METHOD FOR CALCULATING THE DOSE OF HAEMATOPOIESIS PRECURSOR CELLS IN A LEUKOCYTAPHERESIS PRODUCT BY TAKING INTO ACCOUNT THE CHANGE IN THE INTEGRITY OF CELL MEMBRANES DURING STORAGE

摘要

FIELD: medicine.;SUBSTANCE: invention refers to medicine, namely immunology, and can be used for calculating transplantation dose of CD34-positive progenitor cells in a donor leukocytapheresis product. Calculation method involves labeling cell material with fluorochrome-conjugated monoclonal antibodies to CD34 and CD45 markers with simultaneous staining with fluorescent DNA-tropic dye 7-aminoactinomycin D. Further, CD34-positive haematopoiesis progenitors contents are determined by haemocytometry and laser flow cytofluorometry. When calculating a transplantation dose, factors of membrane integrity change of haemopoietic stem cells – 0.985, 0.971, 0.958, 0.947 and 0.930, respectively, for 1, 2, 3, 4 and 5^th days storage of cell material before transplantation to recipient at temperature plus 2 – plus 6 °C. Calculation is carried out by formulas 1 and 2:; ,; ,;where CD34⁺ rel. – relative content of CD34⁺ progenitor cells of haematopoiesis in a leukocytapheresis product; CD34 abs. is number of progenitor cells, which is defined as the number of events with high CD34 antigen expression, low CD45 antigen expression and low granularity, calculated in the polygonal region "HSC 1"; k_n is the CD34⁺ cell membrane integrity change coefficient on the n-th storage day; CD45 abs. – number of leukocytes in the sample, which is determined as the number of CD45 antigen-positive events counted in the rectangular region "CD45⁺"; Dose of CD34⁺ – transplant dose of CD34⁺ haematopoiesis progenitor cells in leukocytapheresis product; C (leuk.) – concentration of leukocytes in sample, cells/mcl; V – volume of leukocytapheresis product, mcl; m is body weight of recipient of CD34-positive hematopoiesis progenitor cells, kg.;EFFECT: use of the given method allows calculating dose of CD34⁺ cells considering change of integrity of their membranes during storage in conditions of cooling before transplantation.;1 cl, 7 dwg, 1 tbl, 2 ex