抄録
We previously reported that a novel continuous cell separation method using a seal-less continuous flow centrifuge was capable of separating a large number of nucleated cells with minimum damage during only a few hours of operation. Under a centrifugal force field, the introduction of cell suspension into the proximal portion of the channel resulted in continuous separation of cells according to their densities. The performance of the apparatus was demonstrated on the separation of human buffy coat. As for the differential leukocyte count on smear preparation with May-Grünwald-Giemsa stain, Lymphocytes were mainly present in fraction 4 (d=1.070) at over 90% without red blood cells. Neutrophils were observed in fraction 6 (d=1.080) at more than 90%, but red blood cells were included. Lymphocytes and neutrophils were mostly well separated. In fraction 5 (d=1.075), basophils were concentrated at near 15%. On a result analyzed by flow cytometry, lymphocytes (CD45+, CD14-) were mostly collected in fraction 3 and 4, granulocytes (CD45±, CD14-), which mainly consisted of neutrophils, were collected in fraction 5 and 6. Monocytes (CD45+, CD14+) and CD34 positive cells were collected in fraction 2. These results indicated that the present method was capable of concentrating rare cells such as basophils or CD34 positive cells in a proper fraction. The method may be applied to concentrate hematopoietic stem cells. [Jpn J Physiol 54 Suppl:S113 (2004)]
