18F-FDG labelling of hematopoietic stem cells: Dynamic study of bone marrow homing by PET-CT imaging and impact on cell functionality

L Faivre; M Chaussard; L Vercellino; V Vanneaux; B Hosten; K Teixera; V Parietti; P Merlet; L Sarda-Mantel; N Rizzo-Padoin; J Larghero

doi:10.1016/j.retram.2016.06.002

¹⁸F-FDG labelling of hematopoietic stem cells: Dynamic study of bone marrow homing by PET-CT imaging and impact on cell functionality

Curr Res Transl Med. 2016 Jul-Sep;64(3):141-148. doi: 10.1016/j.retram.2016.06.002. Epub 2016 Sep 28.

Authors

L Faivre¹, M Chaussard², L Vercellino³, V Vanneaux⁴, B Hosten⁵, K Teixera⁶, V Parietti⁷, P Merlet⁸, L Sarda-Mantel⁹, N Rizzo-Padoin¹⁰, J Larghero¹¹

Affiliations

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.

PMID: 27765274
DOI: 10.1016/j.retram.2016.06.002

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.

Keywords: Fluorodeoxyglucose F18; Hematopoietic stem cells; Homing behavior; PET–CT; Radiolabelling; Radiotoxicity.

MeSH terms

Animals
Bone Marrow*
Cell Division
Cell Movement
Cells, Cultured
Clone Cells
Cord Blood Stem Cell Transplantation*
Fluorine Radioisotopes / analysis*
Fluorodeoxyglucose F18 / analysis*
Graft Survival
Hematopoietic Stem Cells / cytology*
Heterografts
Humans
Isotope Labeling / methods
Mice
Mice, Inbred NOD
Mice, SCID
Organ Specificity
Positron Emission Tomography Computed Tomography* / methods
Radiopharmaceuticals / analysis*
Viscera

Substances

Fluorine Radioisotopes
Radiopharmaceuticals
Fluorodeoxyglucose F18
Fluorine-18