title: In vitro and in vivo evaluation of cord blood hematopoietic stem and progenitor cells amplified with glycosaminoglycan mimetic.
abstract: BACKGROUND
  Expansion protocols aim at both increasing the number of umbilical cord blood (UCB) hematopoietic stem cells and progenitor cells (HSPCs) and reducing the period of neutropenia in UCB HSPC graft. Because glycosaminoglycans (GAGs) are known to be important components of the hematopoietic niche and to modulate growth factor effects, we explored the use of GAG mimetic OTR4131 to potentiate HSPC's in vitro expansion and in vivo engraftment.
  
  
  METHODS
  UCB CD34+ cells were expanded with serum-free medium, SCF, TPO, FLT3-lig and G-CSF during 12 days in the absence or the presence of increasing OTR4131 concentrations (0-100 μg/mL). Proliferation ratio, cell viability and phenotype, functional assays, migration capacity and NOD-scid/γc(-/-) mice engraftment were assessed after expansion.
  
  
  RESULTS
  At Day 12, ratios of cell expansion were not significantly increased by OTR4131 treatment. Better total nucleated cell viability was observed with the use of 1 μg/mL GAG mimetic compared to control (89.6 % ± 3.7 % and 79.9 % ± 3.3 %, respectively). Phenotype analysis showed a decrease of monocyte lineage in the presence of OTR4131 and HSPC migration capacity was diminished when GAG mimetic was used at 10 μg/mL (10.9 % ± 4.1 % vs. 52.9 % ± 17.9 % for control). HSPC clonogenic capacities were similar whatever the culture conditions. Finally, in vivo experiments revealed that mice successfully engrafted in all conditions, even if some differences were observed during the first month. Three months after graft, bone marrow chimerism and blood subpopulations were similar in both groups.
  
  
  CONCLUSIONS
  UCB HSPCs ex-vivo expansion in the presence of OTR4131 is a safe approach that did not modify cell function and engraftment capacities. In our experimental conditions, the use of a GAG mimetic did not, however, allow increasing cell expansion or optimizing their in vivo engraftment.
authors.affiliation: Inserm, U 1160, Centre d'Investigation Clinique en Biothérapies, 75010, Paris, France. lionel.faivre@aphp.fr.|AP-HP, Hôpital Saint-Louis, Unité de Thérapie Cellulaire, Paris, F-75010, France. lionel.faivre@aphp.fr.
authors.forename: Lionel
authors.initials: L
authors.lastname: Faivre
authors.affiliation: Département d'Expérimentation d'Animale, Université́ Paris Diderot, Institut Universitaire d'Hématologie, Hôpital Saint-Louis, Paris, F-75010, France. veronique.parietti@gmail.com.
authors.forename: Véronique
authors.initials: V
authors.lastname: Parietti
authors.affiliation: OTR3 Company, 4 rue Française, 75001, Paris, France. fernando.sineriz@otr3.com.
authors.forename: Fernando
authors.initials: F
authors.lastname: Siñeriz
authors.affiliation: Université Paris Est Créteil, Université Paris Est, EA 4397 ERL CNRS 9215, Laboratoire CRRET, 61 Avenue du Général de Gaulle, 94010, Créteil, France. sandrine.chantepie@u-pec.fr.
authors.forename: Sandrine
authors.initials: S
authors.lastname: Chantepie
authors.affiliation: OTR3 Company, 4 rue Française, 75001, Paris, France. marie.gilbert@otr3.com.
authors.forename: Marie
authors.initials: M
authors.lastname: Gilbert-Sirieix
authors.affiliation: Université Paris Est Créteil, Université Paris Est, EA 4397 ERL CNRS 9215, Laboratoire CRRET, 61 Avenue du Général de Gaulle, 94010, Créteil, France. albanese@u-pec.fr.
authors.forename: Patricia
authors.initials: P
authors.lastname: Albanese
authors.affiliation: Inserm, U 1160, Centre d'Investigation Clinique en Biothérapies, 75010, Paris, France. jerome.larghero@aphp.fr.|AP-HP, Hôpital Saint-Louis, Unité de Thérapie Cellulaire, Paris, F-75010, France. jerome.larghero@aphp.fr.|Université Paris Diderot, Sorbonne Paris Cité, U 1160, Paris, F-75010, France. jerome.larghero@aphp.fr.
authors.forename: Jérôme
authors.initials: J
authors.lastname: Larghero
authors.affiliation: Inserm, U 1160, Centre d'Investigation Clinique en Biothérapies, 75010, Paris, France. valerie.vanneaux@aphp.fr.|AP-HP, Hôpital Saint-Louis, Unité de Thérapie Cellulaire, Paris, F-75010, France. valerie.vanneaux@aphp.fr.
authors.forename: Valérie
authors.initials: V
authors.lastname: Vanneaux
mesh_terms: D000818:Animals; D018929:Cell Culture Techniques; D049109:Cell Proliferation; D002478:Cells, Cultured; D005312:Fetal Blood / Q000166:cytology; D005936:Glucans / Q000494:pharmacology*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000502:physiology*; D006801:Humans; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D018716:Molecular Mimicry
chemical_list: D005936:Glucans; C542264:OTR 4131