Reinhard Henschler

Background and Objectives: Global demand is rising for human platelet lysate (HPL) in the manufacture of cell‐based medicinal products, but platelet resources are scarce. As protocols for the production process and quality testing are heterogeneous, the Working Party on Cellular Therapies of the International Society of Blood Transfusion (ISBT) has recently conducted a survey to obtain information from blood establishments worldwide. Materials and Methods: The survey was created at limesurvey.net and consisted of 29 questions on the use of platelet concentrates (PCs) for HPL production to determine the status, conditions, standardization efforts, area of application, and potential difficulties. The link was sent to all ISBT members, and data were analyzed descriptively. Results: Of 78 participating centers, 12 routinely produce HPL according to in‐house protocols, mainly by pooling expired PCs, whereas quality control testing is established in 10 institutions. HPL is used for basic research ( n = 7) and for the manufacture of cell‐based medicinal products ( n = 7). Conclusion: HPL manufacture is still heterogeneous and limited to a few blood centers worldwide. Prospective collaborations between these institutions may contribute to the efforts for international harmonization and consistent regulation of HPL applications.

Background and Objectives: Hypoxic/hypocapnic (HH) treatment and storage conditions have been shown to reduce oxidative stress and improve red blood cell (RBC) quality. This study aimed to validate a good manufacturing practice HH RBC product for obtaining the licence for routine use in patients, by comparison with normoxic RBCs both without irradiation and after gamma irradiation. Materials and Methods: Forty‐four units of leucoreduced red cell concentrates (LR‐RCCs) were prepared from 500 mL citrate–phosphate–dextrose (CPD)/phosphate‐adenine‐glucose‐guanosine‐saline‐mannitol (PAGGSM) whole‐blood donations. Paired units of ABO‐matched LR‐RCCs were pooled and split into equal aliquots. Twenty‐two units (control) were maintained at room temperature for 3 h, while 22 units underwent HH treatment for 3 h using the Hemanext ONE® system. Eleven control and 11 HH LR‐RCCs were assigned to the non‐irradiated arm, and the remaining 22 units (11 control, 11 HH) were irradiated on Day 14. In vitro RBC quality was measured until Day 42 (non‐irradiated) and Day 28 (irradiated on Day 14). Results: The HH product was comparable to controls in haemoglobin (Hgb) content, haematocrit, potassium and haemolysis, both without irradiation over 42 days and after irradiation over 28 days. HH RBCs contained 1.2‐fold higher adenosine triphosphate (ATP) and up to 6.2‐fold higher 2,3‐diphosphoglycerate (2,3‐DPG) levels. Lactate was 25% increased, and glucose levels were up to 30% decreased compared with normoxic RBCs. Conclusion: HH storage enhanced RBC ATP and 2,3‐DPG levels and yielded a product fulfilling RBC quality criteria for the other parameters. These findings support its clinical use for optimizing RBC transfusion quality, particularly for irradiated units.

Background: Refrigerated storage of red cell concentrate ( RCC ) leads to metabolic, oxidative, and structural changes that impair functionality and viability. These changes can be attenuated by hypoxic storage. Objective: This study assessed the quality of leucocytes‐reduced ( LR ), O ₂ / CO ₂ reduced red blood cells ( RBC ) stored for 42 days after pre‐storage O ₂ / CO ₂ reduction with the Hemanext ONE System, to verify compliance with regional and blood center acceptance criteria across six sites. Study design and methods: Validation studies of the in vitro performance of the LR , O ₂ / CO ₂ reduced storage (Hemanext ONE ) System were planned and executed at blood production centers in Italy, Germany, Norway and Switzerland. The endpoint‐associated study acceptance criteria included total hematocrit >50%, total hemoglobin ≥40 g and hemolysis <0.8%. Results: In total, 133 whole blood units donated at six blood centers were evaluated after processing with the Hemanext ONE System and after 21 and 42 days of storage. At Day 1 post‐processing, total hemoglobin (mean ± SD ) was 52.4 g/unit ± 3.1 and total hematocrit percentage (mean ± SD ) was 60.6% ± 7.0 (range, 51.9%–81.0%). Hemolysis at Day 42 of storage: overall mean ± SD , 0.26% ± 0.14. Discussion: Hypoxic RBCs met regulatory quality criteria regardless of collection and processing modalities. These results indicate that processing of RBCs stored under hypoxic conditions satisfies acceptance criteria for transfusion into patients in six European regional blood centers.

Background: Apheresis procedure of autologous lymphocytes competent for proliferation and expansion is a crucial step in the production of chimeric antigen receptor (CAR) T‐cells. Previous therapies or disease status prior to collection may negatively impact the collections.Study Design and Methods: We performed a retrospective analysis with the aim to determine cellular factors in association with the collection of autologous T‐cells and subsequent CAR T manufacturing toward tisagenlecleucel (tisa‐cel). Between February 2019 and February 2022, 63 collections of 54 patients were performed for subsequent therapy with tisa‐cel.Results: We observed no difference in median CD3+ cell yields according to the number of prior therapy lines (>3 vs. ≤3, p = .335), prior treatment with bendamustine (p = .954) or marrow infiltration (p = .634). Fifty‐six collections were sent for manufacturing, of which 22 (39%) resulted in manufacturing failures, namely terminations (n = 12) or out‐of‐specification events (n = 10).Collections resulting in manufacturing failures yielded significantly lower CD3+ (p = .005), CD3+CD4+ (p = .044), and non‐senescent CD3+CD27+CD28+ (p = .003) counts. Multivariable analysis identified the absolute number of CD3+CD27+CD28+ cells as relevant, with a calculated cut‐off of ≥34.58 × 10⁸ CD3+CD27+CD28+ cells for 89.5% probability of successful CAR T‐cell production.Discussion: In summary, we report a positive influence of a higher number of non‐senescent Τ‐cells on successful manufacturing. Further analyses are required to determine measures for further optimization of collection outcomes.

Background and Objectives: We should control free K ⁺ during massive transfusion (>80 mL/kg) of red blood cells (RBCs) in small children. To manage this, several national and international guidelines recommend using RBCs stored only up to 7 days. We tested a washing step for RBCs in saline‐adenine‐glucose‐mannitol (SAGM) with or without irradiation to reduce supernatant K ⁺ levels, improve quality and potentially extend the shelf life of stored RBCs. Materials and Methods: RBCs of 240–330 mL were prepared from whole blood donations, then split into halves and stored in SAGM solution at 4 ± 2°C for 21 days. RBCs were split and washed on Days 1 and 8, and some were gamma‐irradiated on Day 8. Glucose, lactate, lactate dehydrogenase (LDH), adenosine triphosphate (ATP), K ⁺ and haemolysis were determined over 21 days. Results: After washing on Day 1, only glucose and lactate improved, whereas after washing on Day 8, LDH and K ⁺ also improved. Irradiation resulted in accelerated K ⁺ accumulation and increased haemolysis. Mean extracellular K ⁺ concentrations were 21.2 ± 1.03 mM after irradiation on Day 8 versus 1.12 ± 0.05 mM after irradiation plus wash on Day 8, and 38.80 ± 2.13 mM on Day 10 after irradiation on Day 8 and 16.6 ± 0.05 mM on Day 10 after irradiation plus wash on Day 8. Conclusion: K ⁺ concentrations remained <25 mM within 8 days of storage. We recommend irradiation by Day 8 at the latest for neonatal transfusion. The shelf life may be extended by another 48 h if the RBCs are also washed.

Intravenously transplanted mesenchymal stromal cells (MSCs) have been shown to interact with endothelial cells and to migrate to tissues. However, intracellular signals regulating MSC migration are still incompletely understood. Here, we analyzed the role of Rap1 GTPase in the migration of human bone marrow-derived MSCs in vitro and in short-term homing in mice in vivo. MSCs expressed both Rap1A and Rap1B mRNAs, which were downregulated after treatment with siRNA against Rap1A and/or B. In a flow chamber model with pre-established human umbilical vein endothelial cells (HUVECs), Rap1A/B downregulated MSCs interacted for longer distances before arrest, indicating adhesion defects. CXCL12-induced adhesion of MSCs on immobilized Vascular Cell Adhesion Molecule (VCAM)-1 was also decreased after the downregulation of Rap1A, Rap1B, or both, as was CXCL12-induced transwell migration. In a competitive murine short-term homing model with i.v. co-injection of Rap1A+B siRNA-treated and control MSCs that were labeled with PKH 26 and PKH 67 fluorescent dyes, the Rap1A+B siRNA-treated MSCs were detected at increased frequencies in blood, liver, and spleen compared to control MSCs. Thus, Rap1 GTPase modulates the adhesion and migration of MSCs in vitro and may increase the bio-availability of i.v.-transplanted MSCs in tissues in a murine model.

Background: The management of blood supply depends, among other factors, on the effective remobilization of first‐time donors (FTDs). This study investigates the efficacy of telephone calls to increase second donation rates.Study Design and Methods: A randomized controlled trial was conducted on 418 first‐time blood donors. In the telephone group (TG, n = 206), men were contacted 9–10 and women 13–14 weeks after their first donation. They were asked about satisfaction and intention to return, and offered an appointment. The primary outcome was the return rate within 6 months after the first donation.Results: The mean age was 28.8 ± 10.0 years and 59.9% of FTDs were female. In the TG, 89.3% were reached. Approximately 50% of each group had donated a second time by 24.2 weeks for the control group (CG) and 14.8 weeks for the TG. The six‐month return rate was 65.0% in the TG and 54.3% in the CG (95%‐CI [0.9%; 20.6%]; p = .033). The restricted mean time to return within 6 months was 19.4 weeks in the CG compared to 17.2 weeks in the TG (95%‐CI [0.7; 3.7]; p = .004). The intervention effect tended to be larger in men than in women.Discussion: Contacting FTDs by phone after their first donation increases the six‐month return rate and reduces the interval to a second donation. Male donors appear to be more receptive to this intervention. Whether the effect of the intervention helps to establish a donor identity in the long term should be the subject of further studies.

Introduction: Reporting of transfusion reactions is good practice and required by many guidelines. Errors in the transfusion chain can also lead to severe patient reactions and depend on active error reporting. We aimed to characterize transfusion incidents and asked whether workup of transfusion reactions may also contribute to revealing logistical errors. Methods: Transfusion medical records from 2011 to 2019 at our tertiary medical centre, as well as forensic autopsy reports, digitized sections, and court records from 1990 to 2019 were analysed. A total of 230,845 components were transfused between 2011 and 2019 at our own institution. Results: Overall, 322 transfusion incidents were reported. Of these, 279 were from our own institution, corresponding to a frequency of 0.12% of all transfusions. The distribution of reaction types is consistent with the literature, with allergic reactions (55.9%), febrile-non-hemolytic reactions (FNHTR, 24.2%), hemolytic reactions (3.4%) and other types at smaller frequencies (< 3%). Twenty-nine (10.4%) of the 279 reports revealed logistical errors, including hemoglobin above guideline threshold (4.3%), incorrect or non-performed bedside tests (3.2%), inadequate patient identification (2.5%), laboratory and issuing errors, missed product checks or failure to follow recommendations (1.1% each). Eight of 29 (27.5%) of the logistical errors were detected by serendipity during workup of incident reports. In addition, 8/932 autopsy cases under code A14 (medical treatment errors) were found to be transfusion-associated (0.9%). Conclusion: Systematic workup of transfusion incidents can identify previously undetected errors in the transfusion chain. Passive reporting of errors through the recording of side effects may serve as a tool to assess more closely assess the frequency and quality of handling errors in real life, and thus serve to improve patient safety.

Background and Objectives: Serum eye drops (SEDs) are used to treat ocular surface disease (OSD) and to promote ocular surface renewal. However, their use and production are not standardized, and several new forms of human eye drops have been developed.Materials and Methods: The International Society for Blood Transfusion Working Party (ISBT WP) for Cellular Therapies held a workshop to review the current types of eye drops of human origin (EDHO) status and provide guidance.Results: The ISBT WP for Cellular Therapies introduced the new terminology ‘EDHO’ to emphasize that these products are analogous to ‘medical products of human origin’. This concept encompasses their source (serum, platelet lysate, and cord blood) and the increasingly diverse spectrum of clinical usage in ophthalmology and the need for traceability. The workshop identified the wide variability in EDHO manufacturing, lack of harmonized quality and production standards, distribution issues, reimbursement schemes and regulations. EDHO use and efficacy is established for the treatment of OSD, especially for those refractory to conventional treatments.Conclusion: Production and distribution of single‐donor donations are cumbersome and complex. The workshop participants agreed that allogeneic EDHO have advantages over autologous EDHO although more data on clinical efficacy and safety are needed. Allogeneic EDHOs enable more efficient production and, when pooled, can provide enhanced standardization for clinical consistency, provided optimal margin of virus safety is ensured. Newer products, including platelet‐lysate‐ and cord‐blood‐derived EDHO, show promise and benefits over SED, but their safety and efficacy are yet to be fully established. This workshop highlighted the need for harmonization of EDHO standards and guidelines.