Characterization of the transplant of the bone marrow cells by the level of its energy potential.

The goal of this study was to develop a methodfor rapid and adequate assessment for assessingthe quality of the graft bone marrow cells (BMC)on the change of their energy state using vitalfluorescent potential-sensitive probe iodide 2[p-(dimethylamino)styryl]-1-methylpyridinium(2-Di-1-ASP). Experimental studies were performed onsamples of native BMC suspension of 20 donorsin a standard solution of the stabilizer that werekept at room temperature. After mixing the cellsuspension were selected from it: the initial specimen and through 1, 2, 3, 4, 5, 24, 48, 72 - hoursstorage, stained with a probe 2-Di-1-ASP andexamined for the fluorescent microscope. Eachspecimen was measured by fluorescence 70-100single cells and mean fluorescence intensity ofBMC (F, arb.uints) was calculated. In each specimen was determined by the number of nucleatedcells (NC, %). Initially, the proportion of cells withintact cytoplasmic membranes in the specimenswere 80-92 % (by trypan blue test). The presence of mitoses in the samples of the suspensionafter 5 and 24 hours storage is shown in smearsof BMC, painted by the method Romanovsky. Statistical analysis of the dynamics of changes inF and NC in each sample was performed usinganalysis of variance dependent selections ANOVARepeated Measures. For all 20 donors was carried out by cluster analysis (Euclidean metric,strategy nearest middle). In all specimens, the BMC after 3 hours of storage at room temperature recorded F increasefrom baseline due to increase in the proportion ofcells with fluorescence intensity higher than 100arb.uints (N ); the degree of this increase correlates with an increase in NF>100. It is shown thatthe higher NF>100 3 hours after storage, the largerthe increase in the concentration of nucleatedcells in suspension, which started after 5 hoursstorage, being in accordance with revealed abilityof cells to division. With increasing NF>100 through3 hours storage BMC less than 1,66 times compared with the initial value, the concentrationof nucleated cells remained almost unchanged,indicating that the inability of cells to division inthese conditions. Developed and proposed a method for assessing the quality of the graft BMC of using the vitalfluorescent potential-sensitive probe 2-Di-1-ASPat room temperature in a solution of a standard stabilizer.