Microfluidic device for cell separation and uses thereof

• US 20070172903A1
• Filed: 03/21/2007
• Published: 07/26/2007
• 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 8,304,230 B2
• Time in Patent Office: Days
• US Class Current: 435/7.230
• 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 2015/1081 : Sorting the 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 device comprising a first array of obstacles disposed to selectively enrich a first cell type in a sample by a factor of at least 100,000.

2. The device of claim 1, wherein said first cell type is at a concentration of up to 50 cells/mL.

3. The device of claim 1, wherein said device retains at least 60% of said first cell type present in said sample.

12. The device of claim 1, 4, or 5, wherein said first array of obstacles is coupled to a substrate or a flow through chamber.

13. The device of claim 12, wherein said substrate comprises a polymer.

14. The device of claim 1, 4, or 5, wherein said first array of obstacles is a two dimensional array of obstacles.

15. The device of claim 1, 4, or 5, wherein said device is reusable.

39. The device of claim 1, 4, 5, or 38, wherein said device does not damage cells flowing there through.

40. The device of claim 1, 4, 5, or 38, wherein said device is capable of processing 20 mL sample in less than 60 minutes.

41. The device of claim 1, 4, 5, or 38, wherein said first array of obstacles is non-uniform.

44. The device of claim 40, wherein at least 60% of fetal cells in said sample are retained.

4. A device comprising a first array of obstacles that depletes one or more non-tumor cells from a blood sample and retains at least 60% of said tumor cells from said blood sample.

5. A device for enrichment of rare cells comprising a first array of obstacles adapted to enrich rare cells from a fluid sample by a factor of at least 100,000 and retain at least 60% of said rare cells.

6. The device of claim 5, wherein said rare cells are tumor cells or epithelial cells.

7. The device of claim 5, wherein said rare cells are fetal cells.

8. The device of claim 7, wherein said fetal cell is selected from the group consisting of fetal nucleated red blood cells, trophoblasts, and fetal fibroblasts.

9. The device of claim 8, wherein said fetal cell is a nucleated fetal red blood cell.

10. The device of claim 5, wherein said fluid sample is a maternal red blood sample.

11. The device of claim 10, wherein at least 99% of fetal cells present in said maternal blood sample are retained.

16. A method for producing an enriched cell population comprising a first cell type from a sample comprising two or more cell types comprising:

flowing said sample through an array of obstacles that selectively enriches said first cell type by a factor of at least 100,000.

17. The method of claim 16, wherein said enriched cell population comprises at least 60% of all first cell types present in said sample.

18. The method of claim 16, wherein said first cell type is present in said sample at a concentration of up to 50 cells/μ

L.

19. The method of claim 16, wherein said first cell type is a fetal cell and said sample is a maternal blood sample.

20. The method of claim 19, wherein said fetal cell is selected from the group consisting of a fetal nucleated red blood cell, trophoblast, and fetal fibroblast

21. The method of claim 20, wherein said fetal cell is a nucleated fetal red blood cell.

22. The method of claim 16, wherein the first cell type is a maternal red blood cell.

23. The method of claim 19, wherein said enriched cell population comprises at least 99% of fetal cells present in said maternal blood sample.

24. The method of claim 16, wherein said first cell type is an epithelial or tumor cell and said sample is a blood sample.

25. A method diagnosing or monitoring cancer in a patient comprising:

flowing a sample from said patient through a device comprising a plurality of obstacles on a substrate coupled to one or more binding moieties.

26. The method of claim 25, wherein said sample is flowed at a rate of at most 15 μ

l/sec.

27. The method of claim 25, wherein said binding moieties selectively bind to a rare cell.

28. The method of claim 27, wherein said rare cell is an epithelial cell or tumor cell.

29. The method of claim 25, wherein said sample is a blood sample.

30. The method of claim 25, wherein said device has sensitivity or specificity greater than 75%.

31. The method of claim 25, wherein said device retains at least 60% one or more rare cells from said sample.

32. The method of claim 25, wherein said device increases concentration of a rare cell from said sample by a factor of at least 100,000.

33. A device for cancer diagnosis or prognosis comprising an array of obstacles that selectively retain one or more rare cells from a sample wherein said array is configured to enable flow of cells without mechanically damaging them.

34. A method for diagnosing or prognosing cancer in a patient comprising:

flowing a sample from said patient through an array of obstacles without mechanically damaging rare cells in said sample.

36. A method for diagnosing or prognosing cancer comprising:

enriching one or more rare cells from a sample by flowing said sample through an array of obstacles;

performing nucleic acid analysis on said enriched rare cells.

35. The method of claim 36, wherein said sample flows at a rate of at most 15 μ

l/sec.

37. The method of claim 36 wherein said rare cells are epithelial cells or tumor cells.

38. A device comprising a plurality of obstacles that selectively retains one or more fetal cells from a sample comprising fetal cells and non-fetal cells.

42. The device of claim 38, wherein said device is a flow-through device.

43. The device of claim 38, wherein said device enriches said fetal cells by a factor of at least 100,000.