Combined negative and positive selection of mobilized CD34+ blood cells

Massimo Di Nicola, Marco Bregni, Salvatore Siena, Pier Adelchi Ruffini, Marco Milanesi, Fernando Ravagnani, A. Massimo Gianni

Research output: Contribution to journalArticle

Abstract

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 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.

Original languageEnglish
Pages (from-to)716-721
Number of pages6
JournalBritish Journal of Haematology
Volume94
Issue number4
Publication statusPublished - 1996

Keywords

  • CD34 cell separation
  • Mini-Macs System
  • Nylon-wool fibre

ASJC Scopus subject areas

  • Hematology

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    Di Nicola, M., Bregni, M., Siena, S., Ruffini, P. A., Milanesi, M., Ravagnani, F., & Gianni, A. M. (1996). Combined negative and positive selection of mobilized CD34+ blood cells. British Journal of Haematology, 94(4), 716-721.