title: Pre-clinical development of cord blood-derived progenitor cell graft expanded ex vivo with cytokines and the polyamine copper chelator tetraethylenepentamine.
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, purified 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.
  
  
  CONCLUSIONS
  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.
authors.affiliation: Gamida-Cell Ltd, Jerusalem, Israel.
authors.forename: T
authors.initials: T
authors.lastname: Peled
authors.forename: J
authors.initials: J
authors.lastname: Mandel
authors.forename: R N
authors.initials: RN
authors.lastname: Goudsmid
authors.forename: C
authors.initials: C
authors.lastname: Landor
authors.forename: N
authors.initials: N
authors.lastname: Hasson
authors.forename: D
authors.initials: D
authors.lastname: Harati
authors.forename: M
authors.initials: M
authors.lastname: Austin
authors.forename: A
authors.initials: A
authors.lastname: Hasson
authors.forename: E
authors.initials: E
authors.lastname: Fibach
authors.forename: E J
authors.initials: EJ
authors.lastname: Shpall
authors.forename: A
authors.initials: A
authors.lastname: Nagler
mesh_terms: D000071916:AC133 Antigen; D015703:Antigens, CD / Q000378:metabolism; D018952:Antigens, CD34 / Q000378:metabolism; D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D002614:Chelating Agents / Q000494:pharmacology*; D016207:Cytokines / Q000494:pharmacology*; D005029:Ethylenediamines / Q000494:pharmacology*; D005312:Fetal Blood / Q000166:cytology*; D006023:Glycoproteins / Q000378:metabolism; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D007231:Infant, Newborn; D010455:Peptides / Q000378:metabolism
chemical_list: D000071916:AC133 Antigen; D015703:Antigens, CD; D018952:Antigens, CD34; D002614:Chelating Agents; D016207:Cytokines; D005029:Ethylenediamines; D006023:Glycoproteins; C000606023:PROM1 protein, human; D010455:Peptides; C000606024:Prom1 protein, mouse; C034269:tetraethylenepentamine