L Faivre1, M Chaussard2, L Vercellino3, V Vanneaux4, B Hosten5, K Teixera6, V Parietti7, P Merlet8, L Sarda-Mantel9, N Rizzo-Padoin10, J Larghero11. 1. Inserm, U 1160, centre d'investigation clinique en biothérapies, 75010 Paris, France; AP-HP, hôpital Saint-Louis, unité de thérapie cellulaire, 1, avenue Claude-Vellefaux, 75010 Paris, France. Electronic address: lionel.faivre@aphp.fr. 2. AP-HP, hôpital Saint-Louis, unité Claude-Kellershohn, 75010 Paris, France. Electronic address: michael.chaussard@gmail.com. 3. AP-HP, hôpital Saint-Louis, médecine nucléaire, 75010 Paris, France. Electronic address: laetitia.vercellino@aphp.fr. 4. Inserm, U 1160, centre d'investigation clinique en biothérapies, 75010 Paris, France; AP-HP, hôpital Saint-Louis, unité de thérapie cellulaire, 1, avenue Claude-Vellefaux, 75010 Paris, France. Electronic address: valerie.vanneaux@aphp.fr. 5. AP-HP, hôpital Saint-Louis, unité Claude-Kellershohn, 75010 Paris, France; Inserm, U1144, 75006 Paris, France; Université Paris-Descartes, faculté de pharmacie, 75006 Paris, France. Electronic address: benoit.hosten@aphp.fr. 6. AP-HP, hôpital Saint-Louis, unité Claude-Kellershohn, 75010 Paris, France. Electronic address: kellypteixeira@gmail.com. 7. Université Paris-Diderot, institut universitaire d'hématologie, hôpital Saint-Louis, département d'expérimentation d'animale, 75010 Paris, France. Electronic address: veronique.parietti@paris7.jussieu.fr. 8. AP-HP, hôpital Saint-Louis, médecine nucléaire, 75010 Paris, France; Université Paris-Diderot, Sorbonne-Paris-Cité, U 1160, 75010 Paris France. Electronic address: pascal.merlet@aphp.fr. 9. AP-HP, hôpital Saint-Louis, unité Claude-Kellershohn, 75010 Paris, France; AP-HP, hôpital Lariboisière, médecine nucléaire, 75010 Paris, France; Inserm UMR-S 942, hôpital Lariboisière, 75010 Paris, France. Electronic address: laure.sarda-mantel@aphp.fr. 10. AP-HP, hôpital Saint-Louis, unité Claude-Kellershohn, 75010 Paris, France; Inserm, U1144, 75006 Paris, France; Université Paris-Descartes, faculté de pharmacie, 75006 Paris, France. Electronic address: nathalie.rizzo@aphp.fr. 11. Inserm, U 1160, centre d'investigation clinique en biothérapies, 75010 Paris, France; AP-HP, hôpital Saint-Louis, unité de thérapie cellulaire, 1, avenue Claude-Vellefaux, 75010 Paris, France; Université Paris-Diderot, Sorbonne-Paris-Cité, U 1160, 75010 Paris France. Electronic address: jerome.larghero@aphp.fr.
Abstract
PURPOSE OF THE STUDY: After transplantation, cord blood (CB) hematopoietic stem and progenitor cells (HSPCs) are able to home to the bone marrow niche and to reconstitute the hematopoietic system. PET-CT imaging may be a useful method to monitor this parameter in different conditions. The aim of our study was to set up an efficient method for HSPC radiolabelling with [18F] fluorodeoxyglucose (18F-FDG) and to follow early HSPC homing through PET-CT in mice. MATERIALS AND METHODS: Purified CB HSPCs were radiolabelled with 18F-FDG at 37° C with various conditions of cell concentration, incubation time and radioactivity concentration in order to define the in vitro condition that allows both sufficient 18F-FDG uptake to get high quality PET imaging, and preservation of HSPC viability and functional properties during 3h after radiolabelling. Then, 24h after 2.25Gy irradiation, eight NOD-scid/γc-/- mice were injected with 18F-FDG-labelled HSPCs, the biodistribution of which was followed using micro-PET-CT. RESULTS: The optimal incubation time was 45min with a stability of 48.3%±12.8% after 180min. The radio-uptake rate we obtained was 7.2%±1.7% with an activity of 5.6±2.1 MBq. Three hours after radiolabelling, viability was 96.7%±3.4%. Fifteen hours after radiolabelling, cell viability was 64.0%±2.3%, migration ability diminished from 51.0%±23.6% to 12.0%±9.1%, clonogenic capacity was null, and long-term engraftment in NSG mice also decreased compared to unlabelled cells. Micro-PET-CT experiments showed an accumulation of radiolabelled HSPCs for 2.5h after injection in the bone marrow and a slight elution of 18F-FDG. CONCLUSION: The activity of the obtained 18F-FDG-labelled HSPCs was sufficient to perform the micro-PET-CT imaging. Although the radiolabelling had a significant toxicity on HSPCs 15h after labelling, this technique allowed monitoring the beginning of HSPC homing into the bone marrow.
PURPOSE OF THE STUDY: After transplantation, cord blood (CB) hematopoietic stem and progenitor cells (HSPCs) are able to home to the bone marrow niche and to reconstitute the hematopoietic system. PET-CT imaging may be a useful method to monitor this parameter in different conditions. The aim of our study was to set up an efficient method for HSPC radiolabelling with [18F] fluorodeoxyglucose (18F-FDG) and to follow early HSPC homing through PET-CT in mice. MATERIALS AND METHODS: Purified CB HSPCs were radiolabelled with 18F-FDG at 37° C with various conditions of cell concentration, incubation time and radioactivity concentration in order to define the in vitro condition that allows both sufficient 18F-FDG uptake to get high quality PET imaging, and preservation of HSPC viability and functional properties during 3h after radiolabelling. Then, 24h after 2.25Gy irradiation, eight NOD-scid/γc-/- mice were injected with 18F-FDG-labelled HSPCs, the biodistribution of which was followed using micro-PET-CT. RESULTS: The optimal incubation time was 45min with a stability of 48.3%±12.8% after 180min. The radio-uptake rate we obtained was 7.2%±1.7% with an activity of 5.6±2.1 MBq. Three hours after radiolabelling, viability was 96.7%±3.4%. Fifteen hours after radiolabelling, cell viability was 64.0%±2.3%, migration ability diminished from 51.0%±23.6% to 12.0%±9.1%, clonogenic capacity was null, and long-term engraftment in NSG mice also decreased compared to unlabelled cells. Micro-PET-CT experiments showed an accumulation of radiolabelled HSPCs for 2.5h after injection in the bone marrow and a slight elution of 18F-FDG. CONCLUSION: The activity of the obtained 18F-FDG-labelled HSPCs was sufficient to perform the micro-PET-CT imaging. Although the radiolabelling had a significant toxicity on HSPCs 15h after labelling, this technique allowed monitoring the beginning of HSPC homing into the bone marrow.
Authors: Mark G MacAskill; Adriana S Tavares; Junxi Wu; Christophe Lucatelli; Joanne C Mountford; Andrew H Baker; David E Newby; Patrick W F Hadoke Journal: Sci Rep Date: 2017-03-13 Impact factor: 4.379
Authors: Fernando A Oliveira; Mariana P Nucci; Igor S Filgueiras; João M Ferreira; Leopoldo P Nucci; Javier B Mamani; Fernando Alvieri; Lucas E B Souza; Gabriel N A Rego; Andrea T Kondo; Nelson Hamerschlak; Lionel F Gamarra Journal: Cells Date: 2020-04-10 Impact factor: 6.600
Authors: Fernando A Oliveira; Mariana P Nucci; Javier B Mamani; Arielly H Alves; Gabriel N A Rego; Andrea T Kondo; Nelson Hamerschlak; Mara S Junqueira; Lucas E B de Souza; Lionel F Gamarra Journal: Biomedicines Date: 2021-06-29