TY - JOUR
T1 - Pre-clinical development of cord blood-derived progenitor cell graft expanded ex vivo with cytokines and the polyamine copper chelator tetraethylenepentamine
AU - Peled, T.
AU - Mandel, J.
AU - Goudsmid, R. N.
AU - Landor, C.
AU - Hasson, N.
AU - Harati, D.
AU - Austin, M.
AU - Hasson, A.
AU - Fibach, E.
AU - Shpall, E. J.
AU - Nagler, Arnon
N1 - Copyright:
Copyright 2017 Elsevier B.V., All rights reserved.
PY - 2004
Y1 - 2004
N2 - 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.
AB - 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.
KW - Ex vivo large-scale expansion
KW - Pre-clinical development
KW - Tetraethylenepentamine
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U2 - 10.1080/14653240410004916
DO - 10.1080/14653240410004916
M3 - Article
C2 - 16146887
AN - SCOPUS:4544362434
SN - 1465-3249
VL - 6
SP - 344
EP - 355
JO - Cytotherapy
JF - Cytotherapy
IS - 4
ER -