Methods and Apparatus for the Isolation and Enrichment of Circulating Tumor Cells

• US 20080248499A1
• Filed: 06/20/2007
• Published: 10/09/2008
• Est. Priority Date: 08/11/2005
• Status: Active Grant

• Current Assignee: University of Washington
• Original Assignee: University of Washington
• Inventors: Chiu, Daniel T., Kuo, Jason S.
• Granted Patent: US 7,993,821 B2
• Time in Patent Office: Days
• US Class Current: 435/7.23
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2400/0409 centrifugal forces
• CPC Class Codes: B01L 2400/0415 electrical forces, e.g. electrokinetic
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 3/502746 characterised by the means for controlling flow resistance, e.g. flow controllers, baffles B01L3/502738 takes precedence
• CPC Class Codes: B01L 3/502753 characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation separation in general B01D; microapparatus for analysis using electrophoresis G01N27/44791; sample preparation G01N1/28
• CPC Class Codes: C12N 5/0693 Tumour cells; Cancer cells
• CPC Class Codes: G01N 1/34 Purifying; Cleaning processes or apparatus for extracting or separating nucleic acids from biological samples C12N15/1003

Abstract

Embodiments in accordance with the present invention relate to methods and apparatuses for concentrating and isolating Circulating Tumor Cells (CTCs) from body fluids. One embodiment of the present invention includes a micro-fabricated or nano-fabricated device having channels configured for separating and excluding. Embodiments in accordance with the present invention utilize features that reduce the hydrodynamic pressure experienced by the cells during the separation, isolation and concentration processes, and therefore reduce the likelihood of cell lysis or other damage to the cells.

Claims

1. A method of isolating cancer cells, the method comprising:

passing a biofluid through a flow channel enclosed within walls and having a feature configured to impede movement of a cancer cell in a direction while allowing flow of the biofluid in the direction, wherein movement of the cancer cell is impeded by a cross-sectional shape of the flow channel and/or a relief on a surface of the flow channel wall.

2. The method of claim 1 further comprising diluting the biofluid prior to passage through the flow channel.

3. The method of claim 1 further comprising stabilizing the biofluid with a fixative prior to passage through the flow channel.

4. The method of claim 1 further comprising treating the biofluid with a material that binds preferentially to the cancer cell to assist in visualization of the cancer cells.

5. The method of claim 4 wherein the biofluid is treated with the material having fluorescent, magnetic, electroactive, bioactive, or photoactive properties.

6. The method of claim 1 further comprising visualizing the cancer cell with a microscope through a transparent wall of the channel.

8. The method of claim 1 wherein the direction is against a wall of a constriction in the channel, against a wall of a curvature in the channel, or against a wall of an expanded region of the channel.

9. The method of claim 1 further comprising:

collecting the cancer cell; and

treating the collected cancer cell with a material having fluorescent, magnetic, electroactive, bioactive, or photoactive properties.

10. The method of claim 9 wherein the material is configured to identify a subpopulation of cancer cells.

11. The method of claim 10 subpopulation of cancer cells comprises a putative cancer stem cell.

7. (canceled)

12-15. -15. (canceled)

16. A method comprising:

collecting one or more cancer cells by flowing a sample of a biofluid obtained from a patient from a first flow channel through an aperture defined by a smallest dimension of a second flow channel, smallest dimension smaller than a dimension of the one or more cancer cells, while allowing fluid to flow around the one or more cancer cells into the second flow channel; and

exposing the cancer cells to a material configured to bind preferentially thereto; and

counting a number of the collected cancer cells in order to evaluate a likelihood of metastasis.

17. The method of claim 16 wherein the cancer cells are exposed to the material prior to collection.

18. The method of claim 16 wherein the cancer cells are exposed to the material after the collection.

19. The method of claim 16 further comprising determining cancer diagnosis or prognosis based upon the number of cancer cells collected.

20. The method of claim 16 further comprising determining an effectiveness of cancer treatment based upon the number of cancer cells collected.