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Centrifugal settling-type bioreactor for continuous culture of plant cells

A technology of bioreactor and centrifugal sedimentation, which is applied in the direction of specific-purpose bioreactor/fermenter, bioreactor/fermenter combination, tissue cell/virus culture device, etc., which can solve the problem of high cost, waste, plant cell The interception rate is not very high, to achieve the effect of preventing pollution and improving efficiency

Inactive Publication Date: 2016-05-25
NANJING XIAOZHUANG UNIV
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0004] However, the plant cell interception rate of the above-mentioned bioreactor is not very high, which will cause a certain waste, because the plant cell itself is difficult to obtain, and the cost is relatively high

Method used

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  • Centrifugal settling-type bioreactor for continuous culture of plant cells

Examples

Experimental program
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Effect test

Embodiment 1

[0018] Such as figure 1 As shown, the centrifugal sedimentation bioreactor for continuous cultivation of plant cells includes a main body 9 with a U-shaped structure as a whole. The main body 9 includes a riser 2 and a downcomer 10. The bottom and top positions are connected respectively; the liquid inlet 1 is also provided on one side of the ascending liquid area 2, and the cell harvesting port 4 is provided at the bottom position of the descending liquid part 10; the ascending liquid part 2 is also provided with a The air nozzle 3, the air nozzle 3 sprays upward airflow, and then drives the culture solution 12 in the ascending liquid part 2 to rise; the descending liquid part 10 is provided with a sealed funnel 6 and a spiral tube 5 from top to bottom, and the top of the sealed funnel 6 is connected to the The inner wall of the downcomer 10 of the main body 9 is bonded and sealed, and the bottom of the sealed funnel 6 is connected to the inlet 51 of the spiral tube 5 . The ...

Embodiment 2

[0025] The method for continuously culturing cells using a bioreactor comprises the steps of: adopting the culture medium to add NAA and IAA MS medium to plant cells in a U-shaped centrifugal sedimentation bioreactor for cultivating, keeping the air nozzle from coming out The aeration rate is 200-300vvm, the culture temperature is 20-30°C, and the culture is 25-35 days.

[0026] In the above embodiment, the reactor includes a main body 9 with a U-shaped structure as a whole, the main body 9 includes a riser 2 and a downcomer 10, and the corresponding bottom and top positions of the riser 2 and the downcomer 10 communicate respectively; A liquid inlet 1 is also provided on one side of the ascending liquid area 2, and a cell harvesting port 4 is provided at the bottom of the descending liquid part 10; The upward airflow is ejected, and then the culture solution 12 in the ascending liquid part 2 is driven to rise; the descending liquid part 10 is provided with a sealed funnel 6 a...

Embodiment 3

[0031] The device used in this example is figure 1shown. Glycyrrhizae cells were continuously cultured through this example, the initial inoculation density was 5 g (dry weight) / L, the culture medium was MS medium supplemented with NAA and IAA, the aeration rate was 300vvm, and the culture temperature was 25°C. At the same time, the cut-off efficiency and kLa of the reactor were measured using licorice cells as the material. The results showed that under conditions suitable for plant cell growth (aeration rate of 0.3vvm, perfusion rate of 10%), the cell retention efficiency reached 100%, the volumetric oxygen dissolution coefficient kLa was 5.0, and the maximum cell biomass reached 14.0g after 27 days / L, higher than the maximum biomass of 12.0g / L in batch culture. The device involved in the invention is very suitable for the continuous cultivation of plant cells and can be popularized and applied on a large scale.

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Abstract

The invention discloses a centrifugal settling-type bioreactor for continuous culture of plant cells. The centrifugal settling-type bioreactor comprises a main body which is in a U-shaped structure on the whole; a liquid inlet is also formed in one side of a liquid lifting region; a cell harvest opening is formed in the bottom position of a liquid dropping part; a liquid lifting part is provided with a gas nozzle at the bottom position of the main body; a seal funnel and a spiral pipe are arranged on the liquid dropping part from top to bottom; the top part of the seal funnel and the inner wall of the liquid dropping part of the main body are in fit seal; the bottom part of the seal funnel is connected to the inlet of the spiral pipe; a gravity settling pipe is also connected to the bottom part of the spiral pipe; and the other side of the gravity settling pipe is connected with a waste liquid collector. Separation of cells and a culture solution is achieved by the gravity settling pipe, which is connected with the bottom of the spiral pipe, in the bioreactor; the cell intercept efficiency can be obviously improved by adding the spiral pipe to the reactor, so that continuous perfusion culture of the plant cells is achieved; and the culture efficiency of the plant cells is improved.

Description

technical field [0001] The invention relates to the technical field of a plant cell culture method and a bioreactor suitable for cultivation, in particular to a centrifugal sedimentation type centrifugal sedimentation type bioreactor for continuous cultivation of plant cells. Background technique [0002] The development of plant natural products has always been a research hotspot. The use of bioreactors for plant tissue culture in vitro can not only produce many valuable secondary metabolites, but also alleviate the over-harvesting of wild resources. However, the accumulation of intermediate secondary metabolites produced by plant cells in the culture medium will inhibit the further synthesis of the products. The continuous perfusion culture of plant cells can continuously bring the substances released by the cells into the medium to the outside of the reactor, effectively solving the problems of cell browning and product feedback inhibition. Compared with batch culture, c...

Claims

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Application Information

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IPC IPC(8): C12M3/00C12M1/04
CPCC12M29/04C12M29/08
Inventor 唐宁陈全战张边江
Owner NANJING XIAOZHUANG UNIV
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