Microfluidic device for cell separation and uses thereof

• US 8,304,230 B2
• Filed: 03/21/2007
• Issued: 11/06/2012
• Est. Priority Date: 09/27/2002
• Status: Active Grant

• Current Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Original Assignee: The General Hospital Corporation (Mass General Brigham, Inc.)
• Inventors: Toner, Mehmet, Truskey, George, Kapur, Ravi
• Primary Examiner(s): Yu, Melanie J
• Application Number:
  US11/726,230
• Publication Number:
  US 20070172903A1
• Time in Patent Office: 2,057 Days
• Field of Search: 435/283.1, 435/287.1, 435/287.2, 435/288.7, 436/518, 422/50, 422/61, 422/68.1
• US Class Current: 435/288.5
• CPC Class Codes: B01L 1/52 Transportable laboratories; Field kits
• CPC Class Codes: B01L 2200/0647 Handling flowable solids, e.g. microscopic beads, cells, particles
• CPC Class Codes: B01L 2200/0652 Sorting or classification of particles or molecules
• CPC Class Codes: B01L 2200/0668 Trapping microscopic beads
• CPC Class Codes: B01L 2200/12 Specific details about manufacturing devices
• CPC Class Codes: B01L 2300/0681 Filter
• CPC Class Codes: B01L 2300/0816 Cards, e.g. flat sample carriers usually with flow in two horizontal directions
• CPC Class Codes: B01L 2300/0864 comprising only one inlet and multiple receiving wells, e.g. for separation, splitting
• CPC Class Codes: B01L 2300/0867 Multiple inlets and one sample wells, e.g. mixing, dilution
• CPC Class Codes: B01L 2300/0877 Flow chambers
• CPC Class Codes: B01L 2300/0883 Serpentine channels
• CPC Class Codes: B01L 2300/12 Specific details about materials
• CPC Class Codes: B01L 2400/0487 fluid pressure, pneumatics
• CPC Class Codes: B01L 2400/086 using baffles or other fixed flow obstructions
• 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: B01L 3/502761 specially adapted for handling suspended solids or molecules independently from the bulk fluid flow, e.g. for trapping or sorting beads, for physically stretching molecules investigating characteristics of particles G01N15/00
• CPC Class Codes: B01L 9/54 Supports specially adapted for pipettes and burettes automated pipetting stations G01N35/10
• CPC Class Codes: B33Y 80/00 Products made by additive manufacturing
• CPC Class Codes: B82Y 30/00 Nanotechnology for materials or surface science, e.g. nanocomposites
• CPC Class Codes: C12N 5/0087 : Purging against subsets of blood cells, e.g. purging alloreactive T cells
• CPC Class Codes: G01N 1/40 : Concentrating samples
• CPC Class Codes: G01N 1/405 : by adsorption or absorption
• CPC Class Codes: G01N 2015/1006 : for cytology
• CPC Class Codes: G01N 2015/1028 : Sorting particles
• CPC Class Codes: G01N 2333/70582 : CD71
• CPC Class Codes: G01N 2333/70589 : CD45
• CPC Class Codes: G01N 2333/70596 : Molecules with a "CD"-designation not provided for elsewhere in G01N2333/705
• CPC Class Codes: G01N 33/5091 : for testing the pathological state of an organism
• CPC Class Codes: G01N 33/54366 : Apparatus specially adapted for solid-phase testing
• CPC Class Codes: G01N 33/54386 : Analytical elements
• CPC Class Codes: G01N 33/56966 : Animal cells
• CPC Class Codes: G01N 33/574 : for cancer
• CPC Class Codes: Y10T 29/49982 : Coating
• CPC Class Codes: Y10T 428/24744 : Longitudinal or transverse tubular cavity or cell

Abstract

The invention features methods for separating cells from a sample (e.g., separating fetal red blood cells from maternal blood). The method begins with the introduction of a sample including cells into one or more microfluidic channels. In one embodiment, the device includes at least two processing steps. For example, a mixture of cells is introduced into a microfluidic channel that selectively allows the passage of a desired type of cell, and the population of cells enriched in the desired type is then introduced into a second microfluidic channel that allows the passage of the desired cell to produce a population of cells further enriched in the desired type. The selection of cells is based on a property of the cells in the mixture, for example, size, shape, deformability, surface characteristics (e.g., cell surface receptors or antigens and membrane permeability), or intracellular properties (e.g., expression of a particular enzyme).

Claims

1. A flow-through device for enriching tumor cells or epithelial cells, or both, in a blood sample, the device comprising

a flow chamber through which the blood sample flows along a flow path;

an array of obstacles distributed in the flow chamber, wherein the obstacles in the array are arranged in at least two rows aligned normal to the flow path, wherein the obstacles are equally spaced within each row in the array, and each successive row of obstacles is shifted in a direction normal to the flow path relative to the obstacles in the preceding row;

wherein (i) a distance between the obstacles varies in the flow direction and (ii) the obstacles have the same shape; and

wherein the array of obstacles forms a network of microfluidic channels disposed to selectively enrich tumor cells or epithelial cells, or both in a blood sample flowed through the device by a factor of at least 1000.

2. The device of claim 1, wherein the device selectively enriches tumor cells.

3. The device of claim 1, wherein the device retains at least 60% of the tumor cells or epithelial cells present in the blood sample.

4. The device of claim 1, wherein a row of obstacles in the array is shifted by a distance equal to a radius of capture of the tumor cells or epithelial cells.

5. The device of claim 1, wherein said device enriches tumor cells or epithelial cells, or both, from said blood sample from said patient by a factor of at least 100,000.

6. The device of claim 1, wherein the array is configured to enable flow of cells without mechanically damaging them.

7. The device of claim 1, wherein the obstacles in each of the rows in the array are spaced apart by at most 100 μ

m, and wherein the obstacles in a given row of obstacles in the array are shifted in a direction normal to the flow path by a distance of at most 50 μ

m-with respect to obstacles in an adjacent row.

8. The device of claim 1, wherein the array of obstacles is coated with a binding moiety that selectively binds to epithelial cells.

9. The device of claim 1, wherein the array of obstacles is coated with a binding moiety that selectively binds to tumor cells.

10. The device of claim 1, wherein a distance between edges of two adjacent obstacles in the array of obstacles is larger than a size of a largest cell in the blood sample.

11. The device of claim 1, wherein the obstacles are circular in cross-section.

12. The device of claim 1, wherein a diameter of an obstacle is 5 μ

m, 10 μ

m, 25 μ

m, 50 μ

m, 75 μ

m, 100 μ

m, or 250 μ

m.

13. The device of claim 1, wherein each obstacle is 5 μ

m, 10 μ

m, 25 μ

m, 50 μ

m, 75 μ

m, or 250 μ

m at a widest point of the obstacle.

14. The device of claim 1, wherein a spacing between the obstacles is 10 μ

m, 25 μ

m, 50 μ

m, 75 μ

m, 100 μ

m, 250 μ

m, 500 μ

m, or 750 μ

m.

15. The device of claim 8, wherein the binding moiety is an antibody.

16. The device of claim 15, wherein the binding moiety is an antibody to a cell surface receptor.

17. The device of claim 1, wherein the obstacles in the array are spaced at most 100 μ

m center to center.

18. The device of claim 1, wherein the obstacles in the array are spaced at least 10 μ

m apart.

19. The device of claim 1, wherein the obstacles in the array are spaced at most 500 μ

m apart.

20. The device of claim 1, wherein said device is composed of plastic.

21. The device of claim 1, wherein the array of obstacles is an ordered array of obstacles.

22. The device of claim 1, wherein the obstacles contact both the top and bottom of the chamber.

23. The device of claim 1, wherein there is a gap between the obstacles and one surface of the flow chamber.

24. The device of claim 1, coupled to a positive pressure pump.

25. The device of claim 1, coupled to a negative pressure pump.

26. The device of claim 1, coupled to an imaging system.

27. The device of claim 1, wherein a spacing between obstacles is at least 10 μ

m.

28. The device of claim 1, wherein a spacing between obstacles is at most 100 μ

m.

29. A device comprising

a flow chamber comprising an inlet and an outlet and a flow path from the inlet to the outlet in which a fluid sample may flow;

a substrate that forms a bottom of the flow chamber;

a plurality of obstacles arranged in a first array and a second array on the substrate,

wherein the first array comprises a plurality of obstacles arranged in two or more rows aligned normal to the flow path and forming a plurality of equilateral triangular arrays of obstacles, wherein each obstacle has an equal center-to-center distance to any adjacent obstacle,

wherein the second array comprises a plurality of obstacles arranged in two or more rows aligned normal to the flow path and forming a plurality of triangular arrays of obstacles, wherein one or more of the obstacles in the second array are shifted to have a first center-to-center distance to a first adjacent obstacle that is different than a second center-to-center distance to a second adjacent obstacle

and further wherein at least the first or second array of obstacles, or both, is coated with a binding moiety that selectively binds to a first type of cell in the sample, wherein the first type of cell is a tumor cell or epithelial cell.

30. The device of claim 29, wherein the obstacles shifted in the second array are shifted by a distance equal to a radius of capture of the first type of cell.

31. The device of claim 29, wherein the binding moiety is an antibody.

32. The device of claim 31, wherein the binding moiety is an antibody to a cell surface receptor.