Microfluidic device for cell separation and uses thereof

• US 8,986,966 B2
• Filed: 03/21/2007
• Issued: 03/24/2015
• 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): Ware, Debbie K
• Application Number:
  US11/726,231
• Publication Number:
  US 20070259424A1
• Time in Patent Office: 2,925 Days
• Field of Search: None
• US Class Current: 435/174
• 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 method for manufacturing a device, the method comprising:

providing a first substrate;

forming a non-uniform array of obstacles on the first substrate, wherein the obstacles have a height between 25 μ

m and 150 μ

m, inclusive, and

coating the non-uniform array of obstacles with one or more binding moieties that selectively bind to one or more first cell types in a mixture of cells.

2. The method of claim 1, wherein the substrate is selected from a silicon-based substrate, a metal-based substrate, or a polymeric-based substrate.

3. The method of claim 2, wherein the silicon-based substrate is a glass substrate, a silicon substrate, or a silicon-on insulator substrate.

4. The method of claim 2, wherein the metal-based substrate is a steel substrate or a nickel substrate.

5. The method of claim 2, wherein the polymeric-based substrate is a charged polymer or an uncharged polymer.

6. The method of claim 2, wherein the polymeric-based substrate is a urethane polymer, an epoxy polymer, an acrylic polymer, or a polymer selected from the group consisting of poly(methylmethacrylate), polycarbonate, polystyrene, polyethylene, poly(dimethylsiloxane) and polyolefins.

7. The method of claim 1, wherein forming the non-uniform array of obstacles on said first substrate comprises forming by lithography, micromachining, casting, molding, embossing, wet chemical etch, dry chemical etch, milling, diamond cutting, electroforming, Lithographie Galvanoformung and Abformung (LIGA), or any combination thereof.

8. The method of claim 7, wherein lithography comprises photolithography, soft lithography, stereolithography or x-ray lithography.

9. The method of claim 7, wherein micromachining comprises laser micromachining, thin-film surface micromachining, silicon micromachining or plastic micromachining.

10. The method of claim 7, wherein molding is injection molding.

11. The method of claim 7, wherein the dry chemical etch is a deep reactive ion etch.

12. The method of claim 1, wherein forming the non-uniform array of obstacles on said first substrate further comprises bonding a second substrate to the first substrate.

13. The method of claim 12, wherein the bonding comprises the use of clamping, gluing, heating, anodic bonding, wafer bonding, a vacuum, or any combination thereof.

14. The method of claim 12, wherein the second substrate material and the first substrate material are the same.

15. The method of claim 14, wherein the first substrate and the second substrate are made of a polymeric material.

16. The method of claim 12, wherein the second substrate material and the first substrate material are different.

17. The method of claim 12, wherein the first substrate is glass and the second substrate is silicon, the first substrate is silicon and the second substrate is glass, the first substrate is glass and the second substrate is plastic, the first substrate is plastic and the second substrate is glass, or the first substrate is plastic and the second substrate is a different plastic.

18. The method of claim 1, wherein the height of each obstacle is about 100 μ

m.

19. The method of claim 1, wherein the shape of each obstacle is circular, semi-circular, oval, trapezoidal, rectangular, a square, or any combination thereof.

20. The method of claim 18, wherein the width, length, or diameter of the obstacle is at least 5 μ

m, at least 10 μ

m, at least 25 μ

m, at least 50 μ

m, at least 75 μ

m, at least, 100 μ

m, at least 250 μ

m, or at least 500 μ

m.

21. The method of claim 1, wherein the non-uniform array of obstacles is a two dimensional array.

22. The method of claim 21, wherein the two dimensional array comprises a plurality of rows wherein every subsequent row is shifted relative to a previous row.

23. The method of claim 1, further comprising treating the first substrate and the obstacles with an oxygen plasma, a silane reagent, or a combination thereof.

24. The method of claim 1, wherein the plurality of binding moieties are immobilized on the surface of the obstacles, physio-absorbed onto the surface of the obstacles, self-assembled on the surface of the obstacles, or any combination thereof.

25. The method of claim 1, wherein the binding moieties comprise an antibody, a charged polymer, a molecule that binds to a cell surface receptor, a polypeptide, a viral protein, a bacterial protein, a nucleic acid, or a carbohydrate.

26. The method of claim 1, wherein said binding moieties selectively bind trophoblasts.

27. The method of claim 1 wherein said binding moieties selectively bind fetal nucleated red blood cells.

28. The method of claim 1, wherein said binding moieties selectively bind tumor cells.

29. The method of claim 1, wherein said binding moieties selectively bind epithelial cells.

30. The method of claim 1, further comprising sonically drilling one or more inlets and one or more outlets on said first substrate.

31. The method of claim 1, wherein said coating step comprises coating a first binding moiety in a first region and a second binding moiety in a second region.

32. The method of claim 31, wherein said first region is in series with said second region.

33. The method of claim 31, wherein said first region is in parallel with said second region.

34. The method of claim 1, wherein the obstacles have a height between 50 μ

m and 100 μ

m, inclusive.

35. The method of claim 1, further comprising forming walls on the substrate to define a chamber of the device and providing a surface opposite the substrate and contacting the walls, wherein the height of the obstacles is selected to provide a gap between ends of the obstacles and the surface of the device.

36. The method of claim 1, wherein distances between obstacles are selected to be larger than the largest cells in said mixture of cells.

37. The method of claim 1, wherein the obstacles are arranged to enable cells to flow between the obstacles without being mechanically squeezed.

38. The method of claim 1, wherein spacing between the obstacles is between 10 μ

m and 250 μ

m.

39. A method for manufacturing a device comprising a chamber having a top surface and bottom surface, the method comprising:

providing a first substrate to form the top surface or bottom surface;

forming an array of obstacles on the first substrate, wherein the obstacles have a height less than a height of the chamber, and

coating a surface of the device with a binding moiety that selectively binds to one or more first cell types in a mixture of cells.

40. The method of claim 39, wherein said binding moiety comprises an antibody for a cell surface receptor.

41. The method of claim 39, wherein the obstacles have a height between 25 μ

m and 150 μ

m, inclusive.

42. The method of claim 41, wherein the obstacles have a height between 50 μ

m and 100 μ

m, inclusive.

43. The method of claim 41, wherein the height of each obstacle is about 100 μ

m.

44. The method of claim 39, wherein the obstacles are randomly distributed in the array.

45. The method of claim 39, wherein distances between obstacles are selected to be larger than the largest cells in said mixture of cells.

46. The method of claim 39, wherein the obstacles are arranged to enable cells to flow between the obstacles without being mechanically squeezed.

47. The method of claim 39, wherein spacing between the obstacles is between 10 μ

m and 250 μ

m.