Cellular labeling for nuclear magnetic resonance techniques

• US 8,147,806 B2
• Filed: 01/11/2005
• Issued: 04/03/2012
• Est. Priority Date: 01/16/2004
• Status: Active Grant

• Current Assignee: Carnegie Mellon University
• Original Assignee: Carnegie Mellon University
• Inventors: Ahrens, Eric T.
• Primary Examiner(s): Hartley, Michael G
• Assistant Examiner(s): SAMALA, JAGADISHWAR RAO
• Application Number:
  US10/586,015
• Publication Number:
  US 20080292554A1
• Time in Patent Office: 2,639 Days
• Field of Search: 424/9, 424/9.5, 424/9.6, 424/9.8, 424/450, 514/450, 514/832, 514/937
• US Class Current: 424/9.6
• CPC Class Codes: A61K 49/0002 General or multifunctional contrast agents, e.g. chelated agents
• CPC Class Codes: A61K 49/1806 Suspensions, emulsions, colloids, dispersions
• CPC Class Codes: A61K 51/04 Organic compounds
• CPC Class Codes: A61K 51/1251 micro- or nanospheres, micro- or nanobeads, micro- or nanocapsules

Abstract

The disclosure provides, in part, fluorocarbon imaging reagents and formulations for the ex vivo labeling of cells. Labeled cells may be detected in vivo or ex vivo by a nuclear magnetic resonance technique, such as magnetic resonance imaging (MRI) or magnetic resonance spectroscopy (MRS). The disclosure additionally provides methods for using the imaging reagents in a variety of clinical procedures.

Claims

1. A method for detecting a cell in a subject comprising:

a. administering to the subject a cell that is labeled with a fluorocarbon imaging reagent, wherein at least a portion of the fluorocarbon imaging reagent is internalized into the cell; and

b. examining at least a portion of the subject by ¹⁹F magnetic resonance imaging, thereby detecting the cell that is labeled with the fluorocarbon imaging reagent in the subject.

4. The method of claim 1, further comprising contacting the cell ex vivo with a fluorocarbon imaging reagent under conditions such that the fluorocarbon imaging reagent becomes associated with the cell prior to step a.

5. The method of claim 1, wherein the fluorocarbon imaging reagent is a perfluoropolyether.

6. The method of claim 1, wherein the cell is contacted with the fluorocarbon imaging reagent in the presence of an uptake enhancing reagent.

7. The method of claim 6, wherein the uptake enhancing reagent comprises a cationic lipid.

8. The method of claim 1, wherein at least a portion of the fluorocarbon imaging reagent is associated with the extracellular surface of the cell.

9. The method of claim 1, wherein the fluorocarbon imaging reagent is conjugated to a cellular targeting moiety.

10. The method of claim 9, wherein the cellular targeting moiety comprises an antibody that binds to an epitope that is exposed to the extracellular milieu.

11. The method of claim 1, wherein the fluorocarbon imaging reagent is conjugated to an internalization moiety.

12. The method of claim 1, wherein the cell is a mammalian cell.

13. The method of claim 1, wherein the cell is a cell of the immune system.

14. The method of claim 1, wherein the cell is a dendritic cell.

15. The method of claim 1, wherein the fluorocarbon imaging reagent is formulated as an emulsion.

16. The method of claim 1, wherein the emulsion comprises particles having a mean diameter of between 30 and 500 nm.

17. The method of claim 1, wherein the fluorocarbon imaging reagent is a perfluoro-crown ether.

18. The method of claim 17, wherein the imaging reagent is a perfluoro-15-crown-5-ether.

19. The method of claim 1, wherein the fluorocarbon is a perfluorinated polyether having an average formula:

XO(Y—

O)_nZ

wherein Y is selected from the group consisting of;

20. The method of claim 1, wherein the imaging reagent comprises an additional functional moiety.

21. The method of claim 20, wherein the additional functional moiety is a detection moiety.

22. The method of claim 21, wherein the detection moiety is selected from the group consisting of:

a fluorescent detection moiety and a PET detection moiety.

2. An ex vivo labeled cellular formulation comprising:

a. a cell; and

b. a fluorocarbon imaging reagent that is detectable by ¹⁹F magnetic resonance imaging and that is associated with the cell.

3. A method for detecting transplanted cells in a transplant recipient comprising:

a. administering cells for transplant to a transplant recipient, at least a portion of which cells for transplant are labeled with a fluorocarbon imaging reagent, wherein at least a portion of the fluorocarbon imaging reagent is internalized into the cell;

b. examining at least a portion of the subject by ¹⁹F magnetic resonance imaging, thereby detecting the cells that are labeled with the fluorocarbon imaging reagent in the subject.