N Sasani1, R Roghanian1, G Emtiazi1, S M Jalali2, M Nikougoftar Zarif3, A Aghaie4. 1. Department of Cell and Molecular Biology & Microbiology, Faculty of Biological Science and Technology, University of Isfahan, 81746-79441 Isfahan, Iran. 2. IBRF-1 plasma collection center, Iranian Blood Research and Fractionation Company, Fardis, Alborz, Iran. 3. Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, 14665-1157 Tehran, Iran. 4. Blood Transfusion Research Center, High Institute for Research and Education in Transfusion Medicine, 14665-1157 Tehran, Iran. Electronic address: Aghaie.a@gmail.com.
Abstract
OBJECTIVES: The use of leukoreduction filters has been highly increased in Iranian Blood Transfusion Centers within the last decade to provide sufficient leukoreduced blood products from healthy repeated donors for alloimmunized or sensitive recipients. Leucoflex LCR5, the dominant brand which procured by the Iranian Blood Transfusion Organization, is the most updated generation of the filters used around the world. MATERIAL AND METHODS: In this study, we recovered trapped leukocytes from these filters using different buffer solutions and optimized elution method. The count of recovered cells assessed by cell counter, and cell viability was detected using trypan blue staining. The percent of leukocyte subpopulations was evaluated using a panel of monoclonal antibodies and flow cytometric analysis. RESULTS: It illustrated that a buffer solution consistent with PBS in pH 7.2 containing 2mM EDTA and 4% (w/w) Dextran 40 was the best buffer for LCR5 filter backflushing. The white cell counted as 4.56×108 Granulocytes, 3.34×108 Lymphocytes, and 0.64×108 Monocytes according to analysis with auto hemoanalysis and flow cytometric methods. CONCLUSION: The study guides and assists blood management systems in arranging a national blood profile database for future cell therapy strategies. Also, the recovered cells could be of significance in stem cell research, cellular interaction studies as well as novel molecular developments in drug discovery.
OBJECTIVES: The use of leukoreduction filters has been highly increased in Iranian Blood Transfusion Centers within the last decade to provide sufficient leukoreduced blood products from healthy repeated donors for alloimmunized or sensitive recipients. Leucoflex LCR5, the dominant brand which procured by the Iranian Blood Transfusion Organization, is the most updated generation of the filters used around the world. MATERIAL AND METHODS: In this study, we recovered trapped leukocytes from these filters using different buffer solutions and optimized elution method. The count of recovered cells assessed by cell counter, and cell viability was detected using trypan blue staining. The percent of leukocyte subpopulations was evaluated using a panel of monoclonal antibodies and flow cytometric analysis. RESULTS: It illustrated that a buffer solution consistent with PBS in pH 7.2 containing 2mM EDTA and 4% (w/w) Dextran 40 was the best buffer for LCR5 filter backflushing. The white cell counted as 4.56×108 Granulocytes, 3.34×108 Lymphocytes, and 0.64×108 Monocytes according to analysis with auto hemoanalysis and flow cytometric methods. CONCLUSION: The study guides and assists blood management systems in arranging a national blood profile database for future cell therapy strategies. Also, the recovered cells could be of significance in stem cell research, cellular interaction studies as well as novel molecular developments in drug discovery.