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Background: HCT is the treatment of choice for many children with non-malignant diseases. The aim of this study is to determine major transplant outcomes among pediatric patients (pts) transplanted in Brazil. Patients and methods: This retrospective study included 300 pts with Bone Marrow Failures syndromes (65%, n = 193), Primary Immunodeficiencies (24%, n = 72), Inborn Errors of Metabolism (7%, n = 22) and Hemoglobinopathies (4%, n = 13) who received first allo-HCT between 2010 and 2014 at 11 Brazilian BMT centers. All pts received unmanipulated bone marrow (BM, 86%) or cord blood (CB, 14%) grafts. HCT donors included match related (MRD, n = 140, 47%), fully matched unrelated (MUD, n = 108, 36%) and mismatched unrelated (MM-URD, n = 52, 17%). Reduced intensity conditioning was given to 167 pts (56%). Most (78%) received GVHD prophylaxis with calcineurin inhibitor plus methotrexate. Grading of aGVHD and cGVHD were per Glucksberg and NIH criteria and were uniform among all centers. Results: Median age was 8 y/o (range, .6 to 17) and 62% (n = 185) were male. With median follow up of 4.5 years, 4yoverall survival (OS) was better for MRD compared to matched URD or mismatched URD transplants (84% versus 71% versus 69%, respectively; P = .01). When compared MM-URD BM (n = 26), matched (n = 11) or mismatched unrelated CB (n = 26) transplant pts had a lower OS (HR = 2.73; P = .04). Twentytwo pts died before D+21 and therefore were not evaluable for engraftment. Primary graft failure (GF) was observed in 21 pts and was significantly higher after mismatched CB transplants (OR = 7.43; P = .03) compared to MM-URD BM transplants. 257 pts engrafted and 22 developed secondary GF at a median of 193 days (range, 39-431). Acute GVHD grade II-IV occurred in 47 pts at a median of 28 days after HCT with a cumulative incidence (CumInc) at 3 months of 16% (95% CI, 11-20). Chronic GVHD was reported in 60 pts at a median of 189 days after HCT with a CumInc at 2 years of 20% (95% CI, 15-24). Acute and chronic GVHD CumInc were not significantly different across all donor types. Seventy-three pts died at a median of 69 days after HCT. Major causes of death included infection (51%) and GVHD (15%). One-year and 100days TRM were 20% (95% CI, 15-24) and 17% (95% CI, 13-22), respectively. Conclusions: Children and adolescents with non-malignant diseases can achieve excellent outcomes after HCT from MRD in Brazil. We observed a high early mortality after HCT from unrelated CB donors in our study. For children lacking MRD, an URD donor is a suitable option, particularly from a MUD. Confirming earlier reports, incidence of chronic GVHD was low in this setting. This is the first pediatric multicenter study in Brazil for nonmalignant diseases and future studies will focus on strategies to reduce early mortality after unrelated CB HCT and will explore the role of HCT from haploidentical donors for children lacking MRD or MUD.

Background: As part of routine quality monitoring of frozen hematopoietic progenitor cell (HPC) products we perform Trypan Blue (TB) viability upon thawing. The sixth edition of FACT standards requires an assay for viable CD34 be performed on HPC products intended for hematopoietic reconstitution (D8.1.3.2). The eighth edition of AABB standards also requires a test for viability (5.17A.4.a). TB dye exclusion is a simple and rapid test for differentiating total living cells from nonviable cells, but dye uptake assessment can be subjective and CD34 specific measurement is not possible. Measuring cell viability by flow cytometry (flow) using the flurochrome 7-amino actinomycin (7-AAD) is less vulnerable to subjectivity and allows for cell-type specific assessment, but is more expensive, time-intensive, and requires specialized laboratory equipment and advanced technical expertise. Therefore, we evaluated TB compared to CD45 7-AAD viability, and CD45 7-AAD compared to CD34 7-AAD viability in order to establish that TB is an acceptable alternative to CD34 viability for frozen-thawed HPC collected by apheresis HPC(A). Methods: TB viability was performed with TB solution (.4%) with HPC(A) sample re-suspended in alpha Minimum Essential Media. Samples submitted to the flow lab were aliquoted from HPC(A) products suspended in dimethyl sulfoxide, then washed twice in buffer and stained within one hour of receipt by the flow lab. Analysis used the International Society of Hematotherapy and Graft Engineering (ISHAGE) protocol. The mean viability, mean difference, 95% confidence interval (CI) were calculated, and statistical significance was determined using the paired t-test. Results: For frozen-thawed HPC(A) samples (n = 39), mean CD45 7-AAD viability was 82.4% (CI 79.9 to 84.9%), and mean TB viability was 80.0% (CI 78.0 to 82.0%). The average difference is 2.4% which is not statistically significant (P = .12). Mean CD45 7-AAD viability was 82.3% (CI 79.6 to 84.9%) and mean CD34 7-AAD viability was 96.9% (CI 95.8 to 98.0%) (n = 33) where CD34 7-AAD viability was consistently greater than CD45 7-AAD viability by 14.6% (mean, CI 12.1 to 17.1%). Conclusion: For frozen-thawed HPC(A), overall viability measured with TB is not significantly different from viability measured by CD45 7-AAD. Furthermore, CD34 7-AAD viability is always greater than CD45 7-AAD viability. Therefore, we established that our process using TB viability for quality assessment of frozen-thawed HPC(A) products is an acceptable alternative to CD34 viability of each product and in compliance with D8.1.3.2 and 5.17A.4.a.

Background:The initial HCT evaluation can be overwhelming for a patient and caregiver. The appointment involves visits with the multi-disciplinary team who communicate an extensive amount of information. The patient and caregiver are given valuable written material of the HCT process that reinforces the verbal discussion. Even with written and verbal communication tools, patients reported gaps in knowledge about the HCT process. Patients also report feeling overwhelmed with the amount of verbal and written material presented. In addition, education teaching and method dispersed to patients and their caregivers vary among HCT Nurse Coordinators. Objectives: The aims of the project were to: 1) Increase the patient's and caregiver's understanding of HCT through a visual and oral presentation tool; 2) Increase patient's and caregiver's retention and knowledge level of HCT process; and 3) Ensure standardization of the patient education material taught by the HCT Nurse Coordinator. Method: A power point presentation was created for use during the first HCT evaluation visit. The HCT Nurse Coordinator accessed the presentation and referred to the images and photographs during the discussion of HCT. Included in the presentation, but not limited to, are images of stem cells, central lines, the hemapheresis department and equipment, and the injection teaching process. Pictures of our outpatient and new 23-bed inpatient HCT units were also included. Findings: Visualizations help to increase patient knowledge about line placement, hemapheresis, and the HCT process. Also, providing photographs of critical HCT processes for retention of material decreases anxiety and allows patients to be better prepared for HCT. By utilizing the power point presentation during the initial evaluation visit, the HCT Nurse Coordinators can consistently provide the same information taught to each patient. Conclusion: The addition of a visual presentation tool during the initial HCT evaluation visit can reinforce verbal and written resources for patients and caregivers. By utilizing the visual presentation tool, HCT Nurse Transplant Coordinators can provide consistent information regarding the HCT process and increases the patient's knowledge level.

HCT; 162 with a MUD. For those who identified a MUD and proceeded to HCT, median time between FS and date of confirmatory donor HLA testing was 16 days (range 3-540). Median time from FS to donor information session was 58 days (range 1-645), FS to start of preparatory regimen (PR), 92 days (range 8-772), and FS to infusion, 107 days (range 22-786). An alternative donor source was used for 28 patients [22 CBU, 5 matched sibling, 1 haplo] when no MUD was identified. Additionally, 110 patients did not undergo HCT. The reasons for not proceeding with HCT were patient death (27), HCT not indicated (25), alternate therapy chosen (19), no suitable donor (15), family preference (2), and patient deemed ineligible (1). Eleven patients are still undergoing diagnostic evaluation, 4 are undergoing pre-HCT evaluation, and 6 were lost to follow-up. Our data show that under the best circumstances, a MUD can be identified quickly, with 44% of our patients moving from FS to confirmatory donor testing within 14 days. Time to PR and infusion is often influenced by variability in patients' condition. In cases where a donor is needed urgently but the search is not ideal, we have used haplo HCT. MUD could not be identified in only 5% of our last 300 searches and haplo HCT is of value in these patients.