Bubble-free air interchanger of bioreactor

Abstract

The utility model discloses a bioreactor bubble-free air interchanger which comprises a shell, an air inlet pipe extends into the shell, an air outlet port of the air inlet pipe extends into the shell, the top of the shell is connected with an exhaust pipe, an air inlet port of the exhaust pipe is located at the top of the shell, a liquid passing hole is formed in the cylinder wall of the shell, and the liquid passing hole is communicated with the air inlet port of the air inlet pipe. A filtering body for liquid to pass through is arranged on the periphery of the shell and is of a porous net structure. Compared with the prior art, the bubble-free air interchanger of the bioreactor has the advantages that the bubble-free air interchanger of the bioreactor is made of 316 stainless steel materials, is small in occupied space and easy to install and operate, can achieve bubble-free air interchange in the bioreactor, eliminates bubble damage to cells while meeting the requirement of a culture system for oxygen concentration, and improves the air interchange efficiency of the bioreactor. The device is suitable for the culture process of mammalian cells sensitive to shear force. The device is flexible in application, can be adapted to bioreactors of different specifications, and is easy for process development and amplification.

Description

Technical Field

[0001] The utility model relates to an auxiliary device of a bioreactor, in particular to a bubble-free ventilation device of a bioreactor. Background Art

[0002] Bioreactors, essential equipment in the development and production of biological products, provide an optimal environment for cell culture and metabolite synthesis. These environmental parameters include, but are not limited to, temperature, pH, dissolved oxygen (DO), carbon dioxide partial pressure (pCO2), osmotic pressure, and shear stress. The constancy and uniformity of these environmental parameters are often maintained through stirring and aeration. Traditional aeration involves introducing gas through a gas pipe with multiple holes (also known as a gas distributor). This process generates significant shear forces as bubbles rise and break at the gas-liquid interface. Mammalian cells, lacking cell walls, are sensitive to shear forces, negatively impacting cell viability and product yield. Therefore, there is a need to improve existing aeration methods and develop a bubble-free aeration device for bioreactors, a pressing issue for those skilled in the art. Summary of the Invention

[0003] In order to solve the above-mentioned shortcomings, the utility model provides a bubble-free ventilation device for a bioreactor, which occupies a small volume, is easy to install and operate, eliminates bubble damage to cells while meeting the oxygen concentration requirements of the culture system, and is suitable for mammalian cell culture processes that are sensitive to shear forces.

[0004] The above-mentioned purpose of the present utility model is achieved through the following technical solutions: a bubble-free ventilation device for a bioreactor, comprising a shell, characterized in that: an air inlet pipe is extended into the shell, the air outlet port of the air inlet pipe extends into the interior of the shell, the top of the shell is connected to an exhaust pipe, the air inlet port of the exhaust pipe is located at the top of the shell, a liquid hole is provided on the cylindrical wall of the shell, and a filter body for liquid to pass through is provided on the outer periphery of the shell, the filter body has a porous mesh structure, and its function is to allow liquid to circulate, and bubble-free ventilation is achieved through this device, which meets the oxygen concentration requirements of the culture system while eliminating bubble damage to cells.

[0005] Furthermore, the shell includes a top cover, a cylinder and a bottom cover, and the top cover and the bottom cover are respectively connected to the upper and lower ends of the cylinder, and can be connected by welding, threaded connection or integral molding.

[0006] Furthermore, the cylinder is in the shape of a circular tube.

[0007] Furthermore, the bottom cover is a stainless steel sheet.

[0008] Furthermore, the filter body is a stainless steel wire mesh with a welding seam of 2-5 mm in the form of an opaque weld mark. The pore size of the metal mesh can be designed according to different mass transfer capacity requirements, ranging from 10-300 microns.

[0009] Furthermore, the shell is made of 316 stainless steel.

[0010] The bubble-free ventilation device can be designed in different sizes to adapt to bioreactors of different specifications (0.5-15L), and is mainly designed based on parameters such as the total length of the device, bottom diameter, and filter body length.

[0011] The method for using the bubble-free ventilation device comprises the following steps:

[0012] S1: Place the bubble-free ventilation device in the bioreactor and connect the air inlet and outlet pipes to keep the air passages unobstructed. Without introducing any gas, the liquid levels in the bioreactor and the bubble-free ventilation device are level, and the air pressure in the bubble-free ventilation device is equal to the air pressure in the bioreactor.

[0013] S2: Keeping the gas outlet passage unobstructed, introducing gas through the bubble-free ventilation device, partially dissolving the gas in the reaction liquid in the bioreactor, causing the reaction liquid in the bubble-free ventilation device to rise;

[0014] S3: The gas outlet path is still kept unobstructed, the pressure in the bubble-free ventilation device is reduced, and the reaction liquid in the bioreactor enters the bubble-free ventilation device through the filter body until the pressure in the bubble-free ventilation device and the bioreactor is equal and the liquid level is level.

[0015] The advantages of this invention over existing technologies include: Made of 316 stainless steel, the device occupies a small footprint and is easy to install and operate. It achieves bubble-free ventilation within the bioreactor, meeting the culture system's oxygen concentration requirements while eliminating bubble damage to cells. It is suitable for shear-sensitive mammalian cell culture processes. The device is flexible and adaptable to bioreactors of varying sizes, facilitating process development and scale-up. BRIEF DESCRIPTION OF THE DRAWINGS

[0016] Figure 1 It is a structural diagram of the present utility model. DETAILED DESCRIPTION

[0017] The present invention will be further described below in conjunction with the accompanying drawings.

[0018] like Figure 1As shown, a bubble-free ventilation device for a bioreactor includes a shell 1, which is made of 316 stainless steel. The shell 1 includes a top cover 101, a cylinder 102, and a bottom cover 103. The top cover 101 and the bottom cover 103 are respectively connected to the upper and lower ends of the cylinder 102, and can be connected by any method such as welding, threaded connection, or integral molding. The cylinder 102 is in the shape of a circular tube, and the bottom cover 103 is a stainless steel sheet. An air inlet pipe 2 extends into the top cover 101 of the shell 1, and the air outlet port of the air inlet pipe 2 extends into the interior of the shell 1. The top of the shell 1 is connected to an exhaust pipe 3, and the air inlet port of the exhaust pipe 3 is located at the top of the shell 1, at most flush with the bottom side of the top cover 101 of the shell 1, and does not extend into the interior space of the shell 1, so as to facilitate the discharge of gas. The sidewall of the cylindrical body 102 of the housing 1 is provided with a liquid passage hole 4, and the outer periphery of the housing 1 is provided with a filter 5 for liquid passage. The filter 5 is made of stainless steel mesh with a 2-5 mm weld seam, which is in the form of an impermeable weld mark. The pore size of the stainless steel mesh can be designed to meet different mass transfer requirements, ranging from 10 to 300 microns. The filter 5 has a porous mesh structure, which functions to circulate liquid and achieve bubble-free ventilation through this device, meeting the culture system's required oxygen concentration while eliminating bubble damage to cells.

[0019] During use, the bubble-free ventilation device can be docked with the interface of a bioreactor, with the hollow inlet pipe connected to the bioreactor's gas supply line, and the hollow outlet pipe connected to the bioreactor's exhaust line. When gas is introduced, bubbles rise within the device and are discharged through the exhaust pipe. During this process, gas components within the bubbles whose concentrations exceed the equilibrium solubility of corresponding components in the reaction solution enter the reaction solution through mass transfer. Throughout this process, the bubbles do not directly enter the bioreactor, thus preventing shear damage to cells caused by the bubbles rising, coalescing, and rupturing in the reaction solution.

[0020] Application Example 1: Culture Method of Induced Pluripotent Stem Cells (iPSCs):

[0021] In this example, a 0.5L stirred-tank bioreactor with a matching, bubble-free aerator was used, resulting in a working volume of 0.3L. iPSCs were seeded at a density of 5E+04 cells / mL. Starting on day 3, 50% of the medium (0.15L) was exchanged daily, and culture was terminated on day 10. The final cell density reached 1E+06 cells / mL, with cells expanding 20-fold and forming uniformly sized cell clusters. Throughout the entire culture cycle, no bubbles were observed in the reactor, and the exhaust gas path remained unobstructed. Among them, the process parameters are automatically controlled by the bioreactor, the stirring speed is 80-140rpm, and it is increased during the culture process according to the deposition of cell clusters; the temperature is 37°C, and water-free temperature control is achieved by semiconductor Peltier; the DO is 20%, and air and oxygen are introduced through a bubble-free ventilation device, and the gas flow rate is controlled by mass flow meters in different gas paths; the pH is 7.4, and carbon dioxide is introduced through a bubble-free ventilation device, and the gas flow rate is controlled by a mass flow meter. At the same time, it is controlled by the addition of 0.5 mol / L sodium hydroxide solution, and the flow acceleration rate is controlled by the corresponding peristaltic pump.

[0022] Application Example 2: Culture Method of Mesenchymal Stem Cells (MSCs):

[0023] In this example, a 1.5L stirred-tank bioreactor with a matching, bubble-free aerator was used, resulting in a working volume of 0.9L. Cytiva's Cytodex3 microcarriers were used at a microcarrier density of 10g / L. MSC cells were seeded at a density of 1E+05 cells / mL. Starting on day 3, 50% of the medium (0.45L) was exchanged daily, and culture was terminated on day 8. The final cell density was 1.5E+06 cells / mL, representing a 15-fold cell expansion. Throughout the entire culture cycle, no bubbles appeared in the reactor, and the exhaust gas path remained unobstructed. Among them, the process parameters are automatically controlled by the bioreactor, the stirring speed is 60-100rpm, and it is increased during the culture process according to the deposition of microcarriers; the temperature is 37°C, and water-free temperature control is achieved by semiconductor Peltier; the DO is 10%, and air and oxygen are introduced through a bubble-free ventilation device, and the gas flow rate is controlled by mass flow meters in different gas paths; the pH is 7.4, and carbon dioxide is introduced through a bubble-free ventilation device, and the gas flow rate is controlled by a mass flow meter. At the same time, it is controlled by the addition of 0.5 mol / L sodium hydroxide solution, and the flow acceleration rate is controlled by the corresponding peristaltic pump.

[0024] Application Example 3: Culture Method of Monkey Kidney Cells (Vero):

[0025] In this example, a 3L stirred tank bioreactor and a bubble-free aerator of matching size were used with a working volume of 1.5L. Cytiva's Cytodex1 microcarriers were used, the microcarrier density was 5g / L, the Vero cell seeding density was 3E+05 cells / mL, and the culture was terminated on the 6th day with a final cell density of 3E+06 cells / mL, resulting in a 10-fold cell expansion. During the entire culture cycle, no bubbles appeared in the reactor, and the exhaust gas path remained unobstructed. The process parameters were automatically controlled by the bioreactor. The stirring speed was 50-80 rpm, which was increased during the culture process according to the deposition of microcarriers. The temperature was 37°C, and water-free temperature control was achieved by a heating blanket. The DO was 60%, and air and oxygen were introduced through the bubble-free aerator, with gas flow rates controlled by mass flow meters in different gas paths. The pH was 7.2, and carbon dioxide was introduced through the bubble-free aerator, with gas flow rates controlled by mass flow meters. The flow rate was also controlled by the addition of 0.5 mol / L sodium hydroxide solution, with the corresponding peristaltic pump controlling the flow rate.

[0026] The above description is merely an embodiment of the present invention and does not limit the patent scope of the present invention. Any equivalent structure or equivalent process transformation made by using the contents of the description and drawings of the present invention, or directly or indirectly applied in other related technical fields, are also included in the patent protection scope of the present invention.

Claims

1. A bubble-free ventilation device for a bioreactor, comprising a housing, characterized in that: An air inlet pipe extends into the shell, and the air outlet port of the air inlet pipe extends to the interior of the shell. The top of the shell is connected to an exhaust pipe, and the air inlet port of the exhaust pipe is located at the top of the shell. A liquid hole is provided on the cylindrical wall of the shell, and a filter body for liquid to pass through is provided on the outer periphery of the shell, and the filter body has a porous mesh structure.

2. A bioreactor bubble-free ventilation device according to claim 1, characterized in that: The shell includes a top cover, a cylinder and a bottom cover, and the top cover and the bottom cover are respectively connected to the upper and lower ends of the cylinder, and are connected by welding, thread connection or integral molding.

3. A bioreactor bubble-free ventilation device according to claim 2, characterized in that: The cylinder is in the shape of a circular tube.

4. A bioreactor bubble-free ventilation device according to claim 2, characterized in that: The bottom cover is a stainless steel sheet.

5. The bubble-free ventilation device for a bioreactor according to claim 1, characterized in that: The filter body is a stainless steel wire mesh.

6. A bioreactor bubble-free ventilation device according to claim 5, characterized in that: The pore size of the stainless steel wire mesh ranges from 10 to 300 microns.

7. The bubble-free ventilation device for a bioreactor according to claim 1, characterized in that: The shell is made of 316 stainless steel.

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