CN209128457U

CN209128457U - A kind of duct type attached cell culture domesticating device

Info

Publication number: CN209128457U
Application number: CN201821449254.1U
Authority: CN
Inventors: 刘鹏; 廖国阳
Original assignee: Institute of Medical Biology of CAMS and PUMC
Current assignee: Institute of Medical Biology of CAMS and PUMC
Priority date: 2018-09-05
Filing date: 2018-09-05
Publication date: 2019-07-19
Anticipated expiration: 2028-09-05

Abstract

The utility model discloses a kind of duct type attached cells to cultivate domesticating device, it is desirable to provide a kind of to put into low duct type attached cell culture domesticating device.It includes the first peristaltic pump, cell culture tube, gas tank, the first air filter, the second air filter, circulation barrel, aeration net, air pump and loading bucket；The cell culture tube is flat tube, is made of transparent material, and both ends setting, which is shunk, constitutes interconnecting piece；One end of the cell culture tube is connected with first pipe, and the other end is connected with second pipe；The other end of the first pipe protrudes into circulation barrel and close to the bottom of circulation barrel；The 8th valve is provided in the first pipe；In the first pipe, it is provided with branch pipe between cell culture tube and the 8th valve, the 5th valve, the 6th valve and the 7th valve are disposed on the branch pipe；One end to the other end, which is connect, on the second pipe, along second pipe with cell culture tube is disposed with the first valve and the first peristaltic pump.

Description

A kind of duct type attached cell culture domesticating device

Technical field

The utility model relates to animal cell culture equipment technical fields, train more particularly, to a kind of duct type attached cell Support domesticating device.

Background technique

From in the early 1950s, the technology of animal cell culture production virus comes out, to a large amount of cell culture production In nearly 100 year that product occurs, animal cell culture technology experienced tortuous development.And it is based on animal cell culture technology Mass cell culture bioreactors be often viral vaccine, protease, interferon, the production work of the biological products such as antibody Skill core.The bioreactor that industry mainly uses at present has the disadvantage that:

1 disposable bioreactor: by taking ten layer cell factory 3271 of Corning as an example.Cell factory will usually be placed on non- Cell is cultivated on the flat shelf of ordinary water.This closed environment of cell factory usually requires to be transferred in the Biohazard Safety Equipment of desinfection chamber It is operated, rinses in inoculating cell, PBS, nutrient solution, virus inoculation, harvest is perfused and rinses the multi-pass operations links such as residual Increase pollution risk, increases cost of labor.And culture medium reflux is easy blocking air filter film, can not regulate and control partial pressure.Simultaneously Cell factory is medical disposable material again, excessively high for industrial production cost.

2 microcarrier bioreactors: culture is not intuitive, and it is live can not to observe culture.Microcarrier usage amount increases cell Yield not will increase, and toxicity increases therewith instead.Microcarrier must just can guarantee that microcarrier is in suspension by being stirred continuously State, therefore cell shearing power is caused, influence cell yield.The purchase of equipment and maintenance cost are high simultaneously.

3 suspension cell bioreactors: culture is not intuitive, and it is live can not to observe culture.It is very harsh to cell requirement, have When need to attached cell carry out gene editing, for the biological products such as viral vaccine production safety can not ensure, and Increase the R&D cycle.Cell and culture solution separation are complicated simultaneously, change experiment condition relative difficult.

4 roll culture bottle: rolling culture bottle cleaning is cumbersome, carries out passage to cell and needing to change culture medium.These steps are not Only increase workload, also increases the risk of cell contamination.Culture volume and cell density ratio are excessively high, and culture medium cost is high It is high.

In conclusion if can be in the automatization level of biological reactor for cell culture, personnel equipment's cost, to cell Rigors reduce the operation for having pollution risk, optimize in dynamic monitoring aspect, will reduce the life of product significantly Cost is produced, the quality of product is improved.

Utility model content

The purpose of this utility model is intended to overcome the shortcomings of the prior art, provide one kind can reduce production cost and The high duct type attached cell of product quality cultivates domesticating device.

In order to solve the above-mentioned technical problem, the utility model is achieved through the following technical solutions:

A kind of duct type attached cell culture domesticating device, including the first peristaltic pump, cell culture tube, gas tank, first is empty Air filter, the second air filter, circulation barrel are aerated net, air pump and loading bucket；The cell culture tube is flat tube, is adopted It is made of transparent material, both ends setting, which is shunk, constitutes interconnecting piece；One end of the cell culture tube is connected with first pipe, separately One end is connected with second pipe；The other end of the first pipe protrudes into circulation barrel and close to the bottom of circulation barrel；Described The 8th valve is provided on one pipeline；It is provided with branch pipe in the first pipe, between cell culture tube and the 8th valve, The 5th valve, the 6th valve and the 7th valve are disposed on the branch pipe；On the second pipe, along second pipe and thin Born of the same parents' culture tube connects one end to the other end and is disposed with the first valve and the first peristaltic pump；Third is plugged on the circulation barrel Pipeline and the first air inlet pipe, the third pipeline are located at one end in circulation barrel close to the bottom of circulation barrel；The aeration net connects It is connected to one end that the first air inlet pipe is located in circulation barrel；One end that first air inlet pipe is located at outside circulation barrel is connect with gas tank； First air filter is installed in the first air inlet pipe；It is plugged with the 4th pipeline and the second air inlet pipe in the loading bucket, The third pipeline is located at one end in loading bucket close to the bottom of loading bucket；Second air inlet pipe is located at one outside loading bucket End is connect with air pump；Second air filter is installed in the second air inlet pipe；On the second pipe, close to cell culture One end of pipe is connected separately with the second valve, third valve and the 4th valve by four-way connection；The second pipe is far from thin One end of born of the same parents' culture tube, third pipeline are located at one end outside circulation barrel and the 4th pipeline is located at one end outside loading bucket by three Pass joint connection, and junction is disposed with the tenth valve, the 9th valve and the 11st valve.

It preferably, further include being connected on second pipe, taming cell components close to the gradient at the first valve；It should It includes first gradient loading bucket, the second gradient loading bucket, the second peristaltic pump, third peristaltic pump and mixing that gradient, which tames cell components, Device；The 5th pipeline and third air inlet pipe are plugged on the first gradient loading bucket, the 5th pipeline is located in first gradient Close to the bottom of first gradient loading bucket, the other end is connected to vortex mixer lower part for one end in sample bucket, the second peristaltic pump peace Loaded on the 5th pipeline；The 6th pipeline and the 4th air inlet pipe, the 6th pipeline position are plugged on the second gradient loading bucket In one end in the second gradient loading bucket close to the bottom of the second gradient loading bucket, the other end is connected to vortex mixer lower part, described Third peristaltic pump is installed on the 6th pipeline；The upper end of the vortex mixer is connected to second pipe.

It preferably, further include the ultraviolet lamp sterilized for cell culture tube.

Preferably, the cell culture tube is more, is sequentially connected in series by connecting hose.

Preferably, the air pump is pumped using vacuum film.

Preferably, the cell culture tube is made of transparent glass.

Compared with prior art, the utility model has the advantages that

(1) the utility model, which only has, needs the docking for carrying out hose in Biohazard Safety Equipment and splits work when replacing loading bucket Make, greatly reduces contamination probability.

(2) the utility model cost of parts is low, and can replace easily, and maintenance cost is cheap；Pipeline component makes to clean, disappear Poison, sterilizing are more convenient.

(3) the utility model is by the control of the flow velocity to peristaltic pump, can long-time gradient change cell culture environment, it is right In taming cell adapted serum free medium, the living environments such as different pH are of great significance.

(4) pipeline is not high to the requirement for horizontality of holding plane in the utility model, and construction cost is low.Compared to cell factory By careful horizontal positioned conveniently many again after the linker constitutional balance of side.

(5) the utility model can observe cell culture situation, cytopathy by microscope directly on cell culture pipeline Become CPE situation, and the composition transfer of the real-time sampling Detection culture solution of system outlet can be passed through.

(6) since attached cell is fixed in the duct, it is possible to which low cost changes the living environment such as temperature of cell at any time The parameters such as degree, pH, partial pressure, ammonia, osmotic pressure.In the entire production cycle, cell can enter different growth phases, institute The optimum condition needed has very big difference, is changed according to process requirement and is suitble to the parameter of cell growth phase for high yield to Guan Chong It wants.

(7) the utility model solves the personnel equipment's cost residence height for producing biological products based on attached cell at present Under not, semi-manufactured goods quality is unstable, and process detects many puzzlements such as inconvenience, can fully meet the extensive training of attached cell It supports and tames.

Detailed description of the invention

In order to illustrate the embodiment of the utility model or the technical proposal in the existing technology more clearly, below will be to embodiment Or attached drawing needed to be used in the description of the prior art is briefly described, it should be apparent that, the accompanying drawings in the following description is only It is some embodiments of the utility model, for those of ordinary skill in the art, in the premise not made the creative labor Under, it can also be obtained according to these attached drawings other attached drawings.

Fig. 1 is the structural schematic diagram of the utility model one embodiment.

Fig. 2 is the structural schematic diagram of cell culture tube in Fig. 1.

Fig. 3 is the sectional view of cell culture tube in Fig. 2.

Fig. 4 is the schematic diagram of more root cells culture tubes connection.

Fig. 5 is the schematic diagram that gradient tames cell components.

Specific embodiment

The following will be combined with the drawings in the embodiments of the present invention, carries out the technical scheme in the embodiment of the utility model Clearly and completely describe, it is clear that the described embodiments are only a part of the embodiments of the utility model, rather than whole Embodiment.Based on the embodiments of the present invention, those of ordinary skill in the art are under that premise of not paying creative labor Every other embodiment obtained, fall within the protection scope of the utility model.In the following description, in order to clearly demonstrate originally The structure and working method of utility model will be described on the basis of attached drawing by many Directional words, but should be incited somebody to action The Word Understandings such as "front", "rear", "left", "right", "upper", "lower" are not construed as word of limitation for convenience of term.

Embodiment 1

A kind of duct type attached cell culture domesticating device, as shown in Figure 1-3, include the first peristaltic pump 12, cell culture Pipe 13, gas tank 15, the first air filter 16, the second air filter 14, circulation barrel 17 are aerated net 18, air pump 21, Yi Jishang Sample bucket 22；It is characterized by: the cell culture tube 13 is flat tube, it is made of transparent material, both ends setting, which is shunk, to be constituted Interconnecting piece 19；One end of the cell culture tube 13 is connected with first pipe 23, and the other end is connected with second pipe 24；Described The other end of one pipeline 23 protrudes into circulation barrel 17 and close to the bottom of circulation barrel 17；Is provided in the first pipe 23 Eight valves 8；In the first pipe 23, branch pipe 34, the branch pipe are provided between cell culture tube 13 and the 8th valve 8 The 5th valve 5, the 6th valve 6 and the 7th valve 7 are disposed on 34；On the second pipe 24, along second pipe 24 with Cell culture tube 13 connects one end to the other end and is disposed with the first valve 1 and the first peristaltic pump 12；On the circulation barrel 17 It is plugged with third pipeline 25 and the first air inlet pipe 35, the third pipeline 25 is located at one end in circulation barrel 17 close to circulation barrel 17 Bottom；The aeration net 18 is connected to one end that the first air inlet pipe 35 is located in circulation barrel 17；First air inlet pipe 35 It is connect in one end outside circulation barrel 17 with gas tank 15；First air filter 16 is installed in the first air inlet pipe 35；It is described The 4th pipeline 31 and the second air inlet pipe 36 are plugged in loading bucket 22, the third pipeline 25 is located at the termination in loading bucket 22 The bottom of nearly loading bucket 22；One end that second air inlet pipe 36 is located at outside loading bucket 22 is connect with air pump 21；Described second is empty Air filter 14 is installed in the second air inlet pipe 36；Pass through four on the second pipe 24, close to one end of cell culture tube 13 Pass joint is connected separately with the second valve 2, third valve 3 and the 4th valve 4；Second valve 2, third valve 3 and the 4th valve 4 Gas or liquid that instillation process needs can be distinguished as needed；One end far from cell culture tube 13 of the second pipe 24, One end that third pipeline 25 is located at one end outside circulation barrel 17 and the 4th pipeline 31 is located at outside loading bucket 22 passes through three-way connection Connection, and junction is disposed with the tenth valve 10, the 9th valve 9 and the 11st valve 11.

It further includes the ultraviolet lamp 20 sterilized for cell culture tube 13, convenient for the disinfection of cell culture tube.The air pump 21 are preferably pumped using vacuum film, it is clear that other pump housings also can be used.The cell culture tube 13 preferably uses transparent glass system At, it is clear that transparent plastic also can be used.

Embodiment 2

A kind of duct type attached cell culture domesticating device, wherein the cell culture tube 13 is more, soft by connecting Pipe 39 is sequentially connected in series, as shown in figure 4, remaining is substantially the same manner as Example 1.

Embodiment 3

A kind of duct type attached cell cultivates domesticating device, further includes being connected to the in addition to implementing structure described in wound 1 Cell components are tamed on two pipelines 24, close to the gradient at the first valve 1, as shown in Figure 5；The gradient tames cell components First gradient loading bucket 27, the second gradient loading bucket 28, the second peristaltic pump 29, third peristaltic pump 30 and vortex mixer 26；Described The 5th pipeline 32 and third air inlet pipe 37 are plugged on one gradient loading bucket 27, the 5th pipeline 32 is located at first gradient loading Close to the bottom of first gradient loading bucket 27, the other end is connected to 26 lower part of vortex mixer for one end in bucket 27, and described second wriggles Pump 29 is installed on the 5th pipeline 32；It is plugged with the 6th pipeline 33 and the 4th air inlet pipe 38 on the second gradient loading bucket 28, 6th pipeline 33 is located at one end in the second gradient loading bucket 28 close to the bottom of the second gradient loading bucket 28, the other end with The connection of 26 lower part of vortex mixer, the third peristaltic pump 30 are installed on the 6th pipeline 33；The upper end and second of the vortex mixer 26 Pipeline 24 is connected to.

The application method of the utility model is as follows:

[inoculating cell]

The culture solution for digesting the cell to fall off and required volume is put into loading bucket 22 and is mixed, the first valve, the are opened Five valves, the 6th valve, the 7th valve, the tenth valve, the 11st valve close the 8th valve and the 9th valve.7th valve It flanks air filter outside and is exhausted and be placed in eminence.Air pump 21 is opened, liquid in loading bucket 22 is slowly pumped into cell culture tube In 13, until culture solution liquid level is at branch outlet.Air pump 21 and all valves, inducing cell patch are closed after loading Wall.Static adherent length of time is subject to microscope and is observed.Cell it is adherent completely after, start the cycle over culture cell manipulation.

[cell expansion culture]

According to high pressure sterilization after wanting widened ratio to prepare required amount of cell culture tube 13 and being connected with connection hose. The 8th valve and the 9th valve are closed, the first valve, the 5th valve, the 6th valve, the 7th valve, the tenth valve, the tenth are opened One valve opens air pump 21, and the digestive juice for preparing in advance in loading bucket 22 is pumped into system.Digestive juice is discharged in branch outlet. As the liquid level of loading bucket 22 reduces, pipeline starts to pump air inlet body, side by side the digestive juice in vacant duct.Close the 11st valve and Air pump 21 opens the 9th valve and 12 flow velocity of peristaltic pump is turned up, and with the nutrient solution flushing pipe of circulation barrel 17, receives in branch outlet Collect cast-off cells.The cell culture tube 13 connected is connected in system after collection, according to being connect in specific embodiment The operating procedure of kind cell completes the again adherent of cell.

[Cyclic culture cell]

Air pump 21 is opened, the pre-assigned nutrient solution of 17 certain volume of circulation barrel is pumped by loading bucket 22.Gas tank 15 is pressed into Mixed gas simultaneously controls gas flow foam is avoided to generate.The 6th valve of the 5th valve, the 7th valve, the 11st valve are closed, Open peristaltic pump 12 and the first valve, the 8th valve, the 9th valve and the tenth valve.It starts the cycle over.Nutrient solution is through peristaltic pump 12 It is pumped into second pipe, is back to circulation barrel 17 after pipeline slowly flows across the cell culture tube 13 of inoculating cell.Certain time Afterwards, setting peristaltic pump 12 is reversely wriggled.

[process detection]

During system circulation, branch outlet is put into Biohazard Safety Equipment, successively opens the 7th valve, the 6th valve Sample is collected with the 5th valve.Successively closing the 5th valve, the 6th valve, the 7th valve terminates to receive sample.

[gradient domestication cell]

The good first gradient loading bucket 27 of recipe configuration and the second gradient loading bucket in gradient domestication component as required 28, gradient domestication component is connect at the first valve, the first peristaltic pump 29 and the second peristaltic pump 30 are set, make it at any time or body Long-pending growth and increase or decrease revolving speed.The 5th valve, the 6th valve, the 7th valve, the 9th valve are closed, the first valve is opened Door, the 8th valve open the first peristaltic pump, the second peristaltic pump and vortex mixer 26.Nutrient solution collection is carried out in circulation barrel 17.

[virus inoculation and harvest]

Configured viral solution is placed in loading bucket 22, opens the first valve, the 8th valve, the 11st valve, the tenth Valve.It opens air pump 21 and is pumped into air progress loading.Virus can automatic absorbing and infected cell.After completion of the sample, the tenth is closed One valve and air pump 21 are opened the 9th valve and peristaltic pump 12, are recycled.

After being incubated for a period of time, rate of circulating flow should be gradually increased with flushing pipe when entering the viral release stage, is finally closed Peristaltic pump 12 and the 9th valve are closed, the 11st valve and air pump 21 are opened.Air is pumped into from loading bucket 22, pushes second pipe Circulation barrel 17 is passed back into the virus harvest liquid in cell culture tube 13.Complete harvest.

If the mode of virus release is to sprout and do not lead to cell rupture death such as rabies viruses etc., one section can be incubated for After time, peristaltic pump 12 and the 9th valve are closed, opens the 11st valve and air pump 21.Air is pumped into reaction from loading bucket 22 Device pushes the virus harvest liquid in second pipe and cell culture tube 13 to pass back into circulation barrel 17.Only open the 9th valve the tenth One valve.Air pump 21 is opened to be evacuated.Harvest liquid is pumped into loading bucket 22, completes primary harvest.Then pass through new loading bucket 22 It is pumped into circulation barrel 17 and persistent collection is carried out with new nutrient solution.

The above is only the preferred embodiment of the utility model only, is not intended to limit the utility model, all at this Within the spirit and principle of utility model, any modification, equivalent replacement, improvement and so on should be included in the utility model Protection scope within.

Claims

1. a kind of duct type attached cell cultivates domesticating device, including the first peristaltic pump (12), cell culture tube (13), gas tank (15), the first air filter (16), the second air filter (14), circulation barrel (17) are aerated net (18), air pump (21), with And loading bucket (22)；It is characterized by: the cell culture tube (13) is flat tube, it is made of transparent material, both ends setting It shrinks and constitutes interconnecting piece (19)；One end of the cell culture tube (13) is connected with first pipe (23), and the other end is connected with Two pipelines (24)；The other end of the first pipe (23) protrudes into circulation barrel (17) and close to the bottom of circulation barrel (17)；Institute It states and is provided with the 8th valve (8) on first pipe (23)；On the first pipe (23), it is located at cell culture tube (13) and the 8th It is provided between valve (8) branch pipe (34), the 5th valve (5), the 6th valve (6) and is disposed on the branch pipe (34) Seven valves (7)；Connect on the second pipe (24), along second pipe (24) with cell culture tube (13) one end to the other end according to It is secondary to be provided with the first valve (1) and the first peristaltic pump (12)；Third pipeline (25) and first are plugged on the circulation barrel (17) Air inlet pipe (35), the third pipeline (25) are located at one end in circulation barrel (17) close to the bottom of circulation barrel (17)；The exposure Gas net (18) is connected to one end that the first air inlet pipe (35) is located in circulation barrel (17)；First air inlet pipe (35) is located at circulation The one end of bucket (17) outside is connect with gas tank (15)；First air filter (16) is installed on the first air inlet pipe (35)；Institute It states in loading bucket (22) and is plugged with the 4th pipeline (31) and the second air inlet pipe (36), the third pipeline (25) is located at loading bucket (22) bottom of one end in close to loading bucket (22)；Second air inlet pipe (36) is located at the outer one end of loading bucket (22) and gas Pump (21) connection；Second air filter (14) is installed on the second air inlet pipe (36)；On the second pipe (24), connect One end of nearly cell culture tube (13) is connected separately with the second valve (2), third valve (3) and the 4th valve by four-way connection (4)；One end of the second pipe (24) separate cell culture tube (13), third pipeline (25) are located at one of circulation barrel (17) outside End and the 4th pipeline (31) are located at the one end of loading bucket (22) outside by three-way connection connection, and junction is disposed with the Ten valves (10), the 9th valve (9) and the 11st valve (11).

2. duct type attached cell cultivates domesticating device according to claim 1, it is characterised in that: it further includes being connected to the Cell components are tamed on two pipelines (24), close to the gradient at the first valve (1)；It includes the first ladder that the gradient, which tames cell components, It spends loading bucket (27), the second gradient loading bucket (28), the second peristaltic pump (29), third peristaltic pump (30) and vortex mixer (26)；Institute It states and is plugged with the 5th pipeline (32) and third air inlet pipe (37) on first gradient loading bucket (27), the 5th pipeline (32) is located at One end in first gradient loading bucket (27) is close to the bottom of first gradient loading bucket (27), the other end and vortex mixer (26) lower part Connection, second peristaltic pump (29) are installed on the 5th pipeline (32)；Is plugged on the second gradient loading bucket (28) Six pipelines (33) and the 4th air inlet pipe (38), it is close that the 6th pipeline (33) is located at one end in the second gradient loading bucket (28) The bottom of second gradient loading bucket (28), the other end are connected to vortex mixer (26) lower part, and the third peristaltic pump (30) is installed on On 6th pipeline (33)；The upper end of the vortex mixer (26) is connected to second pipe (24).

3. duct type attached cell according to claim 1 or claim 2 cultivates domesticating device, it is characterised in that: it further includes being used for The ultraviolet lamp (20) of cell culture tube (13) disinfection.

4. duct type attached cell according to claim 1 or claim 2 cultivates domesticating device, it is characterised in that: the cell culture Managing (13) is more, is sequentially connected in series by connecting hose (39).

5. duct type attached cell according to claim 1 or claim 2 cultivates domesticating device, it is characterised in that: the air pump (21) It is pumped using vacuum film.

6. duct type attached cell according to claim 1 or claim 2 cultivates domesticating device, it is characterised in that: the cell culture Pipe (13) is made of transparent glass.