CN208038461U - Cell culture reactor - Google Patents

Abstract

The utility model discloses a kind of cell culture reactors.The cell culture reactor includes tank body, air inlet pipe and probe tube.Tank body has reaction chamber and is communicated in the air inlet of reaction chamber, gas outlet, adding mouth and sample tap, the size of reaction chamber is gradually reduced by bottom to the top of reaction chamber, the bottom surface of reaction chamber has tapered protrusion, one end of air inlet pipe is located at the outside of tank body and the end with bioreactor for being connected to, the other end of air inlet pipe extends to the bottom of tapered protrusion by air inlet, one end of probe tube is located at the outside of tank body and the end with bioreactor for being connected to, the other end of probe tube extends to the bottom of tapered protrusion by sample tap, gas outlet and adding mouth are connected to bioreactor respectively.The cell culture reactor oxygen delivery capacity is high, oxygen is evenly distributed, reduces cellular damage and improves cell culture efficiency and production cost is low.

Description

Cell culture reactor

Technical field

The utility model is related to a kind of cell culture reactors.

Background technology

Cell culture reactor for animal cell culture is broadly divided into two major classes, i.e. stirring-type cell culture reactor And non-stirred formula cell culture reactor, difference lies in whether agitating paddle is arranged inside cell culture reactor.Stirring-type Disadvantage of the reactor for animal cell culture is that shearing force is big, is easy to damage cell, although by various improvement, it should Problem there are still.In contrast, the shearing force that non-stirred formula cell culture reactor generates is small, the advantage in animal cell culture It is prominent.Non-stirred formula cell culture reactor generally makes cell culture medium by modes such as shake, rolling, rotation, swing, concussions It is moved in reactor, to keep cell suspended state and realize efficient gas exchanges.The cell culture of mainstream is anti-at present It is typical column mostly to answer the shape of device culture tank, is supplied oxygen for cell by way of air or oxygen injection.However, oxygen Gas can be constantly consumed after being ejected into culture pot bottom in uphill process, cause to occur oxygen concentration from top to bottom to successively decrease The case where, in order to ensure oxygen supply amount in culture supernatants, often sprayed using slightly excessive air or oxygen, however, mistake The air injection of amount, which often generates, foam and makes bubbles burst, same excessive oxygen spray also Cytotoxic.On the other hand, traditional Non-stirred formula cell culture reactor is difficult to generate higher media surface area, same shadow during exercise due to structure problem The absorption efficiency of oxygen is rung.Since such reactor is in the defect of oxygen supply respect, more complex control tower and phase are commonly used The dissolved oxygen probe of pass monitors the injection of air or pure oxygen, both makes complicated for operationization, while also improving cell culture and optimization Cost.

Utility model content

Based on this, it is necessary to provide that a kind of oxygen delivery capacity is high, oxygen is evenly distributed, reduces cellular damage and improves cell training Efficiency is supported to reduce the cell culture reactor of production cost.

A kind of cell culture reactor, including tank body, air inlet pipe and probe tube, the tank body have reaction chamber and company Air inlet, gas outlet, adding mouth and the sample tap of the reaction chamber are passed through, the size of the reaction chamber is by the reaction chamber Bottom to top is gradually reduced, and the bottom surface of the reaction chamber has tapered protrusion, and one end of the air inlet pipe is located at the tank body Outside and the end for being connected to bioreactor, the other end of the air inlet pipe extends to the cone by the air inlet The bottom of shape protrusion, one end of the probe tube are located at the outside of the tank body and the end and are used to connect with the bioreactor Logical, the other end of the probe tube extends to the bottom of the tapered protrusion, the gas outlet and institute by the sample tap Adding mouth is stated to be connected to the bioreactor respectively.

In one of the embodiments, further include sample-adding pipe, it is described sample-adding pipe one end be located at the tank body outside and For being connected to bioreactor, the other end of the sample-adding pipe is extended close to by the adding mouth in the taper at the end The top of protrusion.

Further include escape pipe in one of the embodiments, one end of the escape pipe be located at the outside of the tank body and For being connected to the bioreactor, it is convex that the other end of the escape pipe by the sample tap extends to the taper at the end The bottom risen.

Further include the first sealing-plug in one of the embodiments, the air inlet and the gas outlet, which merge, to be shared Opening, the air inlet and the gas outlet merge to form air inlet/outlet, and first sealing-plug seals the air inlet/outlet, The escape pipe and the air inlet pipe are arranged in first sealing-plug.

The adding mouth is located at the top of the tank body in one of the embodiments, the sample tap and it is described into Gas outlet is respectively positioned on the tank wall and the position of the sample tap and the air inlet/outlet is opposite.

Further include the second sealing-plug in one of the embodiments, second sealing-plug seals the adding mouth, described Sample-adding pipe is arranged in second sealing-plug.

Further include third sealing-plug in one of the embodiments, the third sealing-plug seals the sample tap, described Probe tube is arranged in the third sealing-plug.

The bottom surface of the reaction chamber is rounded in one of the embodiments, the bottom surface radius of the tapered protrusion and institute The bottom surface radius for stating reaction chamber is equal.

The air inlet pipe is close to the inner wall of the reaction chamber and extends to the tapered protrusion in one of the embodiments, Bottom, the inner wall that the probe tube is close to the reaction chamber extend to the bottom of the tapered protrusion.

The tapered protrusion is round bottom polygonal pyramid shape protrusion in one of the embodiments,.

There is tapered protrusion, air inlet pipe to be communicated in air inlet simultaneously for the bottom surface of above-mentioned cell culture reactor, institute's reaction chamber The bottom of tapered protrusion is extended to, probe tube is communicated in sample tap and extends to the bottom of tapered protrusion, by by oxygen spray It is the cell oxygen supply in culture solution to the bottom of reaction chamber, in this process, oxygen is gradually by the culture solution of reaction chamber bottom Cell utilize, oxygen content continuously decreases oxygen in uphill process in culture solution, due to reaction chamber size by reaction chamber bottom Portion to top is gradually reduced, and the size of reaction chamber is continuously decreased with the raising of height, to make culture solution in unit volume Oxygen content be held essentially constant, oxygen content is more uniformly spread, reduce cellular damage simultaneously improve cell culture efficiency.On The cell culture reactor stated, oxygen delivery capacity is high, and oxygen is evenly distributed, and production cost is minimized.

The bottom surface of above-mentioned cell culture reactor, reaction chamber has round bottom polygonal pyramid shape protrusion, is carrying out rotation culture When cell, culture solution and the pyramid of reaction chamber bottom collide, and to make culture solution form higher wave, are handed over to improve gas Transducing power, further increases cell culture efficiency.

Above-mentioned cell culture reactor, the round bottom polygonal pyramid shape protrusion being had based on reaction chamber bottom surface and reaction chamber The structure that size is gradually reduced by bottom to the top of reaction chamber, can reach lower concentration oxygen supply, higher efficiency it is thin Born of the same parents cultivate, and bubbles volume of the excessive oxygen supply to murder by poisoning and the generation of cell are reduced, to reduce toxigenic capacity and raising Culture efficiency.

Description of the drawings

Fig. 1 is the cell culture reactor schematic diagram described in an embodiment.

Reference sign

10, cell culture reactor；100, tank body；110, reaction chamber；120, it is loaded neck；130, neck is sampled；140, Gas port neck；150, tapered protrusion；200, air inlet pipe；300, probe tube；400, sample-adding pipe；500, escape pipe；600, first is close Blocking；700, the second sealing-plug；800, third sealing-plug.

Specific implementation mode

The utility model is more fully retouched below with reference to relevant drawings for the ease of understanding the utility model, It states.The preferred embodiment of the utility model is given in attached drawing.But the utility model can in many different forms come in fact It is existing, however it is not limited to embodiment described herein.Make public affairs to the utility model on the contrary, purpose of providing these embodiments is The understanding for opening content is more thorough and comprehensive.

It should be noted that when element is referred to as " being fixed on " another element, it can be directly on another element Or there may also be elements placed in the middle.When an element is considered as " connection " another element, it can be directly connected to To another element or it may be simultaneously present centering elements.

Unless otherwise defined, all of technologies and scientific terms used here by the article is led with the technology for belonging to the utility model The normally understood meaning of technical staff in domain is identical.Terminology used in the description of the utility model herein only be The purpose of description specific embodiment, it is not intended that in limitation the utility model.Term as used herein "and/or" includes Any and all combinations of one or more relevant Listed Items.

Shown in Figure 1, the present embodiment relates to a kind of cell culture reactor 10.The cell culture reactor 10 includes Tank body 100, air inlet pipe 200 and probe tube 300.Preferably, probe tube 300 can be plastic flexible pipe.Tank body 100 can be steel Prepared material is made.

Tank body 100 has reaction chamber 110 and is communicated in air inlet, gas outlet, adding mouth and the sampling of reaction chamber 110 Mouthful.The size of reaction chamber 110 is gradually reduced by bottom to the top of reaction chamber 110.The bottom surface of reaction chamber 110 has tapered protrusion 150.Whole tank body 100 is in positive horizontalization cone structure.It should be noted that tapered protrusion 150 is that raised top is tapered Structure can be polygonal pyramid structure or conical structure, polygonal pyramid structure such as trigone wimble structure, tetragonous wimble structure etc..

Shown in Figure 1, one end of air inlet pipe 200 is located at the outside of tank body 100 and the end and is used to connect with bioreactor Logical, the other end of air inlet pipe 200 extends to the bottom of tapered protrusion 150 by air inlet.

One end of probe tube 300 is located at the outside of tank body 100 and the end with bioreactor for being connected to, probe tube 300 The other end bottom of tapered protrusion 150 is extended to by sample tap.

Gas outlet and adding mouth are connected to bioreactor respectively.

Shown in Figure 1, in one embodiment, above-mentioned cell culture reactor 10 further includes sample-adding pipe 400.Sample-adding One end of pipe 400 is located at the outside of tank body 100 and the end with bioreactor for being connected to, and the other end of sample-adding pipe 400 passes through Adding mouth is extended close in the top of tapered protrusion 150.Preferably, sample-adding pipe 400 can be plastic flexible pipe.

In one embodiment, above-mentioned cell culture reactor 10 further includes escape pipe 500.One end of escape pipe 500 Positioned at the outside of tank body 100 and the end with bioreactor for being connected to, and the other end of escape pipe 500 is extended to by sample tap The bottom of tapered protrusion 150.Preferably, escape pipe 500 can be plastic flexible pipe.

In one embodiment, above-mentioned cell culture reactor 10 further includes the first sealing-plug 600.Air inlet, outlet Mouth merges shared opening namely air inlet/outlet is collectively formed in air inlet, gas outlet merging.First sealing-plug 600 is sealed into outlet Mouthful, it is preferable that 600 detachable-type sealed air inlet/outlet of the first sealing-plug.Escape pipe 500 and air inlet pipe 200 are arranged in One sealing-plug 600.Escape pipe 500 and air inlet pipe 200, which are sealed with the first sealing-plug 600, to be coordinated.Preferably, air inlet, go out Gas port is in a diameter of 60mm of air inlet/outlet after integral structure.A diameter of 60mm of sample tap.Adding mouth it is a diameter of 100mm。

Shown in Figure 1, in one embodiment, above-mentioned cell culture reactor 10 further includes the second sealing-plug 700, Second sealing-plug 700 seals adding mouth, it is preferable that 700 detachable-type sealed adding mouth of the second sealing-plug.Sample-adding pipe 400 is worn In the second sealing-plug 700, it is loaded between pipe 400 and the second sealing-plug 700 and seals cooperation.

In one embodiment, above-mentioned cell culture reactor 10 further includes third sealing-plug 800, third sealing-plug 800 sealing sample taps, it is preferable that 800 detachable-type sealed sample tap of third sealing-plug.Probe tube 300 is arranged in third sealing Plug 800.Probe tube 300 coordinates with the sealing of third sealing-plug 800.

In one embodiment, the bottom surface of reaction chamber 110 is rounded, bottom surface radius and the reaction chamber 110 of tapered protrusion 150 Bottom surface radius it is equal.

Preferably, the bottom surface radius of reaction chamber 110 is 200mm, and the bottom surface radius of tapered protrusion 150 is also 200mm.Taper The height of protrusion 150 is 100mm.

In one embodiment, adding mouth is located at the top of tank body 100.Preferably, the edge of adding mouth extends outwardly and has The sample-adding neck 120 of 20mm-50mm.

Sample tap and air inlet/outlet are respectively positioned on 100 side wall of tank body and the position of sample tap and air inlet/outlet is opposite. Preferably, the edge of sample tap, which extends outwardly, the sampling neck 130 of 30mm-70mm.Preferably, the edge of air inlet/outlet is outwardly It is extended with the gas port neck 140 of 30mm-70mm.Preferably, the distance between sample tap and air inlet/outlet are 300mm.

Further, bottom 300mm of the air inlet/outlet apart from tank body 100.Bottom 300mm of the sample tap apart from tank body 100.

Shown in Figure 1, in one embodiment, the inner wall that air inlet pipe 200 is close to reaction chamber 110 extends to tapered protrusion 150 bottom, the inner wall that probe tube 300 is close to reaction chamber 110 extend to the bottom of tapered protrusion 150.

In one embodiment, tapered protrusion 150 is round bottom polygonal pyramid shape protrusion 150, for example, round bottom triangular pyramid protrusion 150, round bottom pyramid shape protrusion 150, round bottom pentagonal pyramid shape protrusion 150 etc..Above-mentioned cell culture reactor 10, based on reaction The size of round bottom polygonal pyramid shape protrusion 150 and reaction chamber 110 that 110 bottom surface of chamber has by reaction chamber 110 bottom to top The structure being gradually reduced, can reach the oxygen supply of lower concentration, and the cell culture of higher efficiency reduces excessive oxygen supply pair The bubbles volume of murder by poisoning and the generation of cell, to reduce toxigenic capacity and improve culture efficiency.Above-mentioned cell culture is anti- Answer device 10, the bottom surface of reaction chamber 110 that there is round bottom polygonal pyramid shape protrusion 150, when carrying out rotation culture cell, reaction chamber 110 The culture solution of bottom is collided with pyramid, to make culture solution form higher wave, to improve gas exchange capacity, further Ground improves cell culture efficiency.

Preferably, the bottom surface of tank body 100 is recessed upwards, to save material, lowers being produced into for cell culture reactor 10 This.

Drosophila S 2 cells are cultivated using above-mentioned cell culture reactor 10：

Cell culture reactor 10 after sterilizing is placed on specific ring rail rocking bioreactor, sample-adding is managed 400, air inlet pipe 200, escape pipe 500 and probe tube 300 orifice corresponding with bioreactor respectively.It is managed by being loaded 400 are added the Sf900II SFM culture mediums that 15L is used for Insect cellculture, and 1L drosophilas S2 then is added by being loaded pipe 400 again Cell culture fluid, density are 20 × 10⁶A S2 cells/mL.Carry out cell culture in such a way that ring rail rocks, during which by into Tracheae 200 is passed through filtered air, the gas after being exchanged by the discharge of escape pipe 500.In culture, cell culture fluid can be with Bottom pyramidal protrusion 150 is collided, and to form higher wave, improves the surface area of cell culture fluid, is handed over to improve gas Transducing power.1mL cell culture fluids are taken out from probe tube 300 daily during culture, for detecting cell density, at the 5th day, carefully Born of the same parents' density can reach 30 × 10⁶A S2 cells/mL simultaneously enters plateau, and cell density starts to decay after the 11st day.And it uses and passes The bioreactor culture tank culture S2 cells of system, at the 5th day, general cell density only had 18-20 × 10⁶A cell/mL, Entered plateau to the 7th to 8 day, cell density starts to decay after 12 to 13 days.As it can be seen that being reacted using above-mentioned cell culture Device 10 cultivates Drosophila S 2 cells can faster reach plateau namely cell culture reactor compared to traditional bioreactor The efficiency of 10 culture cells greatly improves.

Chinese hamster ovary celI is cultivated using above-mentioned cell culture reactor 10：

Cell culture reactor 10 after sterilizing is placed on specific ring rail rocking bioreactor, sample-adding is managed 400, air inlet pipe 200, escape pipe 500 and probe tube 300 orifice corresponding with bioreactor respectively.It is managed by being loaded 400 are added the commercialization culture medium ProCHO5 that 12L is used for Chinese hamster ovary celI culture, then thin by being loaded the addition of pipe 400 3LCHO again Born of the same parents' culture solution, density 5x10⁶A Chinese hamster ovary celI/mL.The cell is the CHO stabilization cells for expressing hepatocyte growth factor (HGF) Strain.Cell culture is carried out in such a way that ring rail rocks, filtered air is during which passed through by air inlet pipe 200, passes through outlet Gas after the discharge exchange of pipe 500.In culture, cell culture fluid can with raised 150 local collisions of bottom pyramidal, to be formed Higher wave improves the surface area of cell culture fluid, to improve gas exchange capacity.After culture 7 days, from probe tube 300 take out all cell culture fluids, are purified, and HGF reagents are can get.And use traditional bioreactor culture tank culture The CHO stable cell lines of hepatocyte growth factor (HGF) are expressed, cell density growth is slower, the time required to reaching plateau Later, general technique is that harvest cell culture fluid is purified after cultivating 9-10 days.As it can be seen that anti-using above-mentioned cell culture The required density of purifying namely cell can faster be reached compared to traditional bioreactor by answering device 10 to cultivate Chinese hamster ovary celI The efficiency that cultivation reactor 10 cultivates cell greatly improves.

There is tapered protrusion 150, air inlet pipe 200 to be connected to for the bottom surface of above-mentioned cell culture reactor 10, institute's reaction chamber 110 In air inlet and the bottom of tapered protrusion 150 is extended to, probe tube 300 is communicated in sample tap and extends to tapered protrusion 150 Bottom, by being that the cell in culture solution is supplied oxygen by oxygen spray to the bottom of reaction chamber 110, in this process, oxygen is gradual It being utilized by the cell in 110 bottom culture solution of reaction chamber, oxygen content continuously decreases oxygen in uphill process in culture solution, due to The size of reaction chamber 110 is gradually reduced by bottom to the top of reaction chamber 110, the size of reaction chamber 110 with height raising And continuously decrease, to make the oxygen content of culture solution in unit volume be held essentially constant, oxygen content is more uniformly spread, It reduces cellular damage and improves cell culture efficiency.Above-mentioned cell culture reactor 10, oxygen delivery capacity is high, and oxygen distribution is equal Even, production cost is minimized.

Each technical characteristic of embodiment described above can be combined arbitrarily, to keep description succinct, not to above-mentioned reality It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited In contradiction, it is all considered to be the range of this specification record.

Above-described embodiments merely represent several embodiments of the utility model, the description thereof is more specific and detailed, But it should not be understood as limiting the scope of the patent of the utility model.It should be pointed out that for the common of this field For technical staff, without departing from the concept of the premise utility, various modifications and improvements can be made, these all belong to In the scope of protection of the utility model.Therefore, the protection domain of the utility model patent should be determined by the appended claims.

Claims (10)

1. a kind of cell culture reactor, which is characterized in that including tank body, air inlet pipe and probe tube, the tank body has anti- It answers chamber and is communicated in air inlet, gas outlet, adding mouth and the sample tap of the reaction chamber, the size of the reaction chamber is by institute Bottom to the top for stating reaction chamber is gradually reduced, and the bottom surface of the reaction chamber has tapered protrusion, one end position of the air inlet pipe Be used to be connected to bioreactor in the outside of the tank body and the end, the other end of the air inlet pipe by the air inlet simultaneously The bottom of the tapered protrusion is extended to, one end of the probe tube is located at the outside of the tank body and the end is used for and the life Object reactor is connected to, and the other end of the probe tube extends to the bottom of the tapered protrusion by the sample tap, it is described go out Gas port and the adding mouth are connected to the bioreactor respectively.

2. cell culture reactor according to claim 1, which is characterized in that further include sample-adding pipe, the sample-adding pipe One end is located at the outside of the tank body and the end with bioreactor for being connected to, and the other end of the sample-adding pipe is added by described Sample mouth is extended close in the top of the tapered protrusion.

3. cell culture reactor according to claim 1, which is characterized in that further include escape pipe, the escape pipe One end is located at the outside of the tank body and the end with the bioreactor for being connected to, and the other end of the escape pipe passes through institute State the bottom that sample tap extends to the tapered protrusion.

4. cell culture reactor according to claim 3, which is characterized in that further include the first sealing-plug, the air inlet Mouthful and the gas outlet merge and share opening, the air inlet and the gas outlet merge to form air inlet/outlet, described the One sealing-plug seals the air inlet/outlet, and the escape pipe and the air inlet pipe are arranged in first sealing-plug.

5. cell culture reactor according to claim 4, which is characterized in that the adding mouth is located at the top of the tank body Portion, the sample tap and the air inlet/outlet are respectively positioned on the tank wall and the sample tap and the air inlet/outlet Position it is opposite.

6. according to the cell culture reactor described in claim 2-5 any one, which is characterized in that further include the second sealing Plug, second sealing-plug seal the adding mouth, and the sample-adding pipe is arranged in second sealing-plug.

7. according to the cell culture reactor described in claim 1-5 any one, which is characterized in that further include third sealing Plug, the third sealing-plug seal the sample tap, and the probe tube is arranged in the third sealing-plug.

8. according to the cell culture reactor described in claim 1-5 any one, which is characterized in that the bottom surface of the reaction chamber Rounded, the bottom surface radius of the tapered protrusion is equal with the bottom surface radius of the reaction chamber.

9. according to the cell culture reactor described in claim 1-5 any one, which is characterized in that the air inlet pipe is close to institute The inner wall for stating reaction chamber extends to the bottom of the tapered protrusion, and the inner wall that the probe tube is close to the reaction chamber extends to institute State the bottom of tapered protrusion.

10. according to the cell culture reactor described in claim 1-5 any one, which is characterized in that the tapered protrusion is Round bottom polygonal pyramid shape protrusion.