Blood progenitor cell separation from clinical leukapheresis product by magnetic nanoparticle binding and magnetophoresis

Ying Jing, Lee R. Moore, P. Stephen Williams, Jeffrey J. Chalmers, Sherif S. Farag, Brian Bolwell, Maciej Zborowski

Research output: Contribution to journalArticle

88 Scopus citations

Abstract

Positive selection of CD34+ blood progenitor cells from circulation has been reported to improve patient recovery in applications of autologous transplantation. Current magnetic separation methods rely on cell capture and release on solid supports rather than sorting from flowing suspensions, which limits the range of therapeutic applications and the process scale up. We tested CD34+ cell immunomagnetic labeling and isolation from fresh leukocyte fraction of peripheral blood (leukapheresis) using the continuous quadrupole magnetic flow sorter (QMS), consisting of a flow channel (SHOT, Greenville, IN) and a quadrupole magnet with a maximum field intensity (Bo) of 1.42 T and a mean force field strength (Sm) of 1.45 x 108 TA/m 2. Both the sample magnetophoretic mobility (m) and the inlet and outlet flow patterns highly affect the QMS performance. Seven commercial progenitor cell labeling reagent combinations were quantitatively evaluated by measuring magnetophoretic mobility of a high CD34 expression cell line, KG-1a, using the cell tracking velocimeter (CTV). The CD34 Progenitor Cell Isolation Kit™ (Miltenyi Biotec, Bergisch Gladbach, Germany) showed the strongest labeling of KG-1a cells and was selected for progenitor cell enrichment from 11 fresh and 11 cryopreserved clinical leukapheresis samples derived from different donors. The CD34+ cells were isolated with a purity of 60-96%, a recovery of 18-60%, an enrichment rate of 12-169, and a throughput of (1.7-9.3) x 10 4 cells/s. The results also showed a highly regular dependence of the QMS performance on the flow conditions that agreed with the theoretical predictions based on the CD34+ cell magnetophoretic mobility.

Original languageEnglish (US)
Pages (from-to)1139-1154
Number of pages16
JournalBiotechnology and Bioengineering
Volume96
Issue number6
DOIs
StatePublished - Apr 15 2007
Externally publishedYes

Keywords

  • Cell separation
  • Flow sorting
  • Immunomagnetic labeling
  • Leukapheresis
  • Magnetophoresis
  • Progenitor cell

ASJC Scopus subject areas

  • Biotechnology
  • Bioengineering
  • Applied Microbiology and Biotechnology

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