Selective delivery of material to cells

Patent metadata

Field	Value
Publication number	US-10870112-B2
Application number	US-201816145865-A
Country	US
Kind code	B2
Filing date	Sep 28, 2018
Priority date	Aug 16, 2013
Publication date	Dec 22, 2020
Grant date	Dec 22, 2020

Abstract

Isolating or identifying a cell based on a physical property of said cell can include providing a cell suspension; passing said suspension through a microfluidic channel that includes a constriction; passing the cell suspension through the constriction; and, contacting said cell suspension solution with a compound. The constriction can be sized to preferentially deform a relatively larger cell compared to a relatively smaller cell.

First claim

The invention claimed is:

1. A method for delivering a compound into a cell based on a physical property of said cell comprising:
providing a cell suspension comprising a first group of cells and a second group of cells, wherein the first group of cells have a relatively different size, diameter, and/or membrane stiffness than the second group of cells;
passing said cell suspension through and out of a microfluidic channel that includes a constriction, said constriction being sized to deliver a compound to the first group of cells to a greater extent than the second group of cells, wherein passage through the constriction disrupts the cell membranes of the first group of cells such that the compound is delivered into the first group of cells to a greater extent than the second group of cells in the cell suspension that passes through the microfluidic channel; and
contacting said cell suspension with the compound before the cell suspension is passed through the microfluidic channel that includes the constriction.

2. The method of claim 1 , wherein the first group of cells have a relatively different size than the second group of cells.

3. The method of claim 1 , wherein the first group of cells have a relatively different diameter than the second group of cells.

4. The method of claim 1 , wherein the constriction has a diameter smaller than the size of cell in the cell suspension to which the compound is delivered.

5. The method of claim 1 , wherein the constriction has a diameter smaller than the size of the smallest cell of the first group of cells or the smallest cell of the second group of cells.

6. The method of claim 1 , wherein the constriction has a diameter smaller than the size of the largest cell of the first group of cells or the second group of cells.

7. The method of claim 1 , wherein the cell suspension is further contacted with the compound after the suspension is passed through the microfluidic channel comprising the constriction.

8. The method of claim 1 , wherein the cell suspension comprises whole blood.

9. The method of claim 1 , wherein the cell suspension comprises peripheral blood mononuclear cells (PBMCs).

10. The method of claim 1 , wherein the cell suspension comprises an erythrocycte-depleted population of peripheral blood cells.

11. The method of claim 1 , wherein the compound has a molecular mass of 0.5 kDa to 5 MDa or a molecular mass of 3 kDa to 10 kDa.

12. The method of claim 1 , wherein the compound comprises one or more of a protein, a nucleic acid, a detectable marker, an active biomolecule, and a toxin.

13. The method of claim 1 , wherein the compound comprises a detectable marker.

14. The method of claim 1 , wherein the constriction has a width from 4 μm-10 μm.

15. The method of claim 1 , wherein the speed of the cells traversing the constriction ranges from 10 mm/s to 10 m/s.

16. The method of claim 1 , further comprising applying a pressure to the cell suspension to drive the cell suspension through the constriction of the microfluidic channel.

17. The method of claim 1 , wherein the first group of cells and/or the second group of cells comprise a cell selected from the group consisting of: tumor cells, PBMCs, and erythrocytes.

18. The method of claim 17 , wherein the first group of cells are PBMCs.

19. The method of claim 17 , wherein the PBMCs comprise leukocytes.

20. The method of claim 19 , wherein the leukocytes comprise neutrophils, eosinophils, basophils, lymphocytes, and/or monocytes.

21. The method of claim 19 , wherein the leukocytes are lymphocytes.

22. The method of claim 1 , wherein the constriction has a length of 1 μm-100 μm.

23. The method of claim 1 , wherein the constriction has 1-10 constrictions in series.

24. The method of claim 1 , wherein the compound comprises a magnetic particle, a fluorescent particle, a fluorescent dye, a fluorescent protein, nucleic acid encoding a fluorescent protein, luciferase, or a compound that induces cell death.

25. The method of claim 1 , wherein the compound comprises a detectable marker.

26. The method of claim 25 , further comprising sorting the cells in the cell suspension based on uptake of the detectable marker, thereby isolating the first group of cells.

27. The method of claim 25 , further comprising tagging the cells in the cell suspension with an antibody and sorting the cells in the cell suspension based on uptake of the detectable marker and/or antibody tagging, thereby isolating the first group of cells.

28. The method of claim 27 , wherein the antibody binds a target on the first group of cells, and the cells are sorted by presence of the detectable marker and presence of the antibody tag, and wherein the antibody is a blood cell selective antibody.

29. The method of claim 27 , wherein the antibody binds a target on the second group of cells, and cells are sorted by presence of the detectable marker and absence of the antibody tag, and wherein the antibody is a blood cell selective antibody.

30. The method of claim 1 , wherein at least 10% more of the compound is delivered into the first group of cells compared to the second group of cells.

31. The method of claim 1 , wherein the first group of cells have a relatively different membrane stiffness than the second group of cells.

32. The method of claim 1 , wherein:
the first group of cells and/or the second group of cells comprise a cell selected from the group consisting of: tumor cells, PBMCs, and erythrocytes;
the first group of cells have a relatively different diameter than the second group of cells;
the constriction has a diameter smaller than the diameter of a cell of the first group of cells in the cell suspension to which the compound is delivered;
the constriction has a width from 4 μm-10 μm, a length of 1 μm-100 μm, and 1-10 constrictions in series; and
the compound has a molecular mass of 0.5 kDa to 5 MDa.

Assignees

• Massachusetts Inst Technology

Inventors

• Sharei Armon R
• Adalsteinsson Viktor A
• Cho Nahyun
• Langer Robert S
• Love J Christopher
• Jensen Klavs F

Classifications

• G01N33/575

  for cancer · CPC title

• C12N15/87

  Introduction of foreign genetic material using processes not otherwise provided for, e.g. co-transformation · CPC title

• B82Y30/00

  Nanotechnology for materials or surface science, e.g. nanocomposites · CPC title

• C12N5/0634

  Cells from the blood or the immune system · CPC title

• G01N2015/1006

  for cytology · CPC title