Systems and methods for seeding cell cultures in a microfluidic device

Patent metadata

Field	Value
Publication number	US-11760967-B2
Application number	US-202017013454-A
Country	US
Kind code	B2
Filing date	Sep 4, 2020
Priority date	Sep 5, 2019
Publication date	Sep 19, 2023
Grant date	Sep 19, 2023

Abstract

This disclosure provides systems and methods for seeding cell cultures in a microfluidic device. The systems and methods of this disclosure can enable flow of a cell solution from one side of a scaffold, such as a porous substrate or membrane, to the other side of the scaffold. Flow of the liquid can pass through the scaffold while the cells themselves do not, resulting in the cells driven to the surface of the scaffold for consequent attachment. A microfluidic device can include a microfluidic feature structured to create a seal between a cell seeding tool and an inlet to a microchannel of the microfluidic device. This can enable a pressure-driven flow to push fluid down the channel and through pores of the membrane. In contrast, traditional gravity fed seeding of cells may not create enough pressure to drive fluid through the pores of the scaffold.

First claim

What is claimed is:

1. A microfluidic device, comprising:
a first channel having a first inlet port and a first outlet port, the first channel configured to receive a fluid sample comprising cells, the first inlet port defining a tapered portion that creates a seal between the first inlet port and a cell seeding tool that provides the fluid sample;
a second channel coupled to the first channel via an overlapping region, the second channel having a second inlet port and a second outlet port; and
an overlapping region between the first channel and the second channel, configured to trap the cells in the fluid sample as the fluid sample is forced by the cell seeding tool through the overlapping region via the first channel.

2. The microfluidic device of claim 1 , wherein the overlapping region forms a bottom portion of the first channel and forms a top portion of the second channel.

3. The microfluidic device of claim 1 , wherein at least one of the first inlet port, the first outlet port, the second inlet port, or the second outlet port is coupled to a well of a well plate.

4. The microfluidic device of claim 1 , wherein the overlapping region comprises a semipermeable membrane configured to allow the fluid sample to flow from the first channel to the second channel while trapping the cells in the fluid sample on a surface of the semipermeable membrane in the first channel.

5. The microfluidic device of claim 4 , wherein the overlapping region is further configured to trap cells in a second fluid sample on a second surface of the semipermeable membrane in the second channel while allowing a second fluid sample to flow from the second channel to the first channel.

6. The microfluidic device of claim 1 , wherein the overlapping region comprises at least one of a membrane, a filter, a mesh, or a scaffold.

7. The microfluidic device of claim 1 , wherein one or more portions of the overlapping region are chemically treated by at least one of:
a coating;
an energetic plasma treatment;
affixing the one or more portions with a self-assembled monolayer; or
surface depositing.

8. The microfluidic device of claim 1 , wherein an opening of at least one of the second inlet port, the first outlet port, or the second outlet port is defined by a first width and comprises a second tapered portion defining a second width, and is configured to receive a portion of the cell seeding tool.

9. The microfluidic device of claim 8 , wherein the tapered portion is at least one of a squared tapering or a rounded tapering.

10. The microfluidic device of claim 8 , wherein the tapered portion comprises one or more flanges arranged concentrically with the tapered portion that mechanically flex to accommodate the cell seeding tool.

11. The microfluidic device of claim 8 , wherein the opening of at least one of the the second inlet port, the first outlet port, or the second outlet port comprises at least one of a gasket or a spacer configured to create a seal with the portion of the cell seeding tool.

12. The microfluidic device of claim 1 , wherein an opening of at least one of the first inlet port, the second inlet port, the first outlet port, or the second outlet port comprises at least one of one or more fins, a chamfer, a countersink, or an extension into a well of a well plate.

13. A system, comprising:
a semipermeable membrane having a first surface and a second surface opposite the first surface, each of the first surface and the second surface configured to trap cells in a fluid sample;
a first channel coupled to the semipermeable membrane and having a first portion defined by the first surface of the semipermeable membrane, the first channel comprising an inlet port defining a tapered portion that creates a seal between the inlet port and a cell seeding tool that provides the fluid sample; and
a second channel coupled to the semipermeable membrane and having a second portion defined by the second surface of the semipermeable membrane,
wherein the semipermeable membrane is configured to allow the fluid sample to flow between the first channel and the second channel as the fluid sample is forced by the cell seeding tool through the semipermeable membrane.

14. The system of claim 13 , wherein the semipermeable membrane comprises at least one of a membrane, a filter, a mesh, or a scaffold.

15. The system of claim 13 , wherein the semipermeable membrane is further configured to:
trap, on the first surface, first cells of a first fluid sample flowing from the first channel to the second channel; and
trap, on the second surface, second sells of a second fluid sample flowing from the second channel to the first channel.

16. The system of claim 13 , wherein a portion of the semipermeable membrane is configured to limit cell attachment.

17. The system of claim 13 , wherein at least a portion of the semipermeable membrane is chemically treated by at least one of:
a coating;
energetic plasma treatment;
affixing the one or more surfaces with a self-assembled monolayer; or
surface depositing.

Assignees

• Charles Stark Draper Laboratory Inc

Inventors

• Azizgolshani Hesham
• Cain Brian
• Charest Joseph
• Coppeta Jonathan Robert
• Isenberg Brett
• Petrie Timothy

Classifications

• C12M23/16 Primary

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

• C12M25/02

  Membranes; Filters (filters or filtration in general B01D24/00-B01D41/00) · CPC title

• B01L3/502753

  characterised by bulk separation arrangements on lab-on-a-chip devices, e.g. for filtration or centrifugation · CPC title

• B01L2300/0681

  Filter · CPC title

• B01L3/50255

  Multi-well filtration · CPC title