Bioreactor and application thereof

Abstract

The invention relates to a bioreactor and an application thereof. The bioreactor comprises a main body part, wherein the main body part comprises a first cone, a cylinder and a second cone; a first cavity is formed in the first cone; a second cavity is formed in the cylinder, and the height-diameter ratio of the cylinder is 3-8; a third cavity is formed in the second cone, a ventilation unit is arranged in the third cavity, and the ventilation unit comprises a plate-shaped main body and a ventilation hole; and the first cavity, the second cavity and the third cavity are communicated. The bioreactor is wide in operable range of cell culture working volume, overcomes the defect that the operable range of the working volume of a shake flask, a shake bag reactor and a stirring type reactor islimited, can replace multi-step shake flask culture and shake bag reactor culture in the expanding culture process, reduces the number of times of transferring and expanding culture, and reduces the pollution risk; and the oxygen transfer efficiency is high, the problem of low oxygen transfer efficiency of a shake flask and a shake bag reactor is solved, high-density cell culture can be supported,open type operation is avoided, and the bacterial contamination risk is low.

Description

technical field

[0001] The invention relates to the technical field of cell expansion and culture, in particular to a bioreactor and its application. Background technique

[0002] In the past three decades, with the advancement of genetic engineering and cell culture technology, mammalian cell culture has been widely used in the biopharmaceutical industry.

[0003] Animal cell culture can be divided into adherent culture and suspension culture. Adherent culture systems mainly include T-flasks, spinner bottles, hollow fibers, microcarriers, flat-plate bioreactors and other culture systems. Due to the small surface area provided by the unit volume of adherent culture for cell growth, and the poor mass transfer effect and uniformity of the culture medium, it is not conducive to expanding the culture. Suspension culture mainly includes culture systems such as shake flasks, shaking bags, bubbling reactors, airlift reactors, and stirred reactors. Stirred bioreactors are equippe...

Examples

Embodiment 1

[0064] combined with figure 1 And attached figure 2 , the present embodiment provides a bioreactor, including a main body 101, the main body 101 includes a first cone 102, a cylinder 103 and a second cone 104, the first cone 102, cylinder 103 and second The cones 104 are sequentially connected along the height direction of the main body 101, the first cone 102 is located at the top of the main body 101, the second cone 104 is located at the bottom of the main body 101, and the first cone 102 and the second cone 102 are located at the top of the main body 101. The tips of the two cones 104 all face away from the cylinder 103; wherein:

[0065] The inside of the first cone 102 forms a first cavity 105, and the top of the first cone 102 is provided with an air outlet 106;

[0066] A second cavity 107 is formed inside the cylinder 103, and the aspect ratio of the cylinder 103 is between 3-8;

[0067] The inside of the second cone 104 forms a third cavity 108, the bottom of the...

Embodiment 2

[0087] This example provides a method for cell expansion using the bioreactor described in Example 1.

[0088] Specific steps are as follows:

[0089] (1) Take a Working Cell Bank (WCB) of suspended mammalian cells, thaw and recover in a 37°C water bath, and culture it in a 125mL or 250mL shake flask for 2-5 days. The culture conditions are as follows:

[0090] Temperature: 36-37°C, speed 100-150rpm, CO 2 Concentration 5-7%;

[0091] The selected medium can be selected by those skilled in the art as a commercial medium suitable for the cells.

[0092] (2) Take 20-50 mL of the cell suspension obtained from the culture in step (1) and introduce it into the bioreactor through the feed / outlet 115, and add 5-10 times the volume of the cell suspension as described in step (1) medium, so that the volume of the cell suspension in the bioreactor reaches 150-300mL, cultured at 36-37°C, and fed with 5-7% CO 2 Mixed gas with air, where the ventilation rate is 0.4vvm.

[0093] (3) Aft...

Embodiment 3

[0097] This embodiment provides a method for cell expansion using the bioreactor described in Embodiment 1. Compared with Embodiment 2, the difference lies in:

[0098] The suspension mammalian cells used in step (1) are CHO cells;

[0099] The medium used in step (1) is CD CHO medium.

technical field

[0001] The invention relates to the technical field of cell expansion and culture, in particular to a bioreactor and its application. Background technique

[0002] In the past three decades, with the advancement of genetic engineering and cell culture technology, mammalian cell culture has been widely used in the biopharmaceutical industry.

[0003] Animal cell culture can be divided into adherent culture and suspension culture. Adherent culture systems mainly include T-flasks, spinner bottles, hollow fibers, microcarriers, flat-plate bioreactors and other culture systems. Due to the small surface area provided by the unit volume of adherent culture for cell growth, and the poor mass transfer effect and uniformity of the culture medium, it is not conducive to expanding the culture. Suspension culture mainly includes culture systems such as shake flasks, shaking bags, bubbling reactors, airlift reactors, and stirred reactors. Stirred bioreactors are equippe...

Examples

Embodiment 1

[0064] combined with figure 1 And attached figure 2 , the present embodiment provides a bioreactor, including a main body 101, the main body 101 includes a first cone 102, a cylinder 103 and a second cone 104, the first cone 102, cylinder 103 and second The cones 104 are sequentially connected along the height direction of the main body 101, the first cone 102 is located at the top of the main body 101, the second cone 104 is located at the bottom of the main body 101, and the first cone 102 and the second cone 102 are located at the top of the main body 101. The tips of the two cones 104 all face away from the cylinder 103; wherein:

[0065] The inside of the first cone 102 forms a first cavity 105, and the top of the first cone 102 is provided with an air outlet 106;

[0066] A second cavity 107 is formed inside the cylinder 103, and the aspect ratio of the cylinder 103 is between 3-8;

[0067] The inside of the second cone 104 forms a third cavity 108, the bottom of the...

Embodiment 2

[0087] This example provides a method for cell expansion using the bioreactor described in Example 1.

[0088] Specific steps are as follows:

[0089] (1) Take a Working Cell Bank (WCB) of suspended mammalian cells, thaw and recover in a 37°C water bath, and culture it in a 125mL or 250mL shake flask for 2-5 days. The culture conditions are as follows:

[0090] Temperature: 36-37°C, speed 100-150rpm, CO 2 Concentration 5-7%;

[0091] The selected medium can be selected by those skilled in the art as a commercial medium suitable for the cells.

[0092] (2) Take 20-50 mL of the cell suspension obtained from the culture in step (1) and introduce it into the bioreactor through the feed / outlet 115, and add 5-10 times the volume of the cell suspension as described in step (1) medium, so that the volume of the cell suspension in the bioreactor reaches 150-300mL, cultured at 36-37°C, and fed with 5-7% CO 2 Mixed gas with air, where the ventilation rate is 0.4vvm.

[0093] (3) Aft...

Embodiment 3

[0097] This embodiment provides a method for cell expansion using the bioreactor described in Embodiment 1. Compared with Embodiment 2, the difference lies in:

[0098] The suspension mammalian cells used in step (1) are CHO cells;

[0099] The medium used in step (1) is CD CHO medium.

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