Methods for assessing cell labeling

• US 20070258886A1
• Filed: 04/16/2007
• Published: 11/08/2007
• Est. Priority Date: 04/14/2006
• Status: Abandoned Application

• Current Assignee: Celsense, Inc.
• Original Assignee: Celsense, Inc.
• Inventors: Ahrens, Eric, Kornblith, Paul
• Application Number:
  US11/787,520
• Publication Number:
  US 20070258886A1
• Time in Patent Office: Days
• US Class Current: 424/1.110
• CPC Class Codes: A61B 5/055 involving electronic [EMR] or nuclear [NMR] magnetic resonance, e.g. magnetic resonance imaging
• CPC Class Codes: A61B 6/037 Emission tomography
• CPC Class Codes: A61K 49/0002 General or multifunctional contrast agents, e.g. chelated agents
• CPC Class Codes: A61K 49/0021 the fluorescent group being a small organic molecule
• CPC Class Codes: A61K 49/0082 micelle, e.g. phospholipidic micelle and polymeric micelle
• CPC Class Codes: A61K 49/0093 Nanoparticle, nanocapsule, nanobubble, nanosphere, nanobead, i.e. having a size or diameter smaller than 1 micrometer, e.g. polymeric nanoparticle
• CPC Class Codes: A61K 49/0097 Cells, viruses, ghosts, red blood cells, viral vectors, used for imaging or diagnosis in vivo
• CPC Class Codes: A61K 49/1809 Micelles, e.g. phospholipidic or polymeric micelles
• CPC Class Codes: A61K 49/1896 not provided for elsewhere, e.g. cells, viruses, ghosts, red blood cells, virus capsides
• CPC Class Codes: G01R 33/5601 involving use of a contrast agent for contrast manipulation, e.g. a paramagnetic, super-paramagnetic, ferromagnetic or hyperpolarised contrast agent

Abstract

The disclosure relates to methods for labeling cells ex vivo with agents that can be detected by MRI or PET techniques, or other techniques that permit non-invasive imaging of living organisms. The labeled cells can be re-administered to the patient and the movements of the labeled cells can be tracked in vivo by MRI, PET or other techniques. In part, the disclosed methods involve labeling a series of cell samples ex vivo determining the association of label with the cells, such that an appropriate dosage of labeled cells can be determined for each patient.

Claims

1. A method for assessing the labeling of cells intended for administration to a patient, the method comprising:

a) contacting cells of a plurality of substantially identical cell samples with an in vivo imaging reagent;

b) detecting the in vivo imaging reagent in the samples; and

c) assessing the labeling of the cells with the in vivo imaging reagent.

3. The method of claim 1, wherein at least two of the substantially identical cell samples are subjected to predetermined conditions that differ from each other, and wherein the method further comprises assessing the effects of differing conditions on the labeling of the cells with the in vivo imaging reagent.

4. The method of claim 3, wherein at least ten substantially identical cell samples are subjected to predetermined conditions that differ from each other.

5. The method of claim 4, wherein the cells are contacted with the in vivo imaging reagent or the in vivo imaging reagent and the proxy reagent by use of automated, high-throughput technology.

6. The method of claim 5, wherein automated, high-throughput technology is used to assess the labeling of the cells with the in vivo imaging reagent.

7. The method of claim 1, wherein the in vivo imaging reagent is an agent that is suitable for detection in vivo by a nuclear magnetic resonance technique.

8. The method of claim 7, wherein in vivo imaging reagent is selected from the group consisting of:

paramagnetic imaging agents, superparamagnetic iron-oxide particles, magnetite particles, fluorocarbon imaging reagents, Gd chelates, and Mn chelates.

9. The method of claim 8, wherein the in vivo imaging reagent comprises PFPE emulsions.

10. The method of claim 1, wherein the in vivo imaging reagent is an agent that is suitable for detection by a non-invasive imaging technique selected from the group consisting of:

positron emission tomography (PET), gamma ray detection, or SPECT (single-photon emission computed tomography).

15. The method of claims 3, wherein the predetermined conditions that differ from each other differ in one or more aspect selected from the group consisting of:

temperature, atmospheric composition, humidity, culture medium concentration of the in vivo labeling agent, duration of exposure to the in vivo labeling agent, concentration of an uptake enhancing agent and duration of exposure to an uptake enhancing agent.

16. The method of claim 1, further comprising determining conditions for labeling the cells with the in vivo imaging reagent that will allow the preparation of a dosage of the cells for administration to a patient, wherein the dosage conforms to known safety parameters associated with the in vivo imaging reagent and provides adequate labeling to permit detection of the labeled cells in vivo.

17. The method of claim 1, further comprising administering to the patient a dosage of the cells, wherein the dosage conforms to known safety parameters associated with the in vivo imaging reagent and provides adequate labeling to permit detection of the labeled cells in vivo.

18. The method of claim 3, further comprising an additional assay selected from the group consisting of:

a viability assay, a cell count, a cell cycle assay, a migration assay, and a functional assay.

19. The method of claim 1, wherein the patient is a mammal.

20. The method of claim 1, wherein the patient is a human.

21. The method of claim 1, wherein the cells are autologous to the patient.

22. The method of claim 1, wherein the cells are allogeneic to the patient.

23. The method of claim 1, wherein cells are selected from the group comprising:

blood cells, myoblasts, bone marrow cells, peripheral blood cells, umbilical cord blood cells, cardiomyocytes, chondrocytes, immune cells, fetal neural cells, neuronal precursors, fibroblasts, hepatocytes, islet cells of pancreas, keratinocytes and precursors of any of the preceding.

24. The method of claim 1, wherein the cells are selected from the group consisting of:

embryonic stem cells, cells cultured from embryonic stem cells, adult stem cells and cells cultured from adult stem cells.

25. The method of claim 1, wherein the substantially identical samples are each a portion of a sample of cells obtained from a donor.

26. The method of claim 1, wherein the substantially identical samples are each a portion of cells cultured from cells derived from a donor.

29. The method of claim 1, wherein assessing the labeling of the cells with the in vivo imaging reagent comprises quantifying the amount of in vivo imaging reagent that is associated with the cells.

2. A method for assessing the labeling of cells intended for administration to a patient, the method comprising:

a) contacting cells of a plurality of substantially identical cell samples with an in vivo imaging reagent and a proxy reagent;

b) detecting the proxy reagent in the samples; and

c) assessing the labeling of the cells with the proxy reagent, thereby assessing the labeling of the cells with the in vivo imaging reagent.

11. The method of claim 2, wherein the proxy reagent is selected from the group consisting of:

a fluorescent or luminescent protein, a fluorescent or luminescent analogue agent, a fluorescent or luminescent dye, a colorimetric agent, or a radioactive agent.

12. The method of claim 2, wherein the in vivo imaging reagent is stably associated with the proxy reagent to form a dual imaging reagent.

13. The method of claim 12, wherein the in vivo imaging reagent is an agent that is suitable for detection in vivo by a nuclear magnetic resonance technique.

14. The method of claim 12, wherein the proxy reagent is selected from the group consisting of:

a fluorescent or luminescent protein, a fluorescent or luminescent analogue agent, a fluorescent or luminescent dye, a colorimetric agent, or a radioactive agent.

30. The method of claim 2, wherein assessing the labeling of the cells with the in vivo imaging reagent comprises quantifying the amount of proxy reagent that is associated with the cells.

27. A composition for labeling cells, the composition comprising an in vivo imaging reagent in stable association with a protein proxy reagent.

28. The composition of claim 27, wherein the protein proxy reagent is selected from the group consisting of:

a fluorescent protein, a luminescent protein, a colored protein, a fluorogenic protein, a luminogenic protein and a chromogenic protein.

31. A method of administering a safe and useful dosage of labeled cells to a patient and detecting said labeled cells in vivo, the method comprising:

a) contacting cells of a plurality of substantially identical cell samples with an in vivo imaging reagent;

b) detecting the in vivo imaging reagent in the samples;

c) assessing the labeling of the cells with the in vivo imaging reagent;

d) determining conditions for labeling the cells with the in vivo imaging reagent that will allow the preparation of a dosage of the cells for administration to a patient, wherein the dosage conforms to known safety parameters associated with the in vivo labeling agent and provides adequate labeling to permit detection of the labeled cells in vivo;

e) administering the dosage of the cells to the patient; and

f) detecting the labeled cells in vivo by a non-invasive imaging technique.

32. The method of claim 31, wherein the non-invasive imaging technique in a nuclear magnetic resonance technique.

33. A method of administering a safe and useful dosage of labeled cells to a patient and detecting said labeled cells in vivo, the method comprising:

a) contacting cells of a plurality of substantially identical cell samples with an in vivo imaging reagent and a proxy reagent;

b) detecting the proxy reagent in the samples;

c) assessing the labeling of the cells with the proxy reagent, thereby assessing the labeling of the cells with the in vivo imaging reagent;

d) determining conditions for labeling the cells with the in vivo imaging reagent that will allow the preparation of a dosage of the cells for administration to a patient, wherein the dosage conforms to known safety parameters associated with the in vivo labeling agent and provides adequate labeling to permit detection of the labeled cells in vivo;

e) administering the dosage of the cells to the patient; and

f) detecting the labeled cells in vivo by a non-invasive imaging technique.

34. The method of claim 33, wherein the non-invasive imaging technique in a nuclear magnetic resonance technique.