Microfluidic device for cell separation and uses thereof

• US 20070259424A1
• Filed: 03/21/2007
• Published: 11/08/2007
• 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, Kapur, Ravi, Truskey, George
• Granted Patent: US 8,986,966 B2
• Time in Patent Office: Days
• US Class Current: 435/372
• 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 comprising:

i. providing a first substrate;

ii. creating a pattern of obstacles on said first substrate, and

iii. coating said pattern of obstacles with one or more binding moieties that selectively bind to one or more first cell types in a mixture of cells, wherein said first cell type is a fetal nucleated red blood cell.

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

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

5. The method of claim 3, wherein the metal-based substrate is a steel substrate or a nickel substrate.

6. The method of claim 3, wherein the polymeric-based substrate is a charged polymer or an uncharged polymer.

7. The method of claim 3, 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.

8. The method of claim 1 or 2, wherein the creating step comprises the use of lithography, micromachining, casting, molding, embossing, wet chemical etch, dry chemical etch, milling, diamond cutting, electroforming, LIGA, or any combination thereof.

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

10. The method of claim 8, wherein micromachining comprises laser micromachining, thin-film surface micromachining, silicon micromachining or plastic micromachining.

11. The method of either claim 8, wherein molding is injection molding.

12. The method of either claim 8, wherein the dry chemical etch is a deep reactive ion etch.

13. The method of claim 1 or 2, wherein the creating step further comprises bonding a second substrate to the first substrate.

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

15. The method of claim 13, wherein the second substrate material and the first substrate material are the same.

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

17. The method of claim 13, wherein the second substrate material and the substrate material are different.

18. The method of claim 13, 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 a different plastic.

19. The method of claim 1 or 2, wherein the height of each obstacle is about 100 μ

m.

20. The method of claim 1 or 2, wherein the shape of each obstacle is circular, semi-circular, oval, trapezoidal, rectangular, a square, or any combination thereof.

21. The method of claim 19, 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.

22. The method of claim 1 or 2, wherein the pattern of obstacles is a two dimensional array.

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

24. The method of claim 1 or 2, wherein the first substrate and the obstacles are treated with an oxygen plasma, a silane reagent, or a combination thereof.

25. The method of claim 1 or 2, 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.

26. The method of claim 1 or 2, 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.

27. The method of claim 1 or 2 wherein said binding moieties selectively bind trophoblasts.

31. The method of claim 1 or 2 further comprising the step of sonically drilling one or more inlets and one or more outlets.

2. A method for manufacturing a device comprising:

i. providing a first substrate;

ii. creating a pattern of obstacles on said first substrate wherein said pattern is non-uniform, and

iii. coating said pattern of obstacles with one or more binding moieties that selectively bind to one or more first cell types in a mixture of cells.

28. The method of claim 2 wherein said binding moieties selectively bind fetal nucleated red blood cells.

29. The method of claim 2 wherein said binding moieties selectively bind tumor cells.

30. The method of claim 2 wherein said binding moieties selectively bind epithelial cells.

32. The method of claim 2 wherein said coating step comprises coating a first binding moiety in a first region and a second binding moiety in a second region.

33. The method of claim 33 wherein said first region is in series with said second region.

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