Long-Term Cryopreservation of Peripheral Blood Stem Cell Harvest Using Low Concentration (4.35%) Dimethyl Sulfoxide with Methyl Cellulose and Uncontrolled Rate Freezing at -80 °C: An Effective Option in Resource-Limited Settings.

Long-term cryopreservation of peripheral blood stem cells (PBSCs) is highly useful in the setting of tandem/multiple transplantations or treatment of relapse in the autologous hematopoietic stem cell transplantation (HSCT) setting. Even in allogeneic HSCT, donor lymphocyte infusions may be stored for months to years if excess stem cells are collected from donors. Cryopreservation is a delicate, complex, and costly procedure, and higher concentrations of dimethyl sulfoxide (DMSO), a commonly used cryoprotectant, can be toxic to cells and cause adverse effects in the recipient during infusions. In this study, we examined the effect of long-term cryopreservation using 4.35% DMSO (as final concentration) with methyl cellulose and uncontrolled rate freezing in a mechanical freezer (-80 °C) on the viability and colony-forming ability of CD34+ human PBSCs. For patients undergoing autologous HSCT, PBSCs were cryopreserved using DMSO (final concentration of 4.35%) with methyl cellulose. The post-thaw viability of PBSCs was determined using Trypan blue exclusion and flow cytometry-based 7-amino-actinomycin-D (FC-7AAD) methods. Concentrations of CD34+ stem cells and immune cell subsets in post-thaw PBSC harvest samples were assessed using multicolor flow cytometry, and the clonogenic potential of post-thaw stem cells was studied using a colony-forming unit (CFU) assay. CD34+ stem cell levels were correlated with the prestorage CD34 levels using the Pearson correlation test. The viability results in the Trypan blue dye exclusion method and the flow cytometry-based method were compared using Bland-Altman plots. We studied 26 PBSC harvest samples with a median cryopreservation duration of 6.6 years (range, 3.8 to 11.5 years). The median viability of post-thaw PBSCs was >80% using both methods, with a weak agreement between them (r = .03; P = .5). The median CD34+ stem cell count in the post-thaw samples was 9.13 × 106/kg (range, .44 to 26.27 × 106/kg). The CFU assay yielded a good proliferation and differentiation potential in post-thaw PBSCs, with a weak correlation between granulocyte macrophage CFU and CD34+ stem cell levels (r = .4; P = .05). Two samples that had been cryopreserved for >8 years showed low viability. Cryopreservation of PBSCs using 4.35% DMSO with methyl cellulose and uncontrolled freezing in a mechanical freezer at -80 °C allows the maintenance of long-term viability of PBSC for up to 8 years.

The determinants of donor safety and product quality in optimization of apheresis granulocyte harvest: An experience from a tertiary care oncology centre in India.

Allogeneic granulocyte transfusions play a substantial role in treatment of lifethreatening neutropenia-associated infections in patients undergoing intensive chemotherapy and hematopoietic stem cell transplant. Granulocyte harvest by apheresis is considered a safe and effective method to obtain adequate therapeutic granulocyte dosage for clinical effectiveness. This study described the experiences associated with apheresis granulocyte harvest procedures in our tertiary care haemato-oncology centre. We have analysed the incidence of adverse events (AEs) with associated potential risk factors contributing to donor safety and improvement in product quality. Retrospective data of 131 healthy allogeneic donors who underwent granulocyte harvest from May 2016 to July 2020 were analyzed. AEs were observed in overall 29 procedures (22.13%), including 14.50% citrate reactions, 7.6% venous access-related reactions, and 1.52% vasovagal reactions. Older age (p = 0.012) and higher body mass index (p = 0.015) in donors were significant variables found associated with a higher incidence of AEs. There was no significant impact of AEs on granulocyte product yield (p = 0.41) with a median collection yield of 1.73 × 10 10 cells/ unit. In multivariate analysis, post-mobilization parameters like total leukocyte counts (p = 0.036), absolute neutrophil counts (p = 0.042), and platelet counts (p = 0.006) showed a positive correlation with higher product yield. All the donors successfully donated and tolerated granulocyte colony stimulating factor plus dexamethasone mobilization and granulocyte apheresis harvest without any serious AEs. Our study shows that optimal technical and procedural modifications during apheresis granulocyte harvest procedures can overcome the associated potential risks by providing donor safety and improving product quality.

Incidence and Analysis of 7 Years Adverse Transfusion Reaction: A Retrospective Analysis.

Safe blood transfusion is the primary need of all the health care delivery system. Though with the advances of transfusion medicine, the incidences of transfusion risk is gradually reduced, but the adverse transfusion reaction (ATR) of non hemolytic type still prevails. The purpose of this study was to estimate the incidence and pattern of transfusion-related adverse events at our centre. The present retrospective observational study was conducted in the Department of Transfusion Medicine from April 2011 to April 2018, at a multi-organ transplant centre in South India. All the Adverse transfusion reactions were investigated in detail in the blood bank for the clerical errors, immunohematology workup and classified according to their nature with imputability assessment. A total of 140 ATR were reported out of 100,569 blood components distributed during the study period. After the analysis and workup of the reported reactions, majority of the reactions were observed in males (71%, n = 99). Most common symptom presented was Itching/Rashes in 43.6% (n = 61) ATR. Allergic reactions (51.4%, n = 72), were the most commonly encountered ATR followed by FNHTR (25.7%, n = 36). FFP transfusions (0.2%) contributed to the majority of the reactions followed by Red cell transfusion (0.15%). ATR were observed maximum in Hepato-biliary disease and liver transplantation patients (62%) followed by oncology patients (15%). The overall incidence of ATR in our study is 0.14% which is comparatively low compared to other studies due to well established hemovigilance systems. Adoption of more equipped methods & sensitive technology in various areas of blood banking will help to bring down the unwanted adverse transfusion reactions.

A rare Bombay (Oh) phenotype to 'A' blood group - Live donor liver transplant.

Bombay (Oh) phenotype is the rarest blood group in India characterized by the absence of A, B, and H antigens and the presence of anti-H antibodies besides anti-A and anti-B. There is no literature predicting the safety of Oh blood group organ donation to non-Oh blood group recipient. We present the first reported case of successful live donor liver transplantation from an Oh-positive liver donor to an A-positive blood group recipient with hepatitis B virus-related chronic liver disease. The case highlights the need for proper immunohematological workup, national registry of rare group blood donors and need of protocol for perioperative monitoring and blood management in ABO-incompatible organ transplants involving Oh group donor or recipient.