Microfluidic device for cell separation and uses thereof
First Claim
1. A method for producing a population enriched in a first type of cell from a mixture of at least three types of cells, said method comprising the steps of:
- (a) providing a mixture of cells comprising at least first, second, and third types of cells;
(b) contacting said mixture with a first microfluidic channel that selectively allows the passage therethrough of said first type of cell to produce a first cell population enriched in said first type of cells relative to said second type of cell; and
(c) contacting said first cell population with a second microfluidic channel that selectively allows the passage therethrough of said first type of cell to produce a second cell population enriched in said first type of cell relative to said second and third types of cells, thereby producing said population enriched in said first type of cell.
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Abstract
The invention features methods for separating cells from a sample (e.g., separating fetal red blood cells from maternal 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).
278 Citations
69 Claims
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1. A method for producing a population enriched in a first type of cell from a mixture of at least three types of cells, said method comprising the steps of:
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(a) providing a mixture of cells comprising at least first, second, and third types of cells;
(b) contacting said mixture with a first microfluidic channel that selectively allows the passage therethrough of said first type of cell to produce a first cell population enriched in said first type of cells relative to said second type of cell; and
(c) contacting said first cell population with a second microfluidic channel that selectively allows the passage therethrough of said first type of cell to produce a second cell population enriched in said first type of cell relative to said second and third types of cells, thereby producing said population enriched in said first type of cell. - View Dependent Claims (2, 3, 4, 5)
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6. A microfluidic system for the separation of one or more desired cells from a sample, said system comprising:
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(a) a lysis device comprising;
(i) at least two input channels;
(ii) a reaction chamber; and
(iii) an outlet channel, wherein said input channels are connected to said outlet channel through said reaction chamber;
(b) a cell binding device comprising obstacles disposed in a microfluidic channel, wherein said obstacles preferentially bind a first type of cell, compared to a second type of cell; and
(c) an arraying device comprising a two dimensional array of locations for the containment of individual cells. - View Dependent Claims (10, 11, 12, 13, 14, 15)
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7. A microfluidic system for the separation of one or more desired cells from a sample, said system comprising:
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(a) a lysis device comprising;
(i) at least two input channels;
(ii) a reaction chamber; and
(iii) an outlet channel, wherein said input channels are connected to said outlet channel through said reaction chamber; and
(b) a cell binding device comprising obstacles disposed in a microfluidic channel, wherein said obstacles preferentially bind a first type of cell, compared to a second type of cell.
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8. A microfluidic system for the separation of a desired cell from a sample, said system comprising:
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(a) a lysis device comprising;
(i) at least two input channels;
(ii) a reaction chamber; and
(iii) an outlet channel, wherein said input channels are connected to said outlet channel through said reaction chamber; and
(b) an arraying device comprising a two dimensional array of locations for the containment of individual cells.
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9. A microfluidic system for the separation of one or more desired cells from a sample, said system comprising:
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(a) a cell binding device comprising obstacles disposed in a microfluidic channel, wherein said obstacles preferentially bind a first type of cell, compared to a second type of cell; and
(b) an arraying device comprising a two dimensional array of locations for the containment of individual cells.
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16. A method for separating one or more cells of a second type from a sample, said method comprising the steps of:
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(a) introducing into one or more microfluidic channels (i) a sample comprising cells of at least a first and second type and (ii) a solution that preferentially lyses cells of the first type, to cause greater lysis of cells of the first type compared to cells of the second type;
(b) contacting the product of step (a) with a microfluidic device comprising obstacles disposed in a microfluidic channel, wherein said obstacles preferentially bind said second type of cell;
(c) collecting cells bound to said obstacles, thereby producing a cell population enriched in said second type of cell;
(d) arraying said cell population enriched in said second type of cell;
(e) identifying one or more cells of said second type in said population enriched in said second type of cell; and
(f) collecting said one or more cells of said second type, thereby separating said one or more cells of said second type from said sample. - View Dependent Claims (23, 24, 25, 26, 27, 28)
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17. A method for separating one or more cells of a second type from a sample, said method comprising the steps of:
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(a) introducing into one or more microfluidic channels (i) a sample comprising cells of at least a first, second, and third type and (ii) a solution that preferentially lyses cells of the first type, to cause greater lysis of cells of the first type compared to cells of the second type;
(b) contacting the product of step (a) with a microfluidic device comprising obstacles disposed in a microfluidic channel, wherein said obstacles preferentially bind said third type of cell compared to said second type of cell;
(c) collecting cells not bound to said obstacles, thereby producing a cell population enriched in said second type of cell;
(d) arraying said cell population enriched in said second type of cell;
(e) identifying one or more cells of said second type in said cell population enriched in said second type of cell; and
(f) collecting said one or more cells of said second type, thereby separating said one or more cells of said second type from said sample.
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18. A method for producing a population of cells enriched in a second type of cell, said method comprising the steps of:
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(a) introducing into one or more microfluidic channels (i) a sample comprising cells of at least a first and second type and (ii) a solution that preferentially lyses cells of the first type, to cause greater lysis of cells of the first type compared to cells of the second type;
(b) contacting the product of step (a) with a microfluidic device comprising obstacles disposed in a microfluidic channel, wherein said obstacles preferentially bind said second type of cell; and
(c) collecting cells bound to said obstacles, thereby producing said population of cells enriched in said second type of cell. - View Dependent Claims (29)
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19. A method for producing a population of cells enriched in a second type of cell, said method comprising the steps of:
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(a) introducing into one or more microfluidic channels (i) a sample comprising cells of at least a first, second, and third type and (ii) a solution that preferentially lyses cells of the first type, to cause greater lysis of cells of the first type compared to cells of the second type;
(b) contacting the product of step (a) with a microfluidic device comprising obstacles disposed in a microfluidic channel, wherein said obstacles preferentially bind said third type of cell compared to said second type of cell; and
(c) collecting cells not bound to said obstacles, thereby producing said population of cells enriched in said second type of cell.
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20. A method for separating one or more cells of a second type from a sample, said method comprising the steps of:
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(a) introducing into one or more microfluidic channels (i) a sample comprising cells of at least a first and second type and (ii) a solution that preferentially lyses cells of the first type, to cause greater lysis of cells of the first type compared to cells of the second type;
(b) arraying the product of step (a);
(c) identifying said one or more cells of said second type; and
(d) collecting said one or more cells of said second type, thereby separating said one or more cells of said second type from said sample.
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21. A method for separating one or more cells of a second type from a sample, said method comprising the steps of:
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(a) contacting a sample comprising cells of at least a first and second type with a microfluidic device comprising obstacles disposed in a microfluidic channel, wherein said obstacles preferentially bind said first type of cell compared to said second type of cell;
(b) collecting cells not bound to said obstacles, thereby producing a depleted cell population;
(c) arraying said depleted cell population;
(d) identifying said one or more cells of said second type in said depleted population; and
(e) collecting said one or more cells of said second type, thereby separating said one or more cells of said second type from said sample.
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22. A method for separating one or more cells of a second type from a sample, said method comprising the steps of:
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(a) contacting a sample comprising cells of at least a first and second type with a microfluidic device comprising obstacles disposed in a microfluidic channel, wherein said obstacles preferentially bind said second type of cell compared to said first type of cell;
(b) collecting cells bound to said obstacles, thereby producing a depleted cell population;
(c) arraying said depleted cell population;
(d) identifying said one or more cells of said second type in said depleted population; and
(e) collecting said one or more cells of said second type, thereby separating said one or more cells of said second type from said sample.
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30. A method of producing a cell population enriched in a second cell type, said method comprising the steps of:
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(a) providing a sample comprising cells of at least a first and second type;
(b) introducing into one or more microfluidic channels (i) said sample; and
(ii) a lysis buffer that preferentially lyses cells of the first type, to cause greater lysis of cells of the first type compared to cells of the second type, to yield said enriched population. - View Dependent Claims (31, 32, 33, 34, 35, 36, 37)
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38. A kit comprising:
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(a) a microfluidic device comprising;
(i) at least three input channels;
(ii) a reaction chamber comprising a serpentine channel;
(iii) a dilution chamber comprising a serpentine channel; and
(iv) an outlet channel; and
(b) a lysis buffer that preferentially lyses one cell type over another. - View Dependent Claims (39, 40, 41, 42, 43)
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44. A method of producing a depleted cell population, said method comprising the steps of:
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(a) contacting a sample comprising cells of at least a first and second type with a microfluidic device comprising obstacles that preferentially bind said first type of cell compared to said second type of cell; and
(b) collecting cells bound to said obstacles or collecting cells not bound to said obstacles, thereby producing a depleted cell population. - View Dependent Claims (45, 46, 47, 48, 49, 50)
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- 51. A microfluidic device comprising obstacles disposed in a microfluidic channel, each of which is coated with a binding moiety.
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57. A microfluidic device comprising:
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(a) a first region of obstacles disposed in a microfluidic channel defining a fluid flow path, wherein said obstacles in said first region preferentially bind a first type of cell compared to a second type of cell; and
(b) a second region of obstacles disposed in said microfluidic channel, wherein said obstacles in said second region preferentially bind a third type of cell compared to a fourth type of cell, provided that said first and third types of cells are not the same. - View Dependent Claims (58, 59, 60, 61, 62)
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63. A microfluidic device comprising:
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(a) a first region of obstacles disposed in a microfluidic channel defining a fluid flow path, wherein said obstacles in said first region preferentially bind a first type of cell compared to a second type of cell, wherein said obstacles are arranged in at least two substantially parallel rows and in at least two substantially parallel columns; and
(b) a second region of obstacles disposed in said microfluidic channel, wherein said obstacles in said second region preferentially bind a third type of cell compared to a fourth type of cell, wherein said obstacles are arranged in at least two substantially parallel rows and in at least two substantially parallel columns, wherein the first and second regions are disposed adjacent one another in a microfluidic channel, and wherein the rows in the second region are displaced relative to the rows in the first region by a distance of less than the distance between the rows in the first region. - View Dependent Claims (66, 67)
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64. The microfluidic device of 63, wherein the obstacles in the first region are coated with an anti-CD71, an anti-CD36, an anti-GPA, or an anti-CD45 antibody, or a combination thereof.
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65. The microfluidic device of 63, wherein the obstacles in the second region are coated with an anti-CD71, an anti-CD36, an anti-GPA, or an anti-CD45 antibody, or a combination thereof.
Specification