Pre-clinical development of cord blood-derived progenitor cell graft expanded ex vivo with cytokines and the polyamine copper chelator tetraethylenepentamine

T. Peled, J. Mandel, R. N. Goudsmid, C. Landor, N. Hasson, D. Harati, M. Austin, A. Hasson, E. Fibach, E. J. Shpall, Arnon Nagler

Research output: Contribution to journalArticlepeer-review

Abstract

Background. We have previously demonstrated that the copper chelator tetraethylenepentamine (TEPA) enables preferential expansion of early hematopoietic progenitor cells (CD34+ CD38-, CD34+ CD38- Lin-) in human umbilical cord blood (CB)-derived CD34+ cell cultures. This study extends our previous findings that copper chelation can modulate the balance between self-renewal and differentiation of hematopoietic progenitor cells. Methods. In the present study we established a clinically applicative protocol for large-scale ex vivo expansion of CB-derived progenitors. Briefly, CD133+ cells, purifed from CB using Miltenyi Biotec's (Bergisch Gladbach, Germany) CliniMACS separation device and the anti-CD133 reagent, were cultured for 3 weeks in a clinical-grade closed culture bag system, using the chelator-based technology in combination with early-acting cytokines (SCF, thrombopoietin, IL-6 and FLT-3 ligand). This protocol was evaluated using frozen units derived from accredited cord blood banks. Results. Following 3 weeks of expansion under large-scale culture conditions that were suitable for clinical manufacturing, the median output value of CD34+ cells increase by 89-fold, CD34+ CD38- increase by 30-fold and CFU cells (CFUc) by 172-fold over the input value. Transplantation into sublethally irradiated non-obese diabetic (NOD/SCID) mice indicated that the engraftment potential of the ex vivo expanded CD133+ cells was significantly superior to that of unexpanded cells: 60 ± 5.5% vs. 21 ± 3.5% CD45+ cells, P = 0.001, and 11 ± 1.8% vs. 4 ± 0.68% CD45+ CD34+ cells, P = 0.012 n = 32, respectively. Discussion. Based on these large-scale experiments, the chelator-based ex vivo expansion technology is currently being tested in a phase 1 clinical trial in patients undergoing CB transplantation for hematological malignancies.

Original languageEnglish
Pages (from-to)344-355
Number of pages12
JournalCytotherapy
Volume6
Issue number4
DOIs
StatePublished - 2004
Externally publishedYes

Keywords

  • Ex vivo large-scale expansion
  • Pre-clinical development
  • Tetraethylenepentamine

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