Devices, systems and methods for automated cell culturing
View Patent ↗A bioreactor is provided. The bioreactor is a multi-scalable bioreactor, which comprises a culture vessel for seeding and culturing cells by adding a cell-culture media, wherein the culture vessel comprises at least a side wall and a bottom surface, a specific heat transfer area and a specific gas transfer area; wherein the culture vessel is configured to accommodate the cell-culture media volume up to 10 liters, and wherein the specific heat transfer area and the specific gas transfer area are independent of cell-culture media volume. A kit for culturing cells in a large scale is also provided which further comprises disposable tubings, culture bag or combinations thereof. A method for culturing cells is also provided.
1. A bioreactor, comprising:
a culture vessel for seeding and culturing cells by adding a cell-culture medium,
wherein the culture vessel comprises at least a side wall and a bottom surface;
wherein the culture vessel is configured such that a specific heat transfer area (sHTA) and a specific gas transfer area (sGTA) of the culture vessel are constant with respect to a change in a volume of the cell-culture medium in the culture vessel, and
wherein the specific heat transfer area at a height h of the cell-culture medium in the culture vessel is defined by equation (1) and the specific gas transfer area at a height h of the cell-culture medium in the culture vessel is defined by equation (2):
sHTA( h )=HTA( h )/ V ( h ) equation (1), and
sGTA( h )=GTA( h )/ V ( h ) equation (2),
wherein sHTA(h) is a specific heat transfer area for a height h of the cell-culture medium in the culture vessel, HTA(h) is a heat transfer area for the height h of the cell-culture medium in the culture vessel, V(h) is a volume of the cell-culture medium for the height h of the cell-culture medium in the culture vessel, sGTA(h) is a specific gas transfer area at the height h of the cell-culture medium in the culture vessel, and GTA(h) is an area of a top surface of the cell-culture medium at the height h of the cell-culture medium in the culture vessel.
2. The bioreactor of claim 1 , wherein the culture vessel is configured to accommodate the volume of cell-culture medium between 15 ml to 10 liters.
3. The bioreactor of claim 1 , wherein the culture vessel is configured to accommodate an expandable cell-culture medium volume.
4. The bioreactor of claim 1 , wherein the bioreactor is a multi scale bioreactor.
5. The bioreactor of claim 1 , wherein the bioreactor system is configured to maintain a set point for constant temperature, pH, dissolved oxygen or combinations thereof during operation.
6. The bioreactor of claim 1 , further comprising a controller for controlling a gas mixing ratio, gas flow rate, heating, agitating or combinations thereof within the culture vessel.
7. The bioreactor of claim 1 , further comprising one or more impellers for agitating the culture medium.
8. The bioreactor of claim 7 , wherein the impellers are operably coupled to a motor by one or more of magnetic coupling, mechanical coupling, electrical coupling and electromagnetic coupling.
9. The bioreactor of claim 7 , wherein the impellers are operably coupled to a motor by magnetic coupling.
10. The bioreactor of claim 1 , further comprising one or more heating units for heating the culture medium.
11. The bioreactor of claim 10 , wherein at least one of the heating units is used to heat the side-walls.
12. The bioreactor of claim 10 , wherein the heating unit comprises an IR source, a heating blanket, a water jacket or combinations thereof.
13. The bioreactor of claim 1 , further comprising one or more temperature controllers for maintaining the temperature of the culture medium.
14. The bioreactor of claim 1 , further comprising a bioreactor lid.
15. The bioreactor of claim 14 , further comprising one or more heating units for heating the bioreactor lid.
16. The bioreactor of claim 14 , further comprising one or more temperature controllers to control a temperature of the bioreactor lid.
17. The bioreactor of claim 1 , further comprising one or more temperature sensors for providing a temperature feedback control of the culture medium.
18. The bioreactor of claim 1 , further comprising one or more pH sensors and/or dissolved oxygen sensors.
19. The bioreactor of claim 1 , wherein the culture vessel has a length in a range of 1 to 500 cm, a width in a range of 1 to 500 cm and a height in a range of 1 to 100 cm.
20. The bioreactor of claim 1 , wherein the culture vessel has an aspect ratio in a range from about 0.3 to about 3.
21. The bioreactor of claim 1 , wherein the culture vessel is made of glass, polymer, ceramic, metal, or a combination thereof.
22. The bioreactor of claim 1 , wherein the culture vessel is made of thermoplastic.
23. The bioreactor of claim 1 , wherein the culture vessel has a perimeter that is triangular, rectangular, square planar, pentagonal, hexagonal, polygonal, circular, elliptical, or irregular.
24. The bioreactor of claim 1 , further comprising a biocompatible coating on an inner wall of the culture vessel, wherein the biocompatible coating comprises notch ligands, anti-CD3 antibody and anti-CD28 antibody.
25. The bioreactor of claim 1 is a stirred tank bioreactor, a reactor with a rocking or rolling motion, a reactor with a one dimensional shaking, a reactor with two dimensional shaking, a reactor with three dimensional shaking, or combinations thereof.
26. The bioreactor of claim 1 is configured to seed and culture cells selected from bacterial cells, animal cells, plant cells, fungi, insect cells, microbes, virus cells, stem cells, or combinations thereof.
27. The bioreactor of claim 1 , further comprising one or more conduits.
28. The bioreactor of claim 27 , wherein the conduits are used for adding culture medium, nutrients, cells, gases, acid, base, or combinations thereof to the vessel, and/or the conduits are used for withdrawing the culture media from the vessel for sampling, transferring the cell culture media with cells to a cell-harvester or combinations thereof under sterile conditions.
29. The bioreactor of claim 1 , wherein the bioreactor further comprises one or more gas inlet ports and/or gas outlet ports.
30. The bioreactor of claim 1 , wherein the gas inlet ports and/or gas outlet ports comprise a sterile filter.
31. The bioreactor of claim 30 , wherein the gas outlet ports comprise an off-gas condenser.
32. The bioreactor of claim 31 , wherein the gas inlet ports comprise a heater.
33. A kit comprising:
one or more disposable tubings; and
a bioreactor comprising:
a culture vessel for seeding and culturing cells by adding a cell-culture medium, wherein the culture vessel comprises at least a side wall and a bottom surface, wherein the culture vessel is configured to accommodate the cell-culture medium without introducing a significant change in bioreactor dynamics by maintaining a specific heat transfer area and a specific gas transfer area,
wherein the culture vessel is configured such that a specific heat transfer area (sHTA) and a specific gas transfer area (sGTA) of the culture vessel are constant with respect to a change in a volume of the cell-culture medium in the culture vessel; and the specific heat transfer area and a specific gas transfer area at a height h of the cell-culture medium in the culture vessel are defined by equations (1) and (2), respectively:
sHTA ( h )=HTA ( h )/ V ( h ) equation (1)
sGTA ( h )=GTA ( h )/ V ( h ) equation (2),
wherein sHTA(h) is a specific heat transfer area for a height h of the cell-culture medium in the culture vessel, HTA(h) is a heat transfer area for the height h of the cell-culture medium in the culture vessel, V(h) is a volume of the cell-culture medium for the height h of the cell-culture medium in the culture vessel, sGTA(h) is a specific gas transfer area at the height h of the cell-culture medium in the culture vessel, and GTA(h) is an area of a top surface of the cell-culture medium at the height h of the cell-culture medium in the culture vessel.
34. The kit of claim 33 further comprising a non-disposable pump or a disposable pump.
35. The bioreactor of claim 1 , wherein the culture vessel has a wavy perimeter.
36. The bioreactor of claim 35 , wherein the wavy perimeter varies with respect to a change in the height or volume of the culture vessel.
37. The bioreactor of claim 1 , wherein the culture vessel has a circular base, and a height (h k ) represented by a function h k =h 0 +k·δh, and a radius at the height (h k ) is represented by a function
r
k
=
r
0
·
(
h
0
h
0
-
δ
h
)
k
/
2
,
and
wherein the culture vessel has a flower-like shape having a perimeter P(h) at a height h defined as:
HTA
(
h
k
)
=
∫
0
h
P
(
h
)
sin
(
θ
h
)
ⅆ
h
≈
2
π
r
0
h
0
+
∑
i
=
1
k
a
i
π
(
r
i
+
r
i
-
1
)
sin
θ
i
δ
h
wherein an angle between the side wall of the culture vessel and a gas transfer surface at the height h is defined as θ h ,for h greater than h 0 , wherein r k and r 0 are radii of cross-sectional areas of the culture vessel at heights h k and h 0 , respectively, wherein a i is a scaling factor of the perimeter at i th height, and wherein a i can be computed recursively, such that the ratio of the computed HTA(h k ) and V(h k ) is constant for the height h k of the cell-culture medium.
38. The bioreactor of claim 33 , wherein the culture vessel is configured to accommodate the cell-culture medium having a volume in a range from 10 ml to 10 liters.