Quality Assessment of Cryopreserved Peripheral Blood Stem Cell Products: Evaluation of Two Methods for Flow Cytometric Viability Testing.

Introduction: The standard flow cytometry method for viability testing using 7-aminoactinomycin D (7-AAD) determines cells in necrosis and late apoptosis. The colony-forming unit (CFU) assay, which evaluates the proliferation ability of HSCs, is also used in graft quality assessment despite known deficiencies that make this assay impractical in routine clinical settings. The aim was to compare the effectiveness of the flow cytometry 7-AAD/annexin V method with the 7-AAD method in assessing the quality of HSCs in autologous and allogeneic peripheral blood stem cell (PBSC) products.

Influence of donor characteristics on the milk donation volume and microbiological quality of donated human milk in first 3 years of work of the Croatian human milk bank-retrospective observational study.

Objectives: Donated human milk is the best possible alternative when mother's own milk is not available. The aim of this study is to investigate whether there are differences in the milk donation volumes and microbiological quality of donated milk depending on human milk donors (HMDs) characteristics.

Methods: We analyzed data on the HMDs who donated milk in the first three years of work of human milk bank (HMB) - November 2019 to January 2023.

The impact of cryoprotectant exposure time on post-thaw viability of autologous and allogeneic hematopoietic stem cells and leukocyte subpopulations.

Although the use of cryoprotectant dimethyl sulfoxide (DMSO) is the gold standard in cryopreservation of hematopoietic stem cells, it is well known that it has a negative effect on cell viability. The aim of this prospective study was to examine how the length of post-thaw exposure to DMSO affects the cell viability and stability of peripheral blood stem cell (PBSC) samples. Additionally, the effects of donor type and pre-cryopreservation storage time on post-thaw viability during the stability study were evaluated.

Vascular access for autologous peripheral blood stem cells collection by large volume leukapheresis: Single center experience.

Introduction: Peripheral blood stem cell (PBSC) harvesting requires reliable and safe vascular access. In our institution, a change of practice was implemented and the central venous catheter (CVC) placement for all autologous PBSC collections was abandoned in favor of a careful evaluation of peripheral venous access (PVA) for each individual patient. The aim of this prospective study was to evaluate the rate of patients with adequate peripheral veins for autologous PBSC collection and compare patient characteristics, collection efficacy, and complication rate between patients with PVA and CVC.

Variable recovery of cryopreserved hematopoietic stem cells and leukocyte subpopulations in leukapheresis products.

Introduction: Due to the expansion of cell therapy using not only haematopoietic stem cells (HSC) but also other leukocyte subpopulations, the loss of these cells in cryopreserved apheresis products needs to be evaluated. Various factors that could negatively affect post-thaw recovery, such as leukapheresis product characteristics, storage time and cryopreservation protocols have been identified.

Methods: The post-thaw recovery of HSCs, lymphocytes, NK cells and monocytes, as well as the factors that could adversely affect it were analysed in autologous and allogeneic leukapheresis products.

36 Cornea microbiology contamination rate depending on post mortem time, retrospective analysis at croatian tissue and cell bank, UHC Zagreb.

Background: Corneas procured post mortem are at risk of microbiology contamination, therefore decontamination procedures before storage, aseptic techniques during processing and antimicrobials used in the storage medium are routinely used. Despite that, corneas are discarded due to microbiology contamination. According to professional guidelines, corneas can be procured preferably within 24 hours after cardiac arrest but up to 48 hours.

The appropriateness of platelet transfusions in hematological patients and the potential for improvement.

Background: Hematology patients are intensive platelet users. In clinical practice, a substantial proportion of platelet (PLT) transfusions are routinely administered outside the guidelines despite compelling evidence for recommendations. Those unnecessary PLT transfusions are an unjustified extra burden on a scarce healthcare resource and may also be detrimental to the patients.

Cryopreserved human heart valve allografts: a ten-year single centre experience.

This study provides an overview of tissue banking activities at the Croatian Cardiovascular Tissue Bank (CTB) during past ten years and presents the outcomes of cryopreserved heart valve allografts (CHAs) use in different patient groups. From June 2011 until December 2021, 75 heart donations were referred to CTB: 41 recipient of heart transplant (RHT), 32 donors after brain death (DBD) and 2 donors after circulatory death (DCD) donations. Processing resulted in 103 valves of which 65 met quality requirements for clinical use.

Severe intracranial haemorrhage in neonatal alloimmune thrombocytopenia due to antibodies against human platelet antigen 1b: Case report and literature review.

Fetal/neonatal alloimmune thrombocytopenia (FNAIT) is a rare life-threatening disorder, leading to severe thrombocytopenia and potentially bleeding, with intracranial haemorrhage (ICH) being the most serious complication. We report on a FNAIT case with fourth-degree ICH that arose due to antibodies against human platelet antigen (HPA)-1b. The male infant, born to an otherwise healthy mother, presented with severe signs of ICH soon after delivery.

Changes in the incidence of transfusion reactions in hematological patients over the past 30 years.

Background: Patients with hematological diseases are polytransfused and often immunocompromised, therefore susceptible to transfusion reactions (TR). This study aims to document the incidence of TRs in adult hematological patients and assess the effect of changes in the production of blood components and transfusion practice on their occurrence.

Study Design And Methods: Retrospective observational analysis of TRs reported from 1993 to 2019 was performed.

The role of serological and molecular testing in the diagnostics and transfusion treatment of autoimmune haemolytic anaemia.

Autoimmune haemolytic anaemia (AIHA) is a rare autoimmune disease characterised by haemolysis associated with the presence of immunoglobulins and/or components of the complement system on red blood cells (RBCs). It is classified into warm or cold antibody-mediated AIHA according to the temperature at which autoantibodies bind optimally to RBCs. Clinicians should be familiar with the procedural tests used for a complete laboratory investigation of AIHA.

Bleeding risk stratification in coronary artery surgery: the should-not-bleed score.

Background: An estimated 20% of allogeneic blood transfusions in the United States are associated with cardiac surgery. It is estimated that 11% of red cell resources were used for transfusion support of patients undergoing coronary artery bypass grafting (CABG) with a documented wide variability in transfusion rate (7.8 to 92.

Evaluation of the BD Stem Cell Enumeration Kit on the BD FACSCanto II flow cytometer using bd facscanto clinical and bd facsdiva software.

Introduction: CD34+ hematopoietic stem cell (HSC) enumeration by cell flow cytometry is routinely used in clinical laboratories for monitoring of HSC mobilization into peripheral blood and assessment of the quality of HSC products. The modified ISHAGE protocol is the most often used procedure for determination of CD34+ cells using flow cytometry. The aim of this study was to evaluate BD Enumeration stem cell kit on flow cytometer BD facscanto II, using facscanto clinical and facsdiva softwares.

K antigens on neonatal red blood cells blocked by anti-K with titer of 32.

The blocking of red blood cell (RBC) antigens occurs when potent maternal antibodies bind to antigens on fetal or neonatal RBCs, causing them to be negative when typed with human IgM antisera. This phenomenon is rare; when it does occur, the antibody is usually of a high titer. This reported finding is typically due to anti-D, with rare reports describing false-negative K phenotyping due to blocking by maternal anti-K.

The Outcome of Hemolytic Disease of the Fetus and Newborn Caused by Anti-Rh17 Antibody: Analysis of Three Cases and Review of the Literature.

Background: Anti-Rh17 is a rare red blood cell (RBC) antibody to high-frequency antigens that may cause severe hemolytic disease of the fetus and newborn (HDFN). Despite the rarity of HDFN caused by Anti-Rh17, this antibody was reported in many different populations. Emergency transfusions, especially exchange transfusions, present a huge problem if no compatible RBCs of phenotype D- are available.

LEUKOCYTAPHERESIS IN THE MANAGEMENT OF SEVERE STEROID-DEPENDENT ULCERATIVE COLITIS.

Ulcerative colitis (UC) is a multifactorial disease of unknown precise etiology and immunopathogenesis. Peripheral blood granulocytes and monocytes/macrophages are the major sources of cytokines, which regulate inflammation. Leukocytapheresis (LCAP) is a method where blood is processed by apheresis system that removes lymphocytes and plasma before being returned to the body.

Recurrent Aortic Infections Due to Unrecognized Aorto-Enteric Fistula.

We report a case of an infrarenal abdominal aortic aneurysm (AAA) with unrecognized primary aortoduodenal fistula (ADF), treated by endovascular aortic repair (EVAR). Endograft infection was diagnosed 12 months thereafter. The associated ADF was uncovered during open surgery, which included endograft extraction, in situ aortic reconstruction with a cryopreserved homograft (CHG) and duodenal repair.

Cost Analysis of Transfusion Therapy in Coronary Artery Surgery.

Background:  In patients undergoing coronary artery bypass grafting (CABG), wide variability in transfusion rate (7.8% to 92.8%) raises the question of the amount of unnecessary transfusions.

Performance prediction algorithm for autologous PBSC collection in adults and pediatric patients using large volume leukapheresis.

Background And Objectives: The number of CD34+ cells collected in apheresis procedures depends mainly on the collection efficiency of the device and the blood volume processed. Large volume leukapheresis (LVL) can improve CD34+ cell yield and has previously been investigated using the COBE Spectra device (Terumo BCT, USA).

Materials And Methods: This was a retrospective analysis of LVL performance in patients undergoing continuous mononuclear cell collection (CMNC) using the new Spectra Optia apheresis system (Terumo BCT, USA) at the University Hospital Center, Zagreb, from March 2016 to September 2016.

An automated mini buffy coat preparation method for use in mini extracorporeal photopheresis treatment of graft-vs-host-disease in a low body weight pediatric patient.

A mini extracorporeal photopheresis (mini-ECP) "off line" technique has been developed for use in the treatment of small children and patients with apheresis contraindications. Until now various methods have been used for buffy coat separation from whole blood. In this report we describe a protocol for mini buffy coat preparation using the automated Sepax laboratory separator for "off line" ECP treatment in a low body weight child with graft-vs-host-disease.

Susceptible and protective HLA-DR and HLA-DQ alleles for Fy alloimmunization in the Croatian population.

Background: Alloimmunization is a known risk of transfusion therapy caused by exposure to foreign RBC antigens. However, alloimmunization is not observed in all transfused patients. Human leukocyte antigen (HLA) molecules may contribute to the recognition and presentation of foreign antigens and to the potency of immune responses that result in the production of antibodies.

Comparative Analysis of Biological and Functional Properties of Bone Marrow Mesenchymal Stromal Cells Expanded in Media with Different Platelet Lysate Content.

Due to their ability to induce immunological tolerance in the recipient, mesenchymal stromal cells (MSCs) have been utilized in the treatment of various hematological and immune- and inflammation-mediated diseases. The clinical application of MSCs implies prior in vitro expansion that usually includes the use of fetal bovine serum (FBS). The present study evaluated the effect of different platelet lysate (PL) media content on the biological properties of MSCs.