PubMed · 11347098
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M Washio, T Yanase, H Tanaka. 2001. [Primary hypercholesterolemia].. https://pubmed.ncbi.nlm.nih.gov/11347098/
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Dendritic cells (DC) are increasingly used for the immunotherapy of cancer. Both the induction of tumor-specific T cells and some clinical regressions have been observed in early phase I/II trials by using either DC isolated from blood, DC generated from CD34+ precursors ex vivo, and most frequently, by employing monocyte-derived DC. As DC vaccination is now awaiting phase II/III trials with larger patient collectives, it becomes increasingly important to overcome prior limitations such as the repetitive, labor-intensive generation of DC in a large number of open culture vessels. We describe here as a result of several years of optimization, in detail, a procedure that uses the so-called Nunc cell factories to process a whole apheresis product, labor- and cost-effectively in a quasi-closed system to reproducibly generate (by using GM-CSF+IL-4 followed by a maturation cocktail composed of IL-1beta+IL-6+TNF-alpha +PGE(2)) large numbers (8.32+/-3.8% of input peripheral blood mononuclear cells (PBMC)) of mature (>85% CD83+), monocyte-derived DC that can be successfully cryopreserved. Our report is based on the processing of >100 aphereses including 52 unselected aphereses in advanced melanoma patients. This allows us also to suggest meaningful quality and validation criteria. The DC generation method appears particularly promising as respective DC vaccination proved to be immunogenic in cancer patients and cell factories can readily be converted to a fully closed system by using appropriate valves, tubings, and bags.
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Multicomponent apheresis procedures offer the possibility to collect standardized blood components as compared to whole blood donations. A new program for the concurrent collection of platelets (PLTs) and red blood cells (RBCs) was evaluated in a prospective study. Apheresis donors ( n=18) underwent concurrent collection of PLTs and RBCs using the Haemonetics MCS+ blood cell separator. Aliquots of PLTs and RBCs were collected during five to six passes of the discontinuous flow procedure. The platelet product was in-line filtered during the last pass of the separation procedure. After collection, saline-adenine-glucose-mannitol (SAGM) preservative solution was automatically added to the RBCs. Thereafter, the RBCs were in-line leukodepleted by gravity filtration at room temperature. The PLTs and RBCs were subsequently stored at 22+/-2 degrees C for 5 days and 4+/-2 degrees C for 35 days, respectively. The following in vitro parameters were evaluated over the storage periods: blood cell counts, glucose, lactate, lactate dehydrogenase, pH, plasma hemoglobin, and potassium. Two ready-to-use blood components from one donor were collected in an average procedure time of 86+/-10 min; 2.47+/-0.74 x 10(11) PLTs were collected in a product volume of 232+/-43 ml. The RBC volume averaged 280+/-20 ml and the hemoglobin content was 56.8+/-2.4 g per unit. The leukocyte contamination of the platelet product was 0.44+/-0.56 x 10(5) and the residual leukocyte content of the RBC product was 0.28+/-0.02 x 10(5). Storage data showed no relevant drop in pH. Day 35 results of the RBC products showed that all of the units had less than 0.8% hemolysis. Standardized PLT and RBC products of good quality can be concurrently collected with the MCS+ blood cell separator. In vitro testing of the products collected and stored for 5 and 35 days, respectively, met the Council of Europe criteria for leukodepleted blood products.