AUTOMATIC IN VITRO CELL CULTURE PLATFORM AND CELL CULTURE METHOD

• US 20200131466A1
• Filed: 11/16/2018
• Published: 04/30/2020
• Est. Priority Date: 10/30/2018
• Status: Active Grant

• Current Assignee: National Tsing Hua University
• Original Assignee: National Tsing Hua University
• Inventors: Hsieh, Hsin-Lin, Huang, Jen-Huang
• Granted Patent: US 11,371,009 B2
• Time in Patent Office: Days
• CPC Class Codes: C12M 23/40 Manifolds; Distribution pieces fluid transfer means B01L3/563
• CPC Class Codes: C12M 25/02 Membranes; Filters filters or filtration in general B01D24/00-B01D41/00
• CPC Class Codes: C12M 25/06 Plates; Walls; Drawers; Multilayer plates
• CPC Class Codes: C12M 29/00 Means for introduction, extraction or recirculation of materials, e.g. pumps pumps per se F04B
• CPC Class Codes: C12M 29/04 Filters; Permeable or porous membranes or plates, e.g. dialysis
• CPC Class Codes: C12M 41/48 Automatic or computerized control automatic analysis G01N35/00

Abstract

An automatic in vitro cell culture platform includes a fluid storage device, a channel control panel, a cell culture chip and a fluid drive unit. The fluid storage device is for storing a cell culture medium and a waste liquid. The channel control panel is pipe-connected to the fluid storage device. The cell culture chip is disposed on the channel control panel and is for culturing a cell. The fluid drive unit is pipe-connected to the channel control panel and is for driving the cell culture medium and the waste liquid to flow between the fluid storage device and the cell culture chip through the channel control panel.

Claims

1. An automatic in vitro cell culture platform, comprising:

a fluid storage device, comprising;

a first fluid storage unit for storing a cell culture medium; and

a second fluid storage unit disposed adjacent to the first fluid storage unit and for storing a waste liquid;

a channel control panel pipe-connected to the fluid storage device and comprising a valve group, wherein the valve group is for controlling a transport of the cell culture medium and the waste liquid;

a cell culture chip disposed on the channel control panel and pipe-connected thereto, and the cell culture chip comprising;

an upper chamber for culturing a cell;

a lower chamber disposed under the upper chamber and for accommodating the cell culture medium; and

a porous semipermeable membrane disposed between the upper chamber and the lower chamber, wherein the cell culture medium can flow between the upper chamber and the lower chamber through the porous semipermeable membrane, and a liquid level of the cell culture medium in the cell culture chip is higher than a level of the porous semipermeable membrane in the cell culture chip; and

a fluid drive unit pipe-connected to the channel control panel and for driving the cell culture medium and the waste liquid to flow between the fluid storage device and the cell culture chip through the channel control panel.

2. The automatic in vitro cell culture platform of claim 1, wherein the cell culture chip has a pipe connection surface and comprises, in order from the pipe connection surface to a bottom of the cell culture chip:

a first base plate;

a second base plate;

a third base plate comprising an opening, wherein the porous semipermeable membrane is disposed and covered on the opening;

a fourth base plate; and

a resin sheet;

wherein the first base plate, the second base plate and the third base plate are sequentially stacked so as to form the upper chamber, and the third base plate, the fourth base plate and the resin sheet are sequentially stacked so as to form the lower chamber.

3. The automatic in vitro cell culture platform of claim 2, wherein the cell culture chip further comprises:

at least four pipe ports separately disposed on the pipe connection surface, wherein two of the pipe ports are connected to the upper chamber, and the other two of the pipe ports are connected to the lower chamber; and

at least four infusion tubes, wherein one end of each of the infusion tubes is connected correspondingly to one of the pipe ports, and the other end of each of the infusion tubes is connected to the channel control panel.

4. The automatic in vitro cell culture platform of claim 2, wherein the first base plate, the second base plate, the third base plate, the fourth base plate and the resin sheet are made by a laser cutting method.

5. The automatic in vitro cell culture platform of claim 1, wherein the valve group comprises a plurality of valve elements for controlling the transport of the cell culture medium and the waste liquid, respectively.

6. The automatic in vitro cell culture platform of claim 1, wherein the channel control panel has a tube connection surface and comprises, in order from the tube connection surface to a bottom of the channel control panel:

a tube connection plate;

a first channel base plate;

a second channel base plate;

a third channel base plate; and

a bottom plate;

wherein the tube connection plate, the first channel base plate, the second channel base plate, the third channel base plate and the bottom plate are sequentially stacked so as to form a plurality of microchannels for transporting the cell culture medium and the waste liquid.

7. The automatic in vitro cell culture platform of claim 6, wherein the channel control panel further comprises:

a plurality of channel ports separately disposed on the tube connection surface; and

a plurality of connecting tubes, wherein one end of each of the connecting tubes is connected correspondingly to one of the channel ports, and the other end of each of the connecting tubes is connected to the fluid drive unit or the fluid storage device.

8. The automatic in vitro cell culture platform of claim 6, wherein the tube connection plate, the first channel base plate, the second channel base plate, the third channel base plate and the bottom plate are made by a laser cutting method.

9. The automatic in vitro cell culture platform of claim 1, wherein the fluid storage device further comprises a medium exchange port and a waste liquid exchange port, the medium exchange port is connected to the first fluid storage unit, and the waste liquid exchange port is connected to the second fluid storage unit.

10. The automatic in vitro cell culture platform of claim 1, wherein the fluid drive unit is a peristaltic pump.

11. The automatic in vitro cell culture platform of claim 1, wherein an average pore size of the porous semipermeable membrane ranges from 0.2 μ

m to 8 μ

m.

12. The automatic in vitro cell culture platform of claim 1, wherein the cell culture medium comprises a growth factor and a cell differentiation factor.

13. The automatic in vitro cell culture platform of claim 1, wherein the first fluid storage unit comprises:

a plurality of fluid storage subunits separately disposed in the first fluid storage unit.

14. The automatic in vitro cell culture platform of claim 1, further comprising:

a basal board for loading the fluid storage device, the channel control panel, the channel control panel, the cell culture chip and the fluid drive unit.

15. A cell culture method, comprising:

providing the automatic in vitro cell culture platform of claim 1;

performing a washing step, wherein the cell culture medium is transported from the first fluid storage unit to the cell culture chip by the fluid drive unit so as to wash the upper chamber and the lower chamber, and then the cell culture medium used to wash the upper chamber and the lower chamber is transported from the cell culture chip to the second fluid storage unit by the fluid drive unit;

performing a cell culture medium filling step, wherein the cell culture medium is transported from the first fluid storage unit to the cell culture chip by the fluid drive unit so as to fill the cell culture medium into the upper chamber and the lower chamber, respectively;

performing a cell seeding step, wherein the cell is seeded in the upper chamber for an incubation time, and the cell is attached and grows on the porous semipermeable membrane; and

performing a cell culture medium replacing step, wherein the cell culture medium cultured after the incubation time is removed from the cell culture chip by the fluid drive unit, and then the cell culture medium of the first fluid storage unit is transported from the first fluid storage unit to the cell culture chip by the fluid drive unit.

16. The cell culture method of claim 15, wherein the cell culture medium replacing step comprises:

performing an upper-chambered cell culture medium removing step, wherein the cell culture medium cultured after the incubation time is transported from the upper chamber to the second fluid storage unit by the fluid drive unit, and then the cell culture medium of the first fluid storage unit is transported to the cell culture chip by the fluid drive unit; and

performing a cell culture medium exchanging step, wherein the cell culture medium of the cell culture chip is exchanged with the cell culture medium of the first fluid storage unit by the fluid drive unit.

17. The cell culture method of claim 15, further comprising:

repeating the cell culture medium replacing step for elongating a survival time of the cell in the cell culture chip.

18. The cell culture method of claim 15, further comprising:

performing a cell differentiation inducing step, wherein a growth factor and a cell differentiation factor are added into the cell culture medium, and the cell culture medium comprising the growth factor and the cell differentiation factor is transported from the first fluid storage unit to the cell culture chip by the fluid drive unit so as to induce the cell to differentiate into a functional cell.