Microfluidic bio-reactor device

Patent metadata

Field	Value
Publication number	US-10106766-B2
Application number	US-201514681631-A
Country	US
Kind code	B2
Filing date	Apr 8, 2015
Priority date	Jan 30, 2015
Publication date	Oct 23, 2018
Grant date	Oct 23, 2018

Abstract

The present invention provides a microfluidic bio-reactor device, which comprises: a first cell culture layer; a microfluidic base, which is located on the top of the first cell culture layer; a microfluidic layer, which is located on the top of the microfluidic base and have an air-bubble removal device; a microfluidic roof, which is located on the top of the microfluidic layer; the present invention also provides a method for culturing cells by the microfluidic bio-reactor device of the present invention and the kit with a cell-loading device and present microfluidic bio-reactor device.

First claim

What is claimed is:

1. A microfluidic bio-reactor device comprising:
a first cell culture layer comprising an opening defined by a continuous boundary forming the opening, said first cell culture layer located on a top of a first cell culture substrate, wherein said first cell culture layer and said first cell culture substrate form an inside of a first cell culture chamber;
a microfluidic base, the microfluidic base is located on a top of the first cell culture layer and has a first channel, a second channel and holes, said first channel and said second channel are located inside an area corresponding to the inside of the first cell culture chamber, said holes are located inside an area corresponding to the first cell culture layer;
a microfluidic layer, the microfluidic layer is located on a top of the microfluidic base, and the microfluidic layer comprising:
a gas permeable layer;
a curved channel;
an inlet having a first end and a second end, the first end of the inlet being an opening and the second end of the inlet being connected to the curved channel;
a first diffusion device having a first end and a second end, the first end of the first diffusion device being connected to the curved channel and the second end of the first diffusion device being connected to the gas permeable layer, said first diffusion device has an opening which is connected to the first channel of the microfluidic base;
an air-bubble removal device having a first end and a second end, the air-bubble removal device being a vacuum channel, the first end of the air-bubble-removal device being connected to the gas permeable layer and the second end being connected to a vacuum opening, said vacuum channel is connected to the holes of the microfluidic base;
an outlet having a first end and a second end, the first end of the outlet being an opening and the second end being connected to a second diffusion device, said second diffusion device has an opening which is connected to the second channel of the microfluidic base; and
a microfluidic roof, the microfluidic roof is located on a top of the microfluidic layer.

2. The microfluidic bio-reactor device of claim 1 , wherein the first diffusion device and the second diffusion device are fan-shaped.

3. The microfluidic bio-reactor device of claim 1 , wherein the inlet and the outlet are at opposite sides of the microfluidic layer.

4. The microfluidic bio-reactor device of claim 1 , wherein the first cell culture layer, the microfluidic layer and the gas permeable layer are polydimethylsiloxane (PDMS).

5. The microfluidic bio-reactor device of claim 1 , wherein an area of the first cell culture substrate is bigger than the area of the first cell culture layer.

6. The microfluidic bio-reactor device of claim 5 , wherein the first cell culture substrate is culture dish or slide.

7. The microfluidic bio-reactor device of claim 1 , which further used with an adhesion material and a vacuum tube connected with a vacuum device.

8. The microfluidic bio-reactor device of claim 1 , which further comprising:
a second cell culture layer, the second cell culture layer is located on a top of the microfluidic roof and comprising an opening defined by a continuous boundary forming the opening, wherein said second cell culture layer and said microfluidic roof form an inside of a second cell culture chamber; and
a second cell culture substrate, the second cell culture substrate is located on a top of the second cell culture layer, an area of the second cell culture substrate is bigger than an area of the second cell culture layer;
wherein the microfluidic roof further comprises a third channel, a fourth channel and holes, the third channel and the fourth channel are located inside an area corresponding to the inside of the second cell culture chamber, the holes are located on the second cell culture layer, the holes are connected to the vacuum channel of the microfluidic layer.

9. The microfluidic bio-reactor device of claim 8 , wherein the second cell culture layer is polydimethylsiloxane (PDMS).

10. The microfluidic bio-reactor device of claim 8 , wherein the second cell culture substrate is culture dish or slide.

11. The microfluidic bio-reactor device of claim 1 , wherein the vacuum opening is further connected to a vacuum device.

12. A method for culturing cells by microfluidic bio-reactor device comprising:
(1) seeding cells evenly on the first cell culture substrate of the microfluidic bio-reactor device of claim 1 ;
(2) assembling the microfluidic bio-reactor device;
(3) connecting the vacuum opening of the microfluidic bio-reactor device with a vacuum device to generate a negative pressure making the first cell culture layer and first cell culture substrate adhered tightly;
(4) the vacuum device continuously generating negative pressure to maintain the air-bubble removal device as vacuum status;
(5) injecting a culture medium from the inlet, the culture medium flowing through the opening of the first diffusion device and the first channel of the microfluidic base into the first cell culture layer, wherein air-bubble in the culture medium would pass the gas permeable membrane into the air-bubble removal device;
(6) the culture medium in the first cell culture layer would flow through the second channel of the microfluidic base and the opening of the second diffusion device into the outlet.

13. The method of claim 12 , wherein the cells are stem cells.

14. A kit for culturing cells comprising:
a microfluidic bio-reactor device of claim 1 ;
a vacuum tube connected with a vacuum device; and
an adhesion material.

15. The kit of claim 14 , wherein the adhesion material is double-sided tape or polydimethylsiloxane (PDMS).

Assignees

• Univ Nat Yang Ming

Inventors

• Lee Oscar Kuang-Sheng
• Yen Meng-Hua
• Kuo Shu-Wen

Classifications

• C12M23/24

  Gas permeable parts · CPC title

• C12M29/20

  Degassing; Venting; Bubble traps (means for collection or storage of gas C12M23/36; gas collection apparatus for laboratory use B01L5/02) · CPC title

• C12M23/16 Primary

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

• C12M29/06

  Nozzles; Sprayers; Spargers; Diffusers (per se B01F23/231, B01J19/26) · CPC title