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Combined negative and positive selection of mobilized CD34 + blood cells
Author(s) -
Di Nicola Massimo,
Bregni Marco,
Siena Salvatore,
Adelchi Ruffini Pier,
Milanesi Marco,
Ravagnani Fernando,
Massimo Gianni A.
Publication year - 1996
Publication title -
british journal of haematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.907
H-Index - 186
eISSN - 1365-2141
pISSN - 0007-1048
DOI - 10.1046/j.1365-2141.1996.00725.x
Subject(s) - percoll , cd34 , chemistry , ficoll , andrology , microbiology and biotechnology , immunology , centrifugation , biology , chromatography , stem cell , biochemistry , medicine , peripheral blood mononuclear cell , in vitro
We tested four negative and two positive selection methods for separation of CD34 + cells from mobilized blood cells, and analysed fold‐enrichment, purity and recovery of CD34 + cells after selection procedures. The elimination of mature CD34 − cells was achieved by adhesion to nylon‐wool fibre (5.9 ± 1.0 mean fold‐enrichment and 65.2 ± 2.3 mean recovery of CD34 + cells). Standard or modified Ficoll‐Hypaque and Percoll density gradients, as well as phagocytosis with magnetic beads, were less effective in eliminating CD34 − cells, both purity and fold‐enrichment of CD34 + cells being lower than those obtained with separation by nylon‐wool. Both positive selection methods tested, Ceprate and MiniMacs System, generated highly purified CD34 + cell populations ranging from 80% to 90%. The recovery of CD34 + cells was optimal with MiniMacs (77.9±3.6) and low with Ceprate (28.8±2.8). Based on these results, in two large‐scale experiments we combined nylon‐wool fibre and MiniMacs System in a two‐step separation procedure obtaining a 36.9±2.6 mean fold‐enrichment and a 50.5±0.3 mean recovery of CD34 + cells. In this way we achieved optimal enrichment and recovery of CD34 + cells, with a substantial saving of cost compared to either selection method alone.