Delivery of materials to anucleate cells

Patent metadata

Field	Value
Publication number	US-2018201889-A1
Application number	US-201815865901-A
Country	US
Kind code	A1
Filing date	Jan 9, 2018
Priority date	Jul 9, 2015
Publication date	Jul 19, 2018
Grant date	—

Abstract

The current subject matter includes methods, systems, articles, and techniques to deliver material to anucleate cells, such as red blood cells. Using a rapid deformation based microfluidic system, loading of red blood cells with macromolecules of different sizes has been shown. Although delivery to some mammalian cells, such as cancer cell lines and fibroblasts had been previously demonstrated using this technique, those designs were incompatible with RBCs that have dramatically different physical properties. Through the use of smaller constriction sizes, high speeds and different buffers successful delivery to red blood cells can be achieved. By enabling robust delivery to red blood cells in a simple, scalable manner, the current subject matter can be implemented in a diversity of applications that deliver material to study red blood cell diseases and/or use red blood cells as a therapeutic platform. Related apparatus, systems, techniques, and articles are also described.

First claim

1 . A microfluidic system for delivering a payload into an anucleate cell, the system comprising:
at least one microfluidic channel defining a lumen and being configured such that the anucleate cell suspended in a buffer can pass therethrough, wherein the microfluidic channel comprises at least one cell-deforming constriction comprising a length, a depth, and a width, wherein the width of the constriction is less than 4 micrometers.

2 - 26 . (canceled)

27 . A method for delivering a payload into an anucleate cell comprising:
passing the anucleate cell suspended in a buffer through a microfluidic channel that includes a cell-deforming constriction causing perturbations of the anucleate cell large enough for the payload to pass through, wherein a width of the constriction is less than 4 micrometers; and incubating the anucleate cell in a solution comprising the payload for a predetermined time before or after the anucleate cell passes through the constriction.

28 . (canceled)

29 . The method of claim 27 , wherein said cell is one or more of red blood cells, erythrocytes, reticulocytes, and platelets.

30 . The method of claim 27 , wherein said cell is a healthy cell.

31 . The method of claim 27 , wherein said cell is an infected cell or a diseased cell.

32 . The method of claim 27 , wherein the cell suspended in a buffer includes unmodified blood.

33 . The method of claim 27 , wherein said width is between 0.5 micrometers and 4 micrometers.

34 . The method of claim 33 , wherein the width is between 2 micrometers and 3 micrometers.

35 . The method of claim 33 , wherein the width is less than the largest diameter of the cell.

36 . The method of claim 33 , wherein the width is about 20% to about 99% the largest diameter of the cell.

37 - 38 . (canceled)

39 . The method of claim 27 , wherein said buffer is a hypotonic buffer that causes said cell to swell.

40 . The method of claim 27 , wherein said payload-containing solution comprises one or more of proteins, peptides, small molecules, nucleic acids, lipids, carbohydrates, macromolecules, vitamins, polymers, fluorescent dyes, fluorophores, carbon nanotubes, quantum dots, nanoparticles, dextran polymers, and steroids.

41 - 45 . (canceled)

46 . The method of claim 27 , wherein a cross-section of the microfluidic channel is selected from the group consisting of circular, elliptical, an elongated slit, square, hexagonal, and triangular.

47 . The method of claim 27 , wherein incubating the cell in a payload-containing solution comprises incubating the cell for 0.0001 seconds to 60 minutes.

48 . A method of treating an infection or disease in a patient in need thereof, comprising:
passing an anucleate cell suspended in a buffer through a microfluidic channel that includes a cell-deforming constriction causing perturbations of the anucleate cell large enough for a payload to pass through, wherein a width of the constriction is less than 4 micrometers; incubating the anucleate cell in a solution comprising the payload for a predetermined time before or after the anucleate cell passes through the constriction; waiting for a predetermined amount of time for the perturbations of the anucleate cell to close such that the payload is contained intracellularly; and administering the anucleate cell to the patient.

49 - 74 . (canceled)

75 . The method of claim 27 , wherein said length is 30 micrometers or less.

76 . The method of claim 27 , wherein said depth is between 1 micrometer and 1 millimeter.

77 . The method of claim 27 , wherein the microfluidic channel comprises a plurality of constrictions.

78 . The method of claim 77 , wherein the plurality of constrictions are arranged in series and/or parallel.

79 . The method of claim 78 , wherein the plurality of constrictions are arranged in parallel.

Assignees

• Massachusetts Inst Technology

Inventors

• Sharei Armon R
• Jensen Klavs F
• Abshire James Robbins
• Niles Jacquin Clarence

Classifications

• C12N5/0641

  Erythrocytes · CPC title

• C12M23/16 Primary

  Microfluidic devices; Capillary tubes (integrated microfluidic structures B01L3/5027; microreactors B01J19/0093) · CPC title

• C12N2521/00

  Culture process characterised by the use of hydrostatic pressure, flow or shear forces · CPC title

• C12N2527/00

  Culture process characterised by the use of mechanical forces, e.g. strain, vibration · CPC title

• C12N5/0644

  Platelets; Megakaryocytes · CPC title