Cell division marker

• US 9,222,939 B2
• Filed: 04/12/2013
• Issued: 12/29/2015
• Est. Priority Date: 10/06/2005
• Status: Active Grant

• Current Assignee: University Of Massachusetts
• Original Assignee: University Of Massachusetts
• Inventors: Doxsey, Stephen J., Chen, Chun-Ting
• Primary Examiner(s): Gabel, Gail R
• Application Number:
  US13/862,143
• Publication Number:
  US 20130337472A1
• Time in Patent Office: 991 Days
• Field of Search: 435/2, 435/6, 435/7.1, 435/7.2, 435/7.21, 435/7.23, 435/7.24, 435 792- 795, 435/40.51, 435/347, 435/354, 435/358, 435/363, 435/366, 435/371, 435/372, 435/373, 435/374, 435/376, 435/976, 436/512, 436/517, 436/519, 436/63, 436/64, 436/164, 436/172, 436/813
• US Class Current: 1/1
• CPC Class Codes: C07K 14/705 Receptors; Cell surface antigens; Cell surface determinants tumour specific antigens C07K14/4748
• CPC Class Codes: G01N 2800/085 Liver diseases, e.g. portal hypertension, fibrosis, cirrhosis, bilirubin
• CPC Class Codes: G01N 2800/12 Pulmonary diseases
• CPC Class Codes: G01N 2800/20 Dermatological disorders
• CPC Class Codes: G01N 2800/323 Arteriosclerosis, Stenosis
• CPC Class Codes: G01N 2800/328 Vasculitis, i.e. inflammation of blood vessels
• CPC Class Codes: G01N 2800/342 Prostate diseases, e.g. BPH, prostatitis
• CPC Class Codes: G01N 2800/347 Renal failures; Glomerular diseases; Tubulointerstitial diseases, e.g. nephritic syndrome, glomerulonephritis; Renovascular diseases, e.g. renal artery occlusion, nephropathy
• CPC Class Codes: G01N 2800/36 Gynecology or obstetrics
• CPC Class Codes: G01N 2800/364 Endometriosis, i.e. non-malignant disorder in which functioning endometrial tissue is present outside the uterine cavity
• CPC Class Codes: G01N 33/564 for pre-existing immune complex or autoimmune disease , i.e. systemic lupus erythematosus, rheumatoid arthritis, multiple sclerosis, rheumatoid factors or complement components C1-C9
• CPC Class Codes: G01N 33/56966 Animal cells
• CPC Class Codes: G01N 33/57484 involving compounds serving as markers for tumor, cancer, neoplasia, e.g. cellular determinants, receptors, heat shock/stress proteins, A-protein, oligosaccharides, metabolites
• CPC Class Codes: G01N 33/57492 involving compounds localized on the membrane of tumor or cancer cells

Abstract

This application relates to a newly identified animal cell structure, the midbody scar. This structure is a remnant of the midbody that is retained by one daughter cell following cytokinesis and persists through multiple subsequent cell cycles. The midbody scar can be useful as a marker of dividing cells or of a cell'"'"'s replicative age.

Claims

1. A method of identifying a cancer stem cell in a sample comprising a population of mammalian cells, the method comprising:

providing a sample comprising a population of mammalian cells that is suspected of containing a cancer stem cell;

detecting a cell having two or more midbody scars by a method comprising contacting the sample with an antibody or an antigen-binding portion thereof that binds specifically to a polypeptide component of a midbody scar selected from the group consisting of;

mitotic kinesin-like protein 1 (MKLP1), Epsilon tubulin (ε

-tubulin), Centrosomal Protein 55KDa (Cep55), and Aurora B kinase;

detecting binding of the antibody or antigen-binding portion thereof to the polypeptide component, wherein a ring structure not at a midbody or centrosome is a midbody scar; and

identifying a cell in the sample having two or more midbody scars as being a cancer stem cell.

2. The method of claim 1, wherein a cell in the sample having more than four midbody scars is identified as a cancer stem cell.

3. The method of claim 1, wherein the sample is from a subject suspected of having a cancer.

4. The method of claim 3, wherein the subject is human.

5. The method of claim 4, wherein the sample comprises a population of cells from skin, heart, neural tissue, bone marrow, blood, liver, or lung.

6. The method of claim 5, wherein the sample comprises a population of cells from blood.

7. The method of claim 4, wherein the sample is a pathology specimen.

8. The method of claim 7, wherein the pathology specimen is a fixed tissue sample.

9. The method of claim 1 wherein the antibody or antigen-binding portion thereof is conjugated to a detectable label.

10. The method of claim 9, wherein the detectable label is selected from the group consisting of:

a fluorescent moiety, a radioactive isotope, an enzyme, a contrast agent, and a magnetic particle.

11. The method of claim 10, wherein the detectable label is a fluorescent moiety.

12. The method of claim 11, wherein the detecting of the binding of the antibody or antigen-binding portion thereof to the polypeptide component is performed by measuring the fluorescence of the cell contacted with the antibody or antigen-binding portion thereof.

13. The method of claim 12, wherein the detecting is automated.

14. The method of claim 13, wherein the automated detection is performed using fluorescence-assisted cell sorting.

15. The method of claim 10, wherein the detectable label is a magnetic particle.

16. The method of claim 15, wherein the detecting is automated.

17. The method of claim 16, wherein the automated detection is performed using magnetic-activated cell sorting.

18. The method of claim 1, further comprising isolating the identified cancer stem cell from the sample.

19. The method of claim 18, further comprising culturing the isolated cancer stem cell.