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Fluidized bed bioreactor

A technology of bioreactor and fluidized bed, which is applied in the direction of bioreactor/fermenter combination, specific-purpose bioreactor/fermenter, biochemical instrument, etc., and can solve the problem of cell and carrier flushing out, limited amplification effect, etc. problem, to achieve the effect of good amplification potential, utilization of tank volume, and convenient extraction and processing

Active Publication Date: 2011-06-29
SHANGHAI RITAI MEDICINE EQUIP PROJECT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention provides a new fluidized bed bioreactor aiming at the defects that the existing fluidized bed bioreactor is easy to cause cells and carriers to wash out and the amplification effect is limited

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0032] Embodiment 1: Utilize the fluidized bed bioreactor of the present invention to carry out Vero cell culture

[0033] In the fluidized bed bioreactor (Shanghai Ritai 10L type), add sheet polyester carrier, add phosphate buffer (pH7.2) at the same time, sterilize under high pressure at 123 ℃ for 60 minutes, and naturally cool overnight. On the second day, the phosphate buffer solution in the tank was drained, and the DMEM medium containing 10% fetal bovine serum was added to inoculate Vero cells. After inoculation, the cell density in the tank reached 5x10 5 / ml to start cell culture.

[0034] Set the reactor cell culture conditions as temperature 37°C, pH 7.2, DO 50% for cell culture. Start the stirring device and set the speed at 80 rpm. At this speed, the speed of the culture solution is lower than the flushing point, and the carrier settles down, forming a culture method similar to fixed bed culture. The working state diagram is shown in image 3 , the carrier is rel...

Embodiment 2

[0038] Embodiment 2: Utilize the fluidized bed bioreactor of the present invention to carry out the mass culture of Chinese hamster ovary cell (CHO cell)

[0039] Add 300 grams of domestic flake polyester carrier and 8 L of phosphate buffer (pH 7.2) into a 10 L fluidized bed bioreactor, autoclave at 123 °C for 60 minutes, and cool overnight naturally. On the second day, the phosphate buffer solution in the tank was drained, and DMEM medium containing 10% fetal bovine serum was added to inoculate CHO cells. After inoculation, the cell density in the tank reached 5x10 5 / ml to start cell culture.

[0040] Set the reactor cell culture conditions as temperature 37°C, pH 7.2, DO 50% for cell culture. Start the stirring device and set the speed at 80 rpm. At this speed, the speed of the culture solution is lower than the flushing point, forming a culture method similar to a fixed bed, but a slight shaking of the carrier can be observed, which helps Homogenize the culture medium an...

Embodiment 3

[0044] Embodiment 3: Utilize the fluidized bed bioreactor of the present invention to carry out the mass culture of 293 cells

[0045] Add 300 grams of domestic flake polyester carrier and 8 L of phosphate buffer (pH 7.2) into a 10 L fluidized bed bioreactor, autoclave at 123 °C for 60 minutes, and cool overnight naturally. On the second day, the phosphate buffer solution in the tank was drained, and high-sugar DMEM medium was added to inoculate 293 cells. After inoculation, the cell density in the tank reached 5x10 5 / ml to start cell culture.

[0046] Set the cell culture conditions of the reactor as temperature 37°C, pH 7.2, DO 50% for cell culture, start the stirring device, and set the speed at 80 rpm. At this speed, the fluid velocity is lower than the flushing point, forming a similar The culture method is a fixed bed, but a slight shaking of the carrier can be observed, which helps to uniform the culture solution, and culture in this state for 1 hour.

[0047] Change...

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Abstract

The invention relates to a biological cell culture device, discloses a fluidized bed bioreactor, which comprises a tank body (1), a honeycomb duct (2), a fixed plate (3), a stirring device (4) disposed in the tank body (1), and a porous round pipe (5) disposed outside the stirring device (4) and connected to the honeycomb duct (2), wherein a plurality of through holes are disposed on the porous round pipe (5) and are connected to the bottom of the tank body (1). The fluidized bed bioreactor of the invention restricts carriers under the fixed plate, which facilitates the extraction and processing of supernatants and the more effective utilization of the tank volume; the fluidized bed bioreactor of the invention combines the advantages of the fluidized bed culture mode and the fixed bed culture mode, realizes a long-time continuous perfusion culture with a homogeneous cell growth environment, a good amplification potential and a cell density being above 107 / ml, and has a good industrial application prospect.

Description

technical field [0001] The invention relates to biological cell culture equipment, in particular to a fluidized bed bioreactor. Background technique [0002] Currently, cell culture methods adopt fluidized bed bioreactors or fixed bed bioreactors for culturing. The fixed bed bioreactor is a fixed container installed in the reactor, and the carrier is filled in the middle. The carrier is usually a solid or porous ball or sheet-like fiber with a diameter of 2-6mm. The culture medium is circulated through the fixed bed, relying on the negative pressure generated during the stirring, so that the medium continuously flows through the filler for the transfer of nutrients and oxygen. The freshly inoculated cells grow on the surface of the carrier, and as the cells proliferate, the cells begin to fill the spaces between the carriers. [0003] The fixed-bed bioreactor has a relatively high flow rate of the culture solution, and the cells grow at a high density in the carrier, which...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12M1/02C12M1/38C12M1/34
CPCC12M27/02C12M23/08C12M27/24C12M25/20
Inventor 郑振彬陈光南
Owner SHANGHAI RITAI MEDICINE EQUIP PROJECT
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