CR. We aimed to investigate the efficacy and safety of Flu/Bu4/Mel compared with cyclophosphamide and total body irradiation (CY/TBI)-based MAC for AML patients in CR who underwent CBT. Patients were selected from the Japanese nationwide transplantation registry according to the following inclusion criteria: (1) patients with AML aged ≥16 years, (2) first single-unit CBT, and (3) CR at the time of transplantation. Of 477 eligible patients, 148 (31.0%) received CY/TBI, 223 (46.8%) received high-dose cytarabine (HDCA)/CY/TBI, and 106 (22.2%) received Flu/Bu4/Mel. The probability of OS at 3 years was 64.8% (95% confidence interval [CI], 56.0% to 72.3%) in the CY/TBI group, 65.1% (95% CI, 57.8% to 71.4%) in the HDCA/CY/TBI group, and 65.5% (95% CI, 53.7% to 74.9%) in the Flu/Bu4/Mel group (P=.71); the cumulative incidence of relapse at 3 years was 22.0% (95% CI, 15.2% to 29.5%), 17.2% (95% CI, 12.2% to 22.9%), and 18.0% (95% CI, 11.2% to 26.2%), respectively (P=.40); and the cumulative incidence of NRM at 3 years was 17.2% (95% CI, 11.5% to 24.0%), 20.7% (95% CI, 15.4% to 26.7%), and 18.6% (95% CI, 11.4% to 27.2%), respectively (P=.95). Multivariate analysis identified Flu/Bu4/Mel as a favorable factor for OS; however, it was not significantly favorable for relapse and NRM in the CY/TBI, HDCA/CY/TBI, and Flu/Bu4/Mel groups (hazard ratio [HR], .50 [95% CI, .29-.88], P=.015; .67 [95% CI, .31-1.46], P=.31; and .55 [95% CI, .26-1.18], respectively; P=.12). Flu/Bu4/Mel was a favorable factor for neutrophil engraftment (HR, 1.51; 95% CI, 1.08 to 2.12; P=.016). Multivariate analysis showed that Flu/Bu4/Mel had a favorable prognostic impact on OS and neutrophil engraftment despite the non-TBI regimen. Our findings suggest that Flu/Bu4/Mel may sustain the antileukemia effect with decreasing NRM and could be a favorable CBT conditioning regimen for patients with AML in CR.

A comprehensive human leukocyte antigen analysis of 36 782 cord blood units stored in the Australian Public Cord Blood Banking Network

Background and aimsThe network of public cord blood banks (CBBs) in Australia, known as AusCord, comprises CBBs located in Brisbane, Sydney and Melbourne. A novel comprehensive analysis has been performed to determine whether the cryopreserved, searchable cord blood unit (CBU) inventory of approximately 36 000 units share similar tissue types or haplotypes. MethodsHuman leukocyte antigen (HLA) data was analysed using Microsoft Excel following standardisation of typing data. ResultsThe analysis has found that the majority of stored, searched and released CBU exhibit a tissue type that is unique within and between the CBBs. Therefore, each collection performed by the CBBs is likely to comprise a tissue type that is not already stored among the total AusCord inventory. HLA alleles (HLA-A*34, HLA-B*56, HLA-DRB1*08:03), which are uncommon in European populations, were associated with Pacific Islander and/or Indigenous Australian populations and confirmed to be more frequent among donors who, when screened, self-identified as these ethnicities. ConclusionsThese data indicate that (i) continued addition of CBU to existing inventories is likely to further increase the HLA diversity and (ii) screening donors for ethnicity or strategically locating collection sites where ethnic minorities reside can successfully result in collection of rare HLA associated with ethnic minority groups for whom finding donors might otherwise be more difficult.