Clinical-scale validation of a new efficient procedure for cryopreservation of ex vivo expanded cord blood hematopoietic stem and progenitor cells.
Survival of ex vivo expanded hematopoietic stem cells (HSC) and progenitor cells is low with the standard cryopreservation procedure. We recently showed that the efficiency of cryopreservation of these cells may be greatly enhanced by adding a serum-free xeno-free culture medium (HP01 Macopharma), which improves the antioxidant and biochemical properties of the cryopreservation solution. Here we present the clinical-scale validation of this cryopreservation procedure. The hematopoietic cells expanded in clinical-scale cultures were cryopreserved applying the new HP01-based procedure. The viability, apoptosis rate and number of functional committed progenitors (methyl-cellulose colony forming cell test), short-term repopulating HSCs (primary recipient NSG mice) and long-term HSCs (secondary recipient NSG mice) were tested before and after thawing. The efficiency of clinical-scale procedure reproduced the efficiency of cryopreservation obtained earlier in miniature sample experiments. Furthermore, the full preservation of short- and long-term HSCs was obtained in clinical scale conditions. Because the results obtained in clinical-scale volume are comparable to our earlier results in miniature-scale cultures, the clinical-scale procedure should be considered validated. It allows cryopreservation of the whole ex vivo expanded culture content, conserving full short- and long-term HSC activity.
Etablissement Français du Sang Aquitaine-Limousin, Bordeaux, France; U1035 INSERM/Université de Bordeaux, France.
Pascale
P
Duchez
Etablissement Français du Sang Aquitaine-Limousin, Bordeaux, France; U1035 INSERM/Université de Bordeaux, France.
Laura
L
Rodriguez
Etablissement Français du Sang Aquitaine-Limousin, Bordeaux, France; U1035 INSERM/Université de Bordeaux, France.
Jean
J
Chevaleyre
Etablissement Français du Sang Aquitaine-Limousin, Bordeaux, France; U1035 INSERM/Université de Bordeaux, France.
Philippe Brunet
PB
De La Grange
Etablissement Français du Sang Aquitaine-Limousin, Bordeaux, France; U1035 INSERM/Université de Bordeaux, France. Electronic address: zoran.ivanovic@efs.sante.fr.
Zoran
Z
Ivanovic
D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D002478:Cells, Cultured; D015925:Cryopreservation / Q000379:methods*; D003451:Cryoprotective Agents / Q000494:pharmacology*; D003470:Culture Media / Q000494:pharmacology*; D005312:Fetal Blood / Q000166:cytology*; D018380:Hematopoietic Stem Cell Transplantation / Q000379:methods*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D014183:Transplantation, Heterologous
D003451:Cryoprotective Agents; D003470:Culture Media