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Enrichment of normal progenitors in counter-flow centrifugal elutriation (CCE) fractions of fresh chronic myeloid leukemia leukapheresis products

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Dlubek D, Dybko J, Wysoczanska B, Laba A, Klimczak A, Kryczek I, Konopka L, Lange A. Enrichment of normal progenitors in counter-flow centrifugal elutriation (CCE) fractions of fresh chronic myeloid leukemia leukapheresis products.

Eur J Haematol 2002: 68: 281–288. © Blackwell Munksgaard 2002. Abstract:

Objectives: The aim of this study was to assess the suitability of a technique based on counter-flow centrifugal elutriation (CCE), which should allow one to enrich chronic myeloid leukemia (CML) patients' unstimulated native leukapheresis product (nLP) in CD34+ HLADR cells and BCR-ABL negative cells. Methods: Six newly diagnosed CML patients were subjected to leukapheresis, and the products were subfractionated with the use of CCE. nLP and all fractions were studied for the presence of CD34+ cells and a proportion of BCR-ABL fluorescence in situ hybridization (FISH)+ cells. Results: CCE fractions with a high flow rate contained the highest proportion of CD34+ cells [mean (SEM) 6.89% (3.88)]. However, CD34+ cells present in low-rate CCE fractions showed a higher proportion of HLADR[49.6% (13.5 in 70 mL min−1) and 21.5% (11.6 in 110 mL min−1)] than those in 170 mL min−1[3.2% (2.5)] and ‘rotor off’[3.4% (1.9)]. This was associated with lower proportions of BCR-ABL FISH+[8.1% (4.8) and 1.9 (1.7)] and smaller BCR-ABL to ABL transcript ratios [0.58 ( 17) and 0.26 (0.08) in 70 and 110 mL min−1] fractions as compared to 140 and 170 mL min−1 fractions [21.6% (5.2) and 31.6% (15.3) for BCR-ABL FISH+ cells and 0.75 (0.16) and 0.90 (0.24) for BCR-ABL/ABL]. Fractions with the lowest proportions of BCR-ABL-positive cells and the lowest BCR-ABL/ABL transcript ratios (110 mL min−1) contained from 1.3 × 106 to 82.7 × 106 (median: 3.97 × 106) CD34+ cells. Conclusions: In the present study we have shown that CCE may be used effectively to obtain nLP fractions enriched in normal hematopoietic progenitors.
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Keywords: CD34 BCR-ABL-negative; chronic myeloid leukemia; counter-flow centrifugal elutriation

Document Type: Research Article

Affiliations: 1: Lower Silesian Centre for Cellular Transplantation and Institute of Immunology and Experimental Therapy, Wroclaw, Poland; 2: Department of Internal Disease, Institute of Hematology and Transfusiology, Warszawa, Poland

Publication date: May 1, 2002

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