Sterility release testing of peripheral blood stem cells for transplantation: impact of culture bottles and incubation temperature.

BACKGROUND: Sterility testing of peripheral blood stem cells (PBSCs) is mandatory before release. As antibiotic treatment of the PBSC donor may result in false-negative results, PBSC matrix validation must be carried out. STUDY DESIGN AND METHODS: Three spiked PBSCs and a buffy coat (BC; control matrix) were analyzed using the blood culture device BacT/ALERT 3D with the low-temperature module. Samples were spiked with Staphylococcus aureus, Bacillus subtilis, Pseudomonas aeruginosa, Candida albicans, Aspergillus brasiliensis, Clostridium sporogenes, and Propionibacterium acnes. Standard iAST/iNST culture bottles and iFA/iFN Plus bottles, which include resorbing polymers, were incubated for 14 days. All aerobic bottles were incubated at 22.5°C and for a direct comparison also at 35°C while all anaerobic bottles were incubated at 35°C. RESULTS: The BacT/ALERT 3D system detected all microbes in iAST/iNST culture bottles according to their growth behavior in the BC matrix. Detection of microbes differed significantly in PBSC products using standard iAST/iNST culture bottles and iFA/iFN Plus bottles with resorbing polymers: In Graft 1 no growth was detected in spiked bottles with S. aureus (iAST), B. subtilis (iAST/iNST), C. sporogenes (iNST), and P. acnes (iNST) compared to iFA Plus and iFN Plus bottles wherein growth of spiked microbes was confirmed. Graft 2, with another antibiotic treatment, showed no growth in iAST/iNST bottles spiked with P. aeruginosa, B. subtilis, and C. sporogenes. However, using iFA/iFN Plus bottles all spiked microbes were detectable. The comparison of incubation temperature showed an expected slower growth at 22.5°C. CONCLUSION: The use of iFA/iFN Plus culture bottles incubated at different temperatures safely detected microbes in spiked PBSCs.

Quality of irradiated and non-irradiated plateletpheresis concentrates stored in platelet additive solution.

INTRODUCTION: Platelet additive solutions (PAS) allow to maintain platelet storage properties in platelet concentrates (PCs). The aim of the present study was to evaluate the in-vitro quality of irradiated and non-irradiated PCs, suspended in PAS, over a storage period of 6 days. METHODS: Plateletpheresis donors fulfilling current eligibility criteria underwent plateletpheresis with the MCS+ blood cell separator. The PAS SSP+ was used to store platelets (PLT) for up to 6 days. Aliquots were drawn from the PCs after collection, at day 4, 5 and 6 of storage. A battery of tests was performed to analyse the quality of the PCs: PLT count, mean PLT volume (MPV), PLT activation marker CD 62, swirl, RBC and WBC contamination, pH, citrate, glucose, lactate and lactate dehydrogenase. RESULTS: An average of 2.53 ±â€¯0.21 × 1011 PLT were collected in a product volume of 231 ±â€¯5 mL in irradiated and 233 ±â€¯6 mL in non-irradiated PCs, respectively. RBC- and WBC-contamination were within the allowed ranges. Δ CD62 steadily decreased in irradiated and non-irradiated PCs while the pH was well maintained over storage time. Glucose and lactate levels of irradiated and non-irradiated PCs showed characteristic pattern of PC storage within acceptable ranges. CONCLUSION: Our data demonstrate that parameters of PC quality were well maintained over a storage period of 6 days using PAS. Irradiation had no impact on the quality of PCs. The product quality of irradiated and non-irradiated PCs met national and European guidelines.

Immunoglobulin G levels during collection of large volume plasma for fractionation.

BACKGROUND: There is a need of comprehensive work dealing with the quality of plasma for fractionation with respect to the IgG content as today most plasma derivates are used to treat patients with immunodeficiencies and autoimmune disorders. Therefore, a prospective study was carried out to analyse IgG levels before plasmapheresis and every 200ml collected plasma. MATERIALS AND METHODS: Fifty-four experienced plasmapheresis donors were recruited for subsequent 850ml plasmapheresis using the Aurora Plasmapheresis System. Donors peripheral blood counts were analysed before and after plasmapheresis using an electronic counter. Total protein, IgG and citrate were measured turbidometrically before, during and after apheresis as well as in the plasma product. Furthermore, platelets, red and white blood cells were analysed as parameters of product quality. RESULTS: An average of 2751±247ml blood was processed in 47±6min. The collected plasma volume was 850±1mL and citrate consumption was 177±15mL. A continuous drop of donors' IgG level was observed during plasmapheresis. The drop was 13% of the IgG baseline value at 800mL collected plasma. Total protein, IgG and cell counts of the plasma product met current guidelines of plasma for fractionation. CONCLUSION: Donors' IgG levels during apheresis showed a steady decrease without compromising the quality of plasma product.

Detection of the relatively slow-growing Propionibacterium acnes in seven matrices of blood components and advanced therapeutical medicinal products.

BACKGROUND: Relatively slow-growing bacteria like Propionibacterium acnes represent a challenge for quality control investigations in sterility release testing of blood components and advanced therapeutic medicinal products (ATMPs). METHODS: A convenient validation with 7 matrices was performed using buffy coat, stem cells, islet cells, natural killer cells, red blood cells, platelets and plasma in the microbial detection system Bact/Alert®3D incubator. All matrix samples were spiked twofold with Propionibacterium acnes with approximately 50 colony forming units (CFUs) per bottle in iAST and iNST culture bottles for 14days using a multishot bioball. Additionally, the stem cell preparations were also incubated in iFAplus and iFNplus culture bottles, which include neutralizing polymers. RESULTS: The Bact/Alert®3D-System detected Propionibacterium acnes in anaerobic culture bottles in buffy coat [3.3 d (=positive signal day to detection as mean value)], red blood cells [3.2 d], platelets [3.3], plasma [3.7 d], natural killer cells [3.3 d] and islet cells [4.9 d], resp. No growth of Propionibacterium was found in autologous stem cells using iAST and iNST culture bottles. However, Propionibacterium was safely detected in the iFNplus culture bottle with polymers in the stem cell matrix. A successful validation of media was performed. CONCLUSIONS: Our study shows that Bact/Alert®3D-System safely detects the relatively slow-growing bacterium Propionibacterium acnes in different matrices in a practical way except stem cells. Using the iFNplus culture bottle for stem cell products positive signals were observed.