Propagation of primary cells

• US 20080305473A1
• Filed: 03/13/2007
• Published: 12/11/2008
• Est. Priority Date: 03/13/2006
• Status: Abandoned Application

• Current Assignee: Veridex LLC (Johnson & Johnson)
• Original Assignee: Veridex LLC (Johnson & Johnson)
• Inventors: Mazumder, Abhijit, Chowdary, Dondapati, Burnett, Christine, Skelton, Joanne
• Application Number:
  US11/717,382
• Publication Number:
  US 20080305473A1
• Time in Patent Office: Days
• US Class Current: 435/6
• CPC Class Codes: A61P 35/00 Antineoplastic agents
• CPC Class Codes: C12Q 1/6886 for cancer immunoassay for cancer G01N33/574
• CPC Class Codes: C12Q 2600/118 Prognosis of disease development
• CPC Class Codes: C12Q 2600/154 Methylation markers
• CPC Class Codes: C12Q 2600/16 Primer sets for multiplex assays
• CPC Class Codes: C12Q 2600/178 miRNA, siRNA or ncRNA

Abstract

The present invention provides a method of propagating cells of interest obtained from a biological specimen by enriching the cells under conditions that maintain sufficient cell viability; and propagating the cells under conditions effective to allow cell viability, proliferation and integrity.

Claims

1. A method of propagating cells of interest obtained from a biological specimen comprising the steps of:

a) enriching the cells under conditions that maintain sufficient cell viability; and

b) propagating the cells under conditions effective to allow cell viability, proliferation and integrity.

2. The method according to claim 1 wherein the biological specimen is selected from urine, blood, serum, plasma, lymph, sputum, semen, saliva, tears, pleural fluid, pulmonary fluid, bronchial lavage, synovial fluid, peritoneal fluid, ascites, amniotic fluid, bone marrow, bone marrow aspirate, cerebrospinal fluid, tissue lysate or homogenate or a cell pellet.

3. The method according to claim 1 wherein the cells are used to determine the presence or absence of an indication.

4. The method according to claim 3 wherein the indication is cancer, risk assessment of inherited genetic pre-disposition, identification of tissue of origin of a cancer cell such as a CTC, identifying mutations in hereditary diseases, disease status (staging), prognosis, diagnosis, monitoring, response to treatment, choice of treatment (pharmacologic), infection (viral, bacterial, mycoplasmal, fungal), chemosensitivity drug sensitivity, metastatic potential or identifying mutations in hereditary diseases.

5. The method according to claim 1 wherein the cells are enriched by antibody/magnetic separation, fluorescence activated cell sorting, (FACs), filtration or manually.

6. The method according to claim 5 wherein the manual enrichment is by prostate massage.

7. The method according to claim 1 wherein the propagating is by culturing in vitro, ex vivo or in vivo.

8. The method according to claim 1 wherein proliferation is at least one cell doubling.

9. The method according to claim 1 wherein the integrity is determined by proliferation of cells of interest versus contaminating cells.

14. A composition comprising the cells obtained by the method of claim 1.

15. The composition of claim 14 wherein the cells are a clonal population.

16. A kit comprising biomarker detection agents for performing the method according to claim 1.

17. An article comprising biomarker detection agents for performing the method according to claim 1.

18. A method of making a cell product comprising, propagating cells according to claim 1 and harvesting a product produced by the cells.

19. The method according to claim 18 wherein the product is a protein.

20. The method according to claim 19 wherein the protein is an antibody, a cytokine, a cell surface protein or a recombinant protein.

21. A method of screening a chemicals for pharmaceutic efficacy comprising the steps of propagating cells according to claim 1 and subjecting the cells to the chemical and measuring the response of the cells to the chemical.

22. A method of making a primary cell line comprising the steps of propagating cells according to claim 1 and continuing propagation until the cells form a cell line.

23. A method of making a clonal cell population comprising the steps of propagating cells according to claim 1 and continuing selecting for a clonal population and propagation until the cells form a clonal cell population.

10. A method of determining metastatic potential of a cell from a biological specimen comprising the steps of:

a) enriching the cells under conditions that maintain sufficient cell viability; and

b) propagating the cells under conditions effective to allow cell viability, proliferation and integrity.

c) isolating nucleic acid and/or protein from the cells; and

d) analyzing the nucleic acid and/or protein to determine the presence, expression level or status of a Biomarker specific for metastatic potential.

11. A method of identifying mutations in hereditary diseases from a cell from a biological specimen comprising the steps of:

a) enriching the cells under conditions that maintain sufficient cell viability; and

b) propagating the cells under conditions effective to allow cell viability, proliferation and integrity.

c) isolating nucleic acid and/or protein from the cells; and

d) analyzing the nucleic acid and/or protein to determine the presence, expression level or status of a Biomarker specific for a hereditary disease.

12. A method of preserving genetic material from a cell from a biological specimen comprising the steps of:

a) enriching the cells under conditions that maintain sufficient cell viability; and

b) propagating the cells under conditions effective to allow cell viability, proliferation and integrity.

b) isolating nucleic acid and/or protein from the cells; and

c) preserving the nucleic acid and/or protein.

13. A method of making a tumor cell vaccine comprising the steps of

a) enriching the cells under conditions that maintain sufficient cell viability;

and

b) propagating the cells under conditions effective to allow cell viability, proliferation and integrity.

c) isolating nucleic acid and/or protein from the cells; and

d) using the nucleic acid and/or protein to formulate the vaccine