[Blood fractionation based on the extraction of the buffy-coat layer. Analysis of our results]

To compare a procedure of blood processing via a quadruple bag for the preparation of white-cell-poor blood components with the results obtained with triple-bag-system, in order to adopt it as routine in our blood centre.

Blood was collected in 289 quadruple-bag-system containing 63 mL of CPD as anticoagulant and 100 mL of SAG M solution as additive for the red cells. We used 237 standard quadruple-bags supplied by Fenwal (195) and NPBI (42), and 52 of the top-and-bottom system supplied by Fenwal. Blood separation was made automatically by CompomatR (NPBI) and platelet concentrates were prepared from the buffy-coat fraction. Standard bags were processing as follows: After the first centrifugation of the whole blood (28,800 g), the plasma was transferred into the 300-mL bag until the interface of red cells and plasma was detected; then approximately 80 mL of plasma and buffy-coat (BC) were collected into the 100-mL satellite bag. Top-and-bottom bags were centrifuged at 43,500 g., the red-cells were transferred into the bottom-bag containing SAGM, and plasma was transferred into the top-bag. The buffy-coat fraction remains in the original bag. In both procedures, platelet concentrates were prepared from buffy-coat fraction. After the centrifugation of this fraction (1,400 g and 1,600 g), the supernatant (concentrated platelets in plasma) was transferred into a 300-mL bag and the bag with the residual buffy-coat was then discarded. 287 triple-bags were separated in the traditional way, using platelet-rich plasma (PRP) as a source for the preparation of platelet concentrate (PC). Volumes were measured by weigh and specificity gravity. Platelet, leukocytes and red-cells were counted in the Coulter-Counter (STKR.Izasa). T-Student test and Chi2 test were used for statistical analysis and p < 0.05 was taken as a significant difference between samples.

The three kinds of quadruple-bags showed results very homogeneous with little differences. For all brand of bags packed-red-cells showed a volume of 300 +/- 2.9 mL, EVF of 51.9 +/- 0.7%. The recovery of red cells into the packed red cells and of platelets into the platelet concentrate was 90 +/- 0.4 percent and 69 +/- 2 percent respectively, of the original value. White cells in the packed red cells were 9.7 +/- 03 x 10(9) with recovery of 30.1 +/- 1.4 percent of the original value; statistical difference was found in comparison with triple bags PRC (p < 0.001). The PC volumes averaged 71 +/- 1 mL and the overall mean platelet concentration was 77 +/- 2 x 10(9). Eighty three percent of PC contained more than 55 x 10(9). White cells contamination of platelet concentrates was 0.283 +/- 0.039 x 10(9), with a recovery of 9.5 +/- 1.5 percent of the leukocytes present in the whole blood. This value is below the threshold that prevents febrile reactions and microaggregate formation. The plasma yield in the quadruple-bag system was much greater than that in the triple-bag system (p < 0.001). With this process we removed about 77 +/- 0.5 percent of the plasma present in the original unit in comparison with 56 +/- 1 percent removed in that of the triple-bag system.

With this procedure leukocyte-poor blood components are obtained in which more of 90 percent original white cells have been removed, with a red-cell recovery of 90 percent. Moreover, the plasma yield is also excellent.