CN206014938U - Full-automatic cell continuous culture system - Google Patents

Abstract

This utility model provides a kind of full-automatic cell continuous culture system, at least includes：Control module, control platform, dark field microscope, cell culture table and the cell culture bags observed for cell culture online；Cell culture table includes cultivating bag bracket, rocking carrier and agitating device, and culture bag bracket is provided with curved plate and cell culture bags fixed component, and curved plate is provided with object lens insertion breach；Cell culture bags, including closed bag, are provided with transparent observing area in the middle part of its bottom surface；Bag is placed in cell culture table, and transparent observing area is consistent with object lens gap position；For the dark field microscope that cell culture is observed online, its object lens just can be inserted into the object lens insertion breach in cell culture table；Control platform connects dark field microscope and cell culture table respectively；Control module connects control platform.The continuous culture and observation that cell can be realized using this utility model, convenient and swift, substantially increase the efficiency of cultured cells.

Description

Full-automatic cell continuous culture system

Technical field

This utility model is related to a kind of full-automatic cell continuous culture system, is mainly used in culture and observation of cell.

Background technology

The immunocyte culture of human body therapy is presently used for, and mainly or Tissue Culture Dish is used under laboratory super-clean bench Or culture bottle is cultivated, and the density according to cell growth, continuous sub-bottle culture is carried out, this cell culture pattern, not only Take time and effort, simultaneously because needing different sub-bottles, sample-adding, be easier to pollute.At the same time, current use for laboratory is in sight The method for examining cell is also more original, includes sampling mostly and makes the link of specimen, operates relatively complicated, it is impossible to realize cell Continuous and Real Time Observation.

With the development and the progress of artificial intelligence of science and technology, the continuous culture of cell and observation are had begun to by people institute Concern, but also there is not a kind of continuous culture that can realize cell and the device of observation.

Utility model content

The shortcoming of prior art in view of the above, the purpose of this utility model are to provide a kind of full-automatic cell continuous Culture systems, it is possible to achieve the continuous culture of cell and observation, and the measure of the parameters to culture fluid.

For achieving the above object and other related purposes, this utility model provides a kind of full-automatic cell continuous training system System, the cell culture system at least include：Control module, control platform, the dark field for the online observation of cell culture show Micro mirror, cell culture table, cell culture bags and power module；

The cell culture table includes cultivating bag bracket, rocking carrier and agitating device, and the culture bag bracket is provided with arc Shape base plate and cell culture bags fixed component, the rocking carrier include swinging base, and the swinging base is provided with support arm, institute State culture bag bracket to be fixedly connected through support arm with the rocking carrier, the agitating device includes that pedestal, pedestal are provided with and wave Component, the swinging base are connected with component is waved, and the curved plate is provided with object lens insertion breach；

The cell culture bags, including closed bag, are provided with transparent observing area in the middle part of the bottom surface of the bag；The bag Body is placed in cell culture table, and the transparent observing area is consistent with object lens gap position；

The dark field microscope that is observed for cell culture online, including object lens, the object lens just can be inserted into thin Object lens insertion breach on born of the same parents' culture shaking table；

The control platform is connected to the cell culture dark field microscope of observation and cell culture table online respectively；

The control module connects control platform；

The power supply is powered for whole system.

For further optimizing design scheme, the dark field microscope that is observed for cell culture online includes illuminating Element, Moveable support, support base, photo-sensitive cell, caliber unit, A/D converting units, power module and control mould Block, the illumination component are arranged on the top of Moveable support, and the Moveable support is arranged in support base, described Caliber unit is connected to Moveable support bottom, and the object lens are arranged on caliber unit and corresponding under illumination component Side, the photo-sensitive cell are arranged on the light signal in caliber unit for gathering on object lens, the A/D converting units connection sense Optical element, the control module connect A/D converting units.

Preferably, the illumination component includes the lamp socket of circle, and is arranged on lamp socket and surrounds several concentric circulars row The center of circle of concentric circular described in the illuminating lamp of row and the home position of lamp socket are consistent.

Preferably, the lamp socket to object lens distance be 10-30 centimetre.

Preferably, the light source of the illuminating lamp forms dark field with object lens, meets the needs of dark field illumination.

Preferably, the illuminating lamp is LED.

Preferably, the support base includes motor, slide mechanism and base, the slide mechanism include slide block with Slide rail, the control module connect and control motor, the motor connection sliding block, and the slide block slides through slide rail and connects It is connected on base and horizontally moveable, the Moveable support is arranged on slide block.

For further optimizing design scheme, one or more accessory fixed hole positions are set on the curved plate, described Leave under accessory fixed hole position for installing or insert the space of outside appurtenances.

Preferably, the curved plate is provided with heating plate.

Preferably, the edge of the curved plate is provided with baffle plate.

Preferably, the cell culture bags fixed component is located at the both sides of curved plate, including lateral from curved plate two The platform that both sides extend, the platform are provided with fixed rod clamp and/or hold-down bars interposer.

Preferably, the swinging base is flat board, and two ends are provided with support arm, and culture bag bracket is not contacted with swinging base, Between leave space to install or insert outside appurtenances.

Preferably, the component that waves includes that hinged seat and at least one pumps mechanism, and the hinged seat sets On pedestal, the swinging base is hinged with hinged seat, and the mechanism that pumps is connected with swinging base or is mutually compressed.

For further optimizing design scheme, the cell culture bags include：The fixed component of bag both sides, the closing Formula upper face of bag body is provided with outside liquid adding mouth, outside liquid injection port, gas feed and gas outlet, the closed bag Bottom surface is provided with liquids recovery mouth and at least one sensor connector, the outside liquid adding mouth, outside liquid injection port, gas Aseptic snap joint is equipped with import, gas outlet and liquids recovery mouth.

Preferably, the closed bag surrounds a solid jointly by upper bag sheet, lower bag sheet, left side bag webs and right side bag webs The bottom of bag chamber, the left side bag webs and the right side bag webs is curved, and the left side bag webs and the right side bag webs are in Ellipse, the upper bag sheet and lower bag sheet rectangular.

Preferably, the closed bag innermost layer is polyethylene layer.

Preferably, the material used by the closed bag be five layers of composite construction, be followed successively by from the inside to the outside polyethylene layer, Adhesive layer, ethylene, propylene alcohol layer, adhesive layer and polyethylene layer.

Preferably, described sensor connector is provided with one or more.

Preferably, described sensor connector is noncontacting proximity sensor joint or touch sensor joint.

Preferably, the contactless sensor connector includes basal disc and sensor probe containing cavity, on the basal disc Transparent cut-off is provided with, the sensor probe containing cavity is connected with the basal disc and the side positioned at the transparent cut-off, described The opposite side of transparent cut-off is provided with conversion film.

Preferably, the film of changing causes color-changing membrane or dissolved oxygen to cause color-changing membrane into pH.

For further optimizing design scheme, the control platform includes switch board, and be arranged on switch board from Dynamic heating unit, preheating bag and peristaltic pump；The automatic heating unit is arranged and includes preheating table, and the preheating bag is arranged on preheating On plate, the peristaltic pump includes that the first peristaltic pump and the second peristaltic pump, the automatic heating unit also include cell culture flow container Body, the tank body preheat the inlet of bag by the first pipeline communication, and the outlet of the preheating bag is thin by second pipe connection Born of the same parents' culture bag, first peristaltic pump connect the first pipeline, and second peristaltic pump connects second pipe, the automatic heating list Unit, the first peristaltic pump and the second peristaltic pump link control module.

Preferably, gas mixing units are additionally provided with inside the switch board so that the gas outlet of the gas mixing units sets Put in the upper of switch board, and by the 3rd pipeline communication cell culture bags, the air inlet connection of the gas mixing units is outside Source of the gas, the gas mixing units link control module.

Preferably, pH value detection tabs and dissolved oxygen detection tabs are additionally provided with the switch board, and the pH value detection is visited Head and dissolved oxygen detection probe.

For further optimizing design scheme, in order to realize control of the control module to whole culture systems, the control Module includes：

Moveable support control unit, for controlling the movement of Moveable support；

Illumination component control unit, for controlling the switch of illumination component；

Unit is checked, for checking object lens are observed under current state content, and cell growth under current state The parameters of environment；

Storage element, for storing every ginseng of the content for the observing and record cell growth environment for needing to preserve Number；

Gas control unit, for controlling the gas in cell culture bags；

Cell culture fluid control unit, for controlling temperature and the supply of cell culture fluid.

Other side of the present utility model there are provided the dark field microscope for the online observation of cell culture Application：For observation of cell culture bag.

Other side of the present utility model there are provided cell culture bags application：For cultured cells.

As described above, full-automatic cell continuous culture system of the present utility model, has the advantages that：

(1) the continuous culture of cell is realized, while that heated, rocks, being loaded is carried out continuously；

(2) without the need for manually changing liquid, for the temperature of Cell sap can realize intelligent control, it is possible to achieve automatic liquid-feeding；

(3) user can be according to the status adjustment shaking table wobble frequency of cell culture and angle, it is ensured that cell is in optimal Growing environment；

(4) due to being provided with multiple fixed hole positions on curved plate in this utility model, it is thereby achieved that various spies Survey device probe to cell culture bags in various parameters detect；

(5) dark field microscope for being used for the online observation of cell culture is used for observation of cell sample, without the need for sampling, Ke Yishi The continuous Real Time Observation of existing cell；

(6) cell culture bags unique structure of the present utility model, can meet persistently sample-adding training under the conditions of a small amount of culture fluid Support, until it reaches culture demand；

(7) the 3D heterocyst culture bags of this utility model design, in cell cultivation process, the shake institute of cultivation platform Shearing force minimum is produced, the shearing force caused by cell is damaged also minimum；

(8) all joints of cell culture bags of the present utility model connect mode soon using aseptic, reduce training to greatest extent Support the risk of the cell contamination caused by operation；

(9) volume of cell culture bags of the present utility model can infinitely expand according to the amount of required culture；

(10) can be recorded and various parameters in real time inspection cell growth process using control system of the present utility model；

(11) the intelligent gas provided needed for cell culture.

Description of the drawings

Fig. 1 is shown as the schematic diagram of this utility model preferred embodiment cell culture table.

Fig. 2 is shown as this utility model and preferably cultivates bag bracket schematic diagram.

Fig. 3 is shown as another preferably culture bag bracket schematic diagram of this utility model.

Fig. 4 is shown as the schematic diagram of this utility model another preferred embodiment cell culture table.

Fig. 5 is shown as the schematic diagram of this utility model further embodiment cell culture table.

Fig. 6 is shown as being used for the dark field microscope structural representation that cell culture is observed online in this utility model.

Fig. 7 is shown as dark field illumination principle schematic in this utility model.

Fig. 8 is shown as the cell culture bags top schematic diagram of this utility model embodiment.

Fig. 9 is shown as the cell culture bags schematic bottom view of this utility model embodiment.

Figure 10 is shown as the cell culture bags Bag structure schematic diagram of this utility model preferred embodiment.

Dotted line index wire points to the bag that illustrated angle theory is blocked

Figure 11 is shown as the cell culture bags use state lower bag body top surface connecting line of this utility model preferred embodiment Schematic diagram.

Figure 12 is shown as the cell culture bags use state lower bag body bottom surface connecting line of this utility model preferred embodiment Schematic diagram.

Figure 13 is shown as the sensor connector profile of this utility model preferred embodiment.

Figure 14 is shown as control module structural framing figure.

Figure 15 is shown as whole system structural representation.

1 culture bag bracket

11 curved plates

111 accessory fixed hole positions

112 object lens insert breach

113 baffle plates

114 heating plates

12 cell culture bags fixed components

121 platforms

122 fix rod clamp

123 hold-down bars interposers

2 rocking carriers

21 swinging bases

211 support arms

3 agitating devices

31 wave component

311 hinged seats

312 pump mechanism

32 pedestals

4 cell culture bags

41 closed bags

411 outside liquid adding mouths

412 outside liquid injection ports

413 gas feeds

414 gas outlets

415 liquids recovery mouths

4161 upper bag sheets

4162 lower bag sheets

4163 left side bag webs

4164 right side bag webs

417 sensor connector

4171 are used for accessing the sensor connector that pH sensor is popped one's head in

4172 are used for accessing the sensor connector of dissolved oxygen (DO) sensor probe

41701 basal discs

41702 sensor probe containing cavities

41703 transparent cut-offs

418 aseptic snap joints

419 transparent observing areas

42 fixed components

431 silica gel tubes

432 female Luers

433 threeways

434 needleless samplers

435 air filters

436 air cutoff valves

437 folders

438 bands

44 seal areas

5 support bases

501 motors

502 slide blocks

503 bases

6 caliber units

61 object lens

7 control modules

8 lamp sockets

9 Moveable supports

10 photo-sensitive cells

13 switch boards

1301 first peristaltic pumps

1302 second peristaltic pumps

1303 PH detection tabs

1304 dissolved oxygen detection tabs

1305 gas outlets

1306 external cellular culture fluid

1307 second pipes

1308 first pipelines

1309 preheating bags

Specific embodiment

Embodiment of the present utility model is illustrated by particular specific embodiment below, those skilled in the art can be by this Content disclosed by description understands other advantages of the present utility model and effect easily.

Refer to Fig. 1 to Figure 15.It should be clear that structure, ratio, size depicted in this specification institute accompanying drawings etc., only uses To coordinate the content disclosed in description, so that those skilled in the art understands and reads, this practicality is not limited to new The enforceable qualificationss of type, therefore do not have technical essential meaning, the modification of any structure, the change of proportionate relationship or size Adjustment, under the effect for not affecting this utility model can be generated and the purpose that can be reached, still all should fall in institute of the present invention In the range of the technology contents of announcement are obtained and can be covered.Meanwhile, in this specification cited as " on ", D score, "left", "right", The term of " centre " and " one " etc., is merely convenient to understanding for narration, and is not used to limit enforceable scope of the invention, its phase Relation is altered or modified, under without essence change technology contents, is raised when enforceable model of the invention is also considered as.

Referring to Figure 15, the utility model discloses a kind of full-automatic cell continuous culture system, cell culture system is at least Including：Control module, control platform, the dark field microscope for the online observation of cell culture, cell culture table, cell training Foster bag and power module, control module connect control platform, and control platform connects and control cell culture table and use respectively In the dark field microscope that cell culture is observed online, the dark field microscope that is observed for cell culture online is thin for observing The cell that cultivates on born of the same parents' culture shaking table, the cell culture bags are arranged in cell culture table, and the power module is whole System power supply.

1 cell culture table of embodiment

Cell culture table as shown in Figure 1, including cultivating bag bracket 1, rocking carrier 2 and agitating device 3, the culture bag Bracket 1 is provided with curved plate 11 and cell culture bags fixed component 12, and the rocking carrier 2 includes swinging base 21, described waves Base 21 is provided with support arm 211, and the culture bag bracket 1 is fixedly connected through support arm 211 with the rocking carrier 2, described shakes Dynamic device 3 includes that pedestal 32, pedestal are provided with and wave component 31, and the swinging base 21 is connected with component 31 is waved.

In preferred embodiment as shown in Figure 2, the curved plate 11 is provided with object lens insertion breach 112.In order in cell Real Time Observation cell situation in continuous culture, is provided with transparent observing area in culture bag bottom, and object lens insertion breach 112 mates transparent Observation area, can be inserted into object lens, so as to utilize object lens, the direct cell situation from the observation of cell culture bag of bottom.

The curved plate 11 is used for the main part for housing arc 3D cell culture bags 4.The design of curved plate is caused In cell cultivation process, bottom can maintain arc to arc 3D cell culture bags, so as in cell cultivation process, cultivation platform Shake produced by shearing force minimum, shearing force that cell is caused is damaged also minimum.

Preferably, two side of the curved plate is arc.

Further, as shown in Fig. 2 one or more accessory fixed hole positions 111 can be arranged on the curved plate 11. Preferably, being provided with female thread preferably to fix outside appurtenances in the fixed hole position 111.Outside appurtenances can be various spies Survey the probe of device, such as pH value and oxygen dissolving value detection probe etc..Outside appurtenances can also be various outside liquid feed tubes, injection unit Part, drain pipe, extraneous gas delivery pipe etc. are easy to the accessory that cell is continuously cultivated.

The curved plate is the arc to lower recess in a state of use.Further, the object lens insert breach 112 It is arranged at the concave bottom region of curved plate.

Further, in preferred embodiment as shown in Figure 2, the edge of curved plate 11 is provided with baffle plate 113.Baffle plate can be prevented Cell culture bags are slipped from culture bag bracket, displacement.

In preferred embodiment as shown in Figure 3, the curved plate 11 is provided with heating plate 114.Heating plate can be trained in cell Heating when foster, to provide suitable temperature as cell culture.Control module can set the temperature needed for cell culture bags 4, The heating of control heating plate.

As shown in Fig. 2 the cell culture bags fixed component 12 may be provided in the both sides of curved plate, can be from arcuate bottom The platform 121 that plate both sides of the edge extend to both sides, which is provided with the part matched with the fixed component in cell culture bags.Example Can such as be the part fixed by mechanical force, viscous force, magnetic force etc., or screw and nut are set on platform 121 by carefully Born of the same parents' culture bag 4 is fixed on platform.In embodiment as shown in Figure 4,4 both sides of cell culture bags are provided with hold-down bars, now, Fixed rod clamp 122 and/or hold-down bars interposer 123 can be provided with platform, so as to realize fixing with cell culture bags 4 The detachable connection of rod.

In the rocking carrier 2, the design of support arm 211 can play a part of the high culture bag bracket 1 of support.Culture bag bracket 1 Accessory fixed hole position should leave for 111 times for installing or insert the space of outside appurtenances.In rocking carrier, support arm can be made A part and swinging base for swinging base 21 is integrally formed, can act also as a single part installed in swinging base it On, or can also be integrally formed with culture bag bracket 1.On the one hand support arm can be the company of cell culture bags and outside appurtenances Headspace is connect, on the other hand, due to the high culture bag bracket 1 of frame, it is only necessary to one is provided to swinging base 21 and small size is shaken Pendulum power, culture bag bracket 1 just can be in an interior swing in a big way.

In the preferred embodiments as shown in Fig. 1,4,5, swinging base 21 is flat board, and two ends are provided with support arm 211, culture bag Bracket 1 is not contacted with swinging base 21, between leave space to install or insert outside appurtenances, including various probes, object lens.

Described wave the mechanism that component 31 can be arbitrarily existing achievable rocking action.

In preferred implementation as shown in Figure 1, the component 31 that waves includes hinged seat 311 and about at least one Reciprocating mechanism 312, on pedestal 32, the swinging base 21 of rocking carrier 2 is hinged the hinged seat 311 with hinged seat 311, So as to provide a swing center, the mechanism 312 that pumps is connected with swinging base 21 or is mutually compressed, so that Pump mechanism 312 can promote swinging base 21 around swing center make reciprocating swing motion.

The hinged seat 311 can have in one or more, as shown in Figure 5 examples, at the middle part of pedestal 32, symmetrically set Two hinged seats 311 are put, so that waving more stable.Based on the work of the mechanism 312 that pumps, rocking carrier can be with Make yaw movements.

The mechanism 312 that pumps can be various existing achievable reciprocating mechanisms, for example electronic Push rod, linear electric motors, toggle etc..Specifically, for example, the mechanism 312 that pumps can include motor And cam, the motor cam is fixed on the output shaft of motor, and cam is directly mutually compressed with swinging base 21 or through can be The push rod moved up and down in guide groove is mutually compressed or is hinged.The motor of mechanism 312 of pumping is placed in pedestal 32.

The mechanism 312 that pumps can be one or more.Under normal circumstances, the machine that pumps Structure 312 can meet swing demand.Design is waved based on such, can be by adjusting the upper and lower past of mechanism that pump Multiple motion frequency, or pump the distance between mechanism and hinged seat is adjusted, waving for shaking table can be adjusted easily Amplitude and frequency.

During work, under Motor drive, the mechanism 312 that pumps promotes swinging base 21 to wave fortune around hinged seat Dynamic, and then drive the culture bag bracket 1 being fixed on support arm 211 to drive cell culture bags 4 to rock.Bracket passes through heating plate pair Cell culture is heated, and can be accessed respectively by being arranged at the accessory fixed hole position 111 of bracket base in cell culture bags Monitoring probe is planted, cell culture state is detected.In incubation, can also stop rocking, insert in the object lens insertion breach 112 of bracket Enter the direct observation of cell situation of object lens.

Embodiment 2 is used for the dark field microscope that cell culture is observed online

A kind of dark field microscope that is observed for cell culture online, microscope are also disclosed referring to Fig. 6, the present embodiment Including but not limited to：Illumination component, Moveable support 9, support base 5, object lens 61, photo-sensitive cell 10, caliber unit 6, A/ D conversion unit, control module.Illumination component is arranged at the top of Moveable support 9, Moveable support 9 is arranged on support bottom On seat 5, caliber unit 6 is arranged on 9 bottom of Moveable support, and object lens 101 are arranged on caliber unit 6 and corresponding in illumination unit The lower section of part, photo-sensitive cell 10 are arranged on the light signal in caliber unit 6 for gathering on object lens 61, and A/D converting units connect Photo-sensitive cell 10 and support base 5 is connect, control module connects A/D converting units.

Photo-sensitive cell adopts ccd sensor in the present embodiment, the image letter that ccd sensor is obtained by A/D converting units Number pass to control module.Control module also include setting in a computer check unit, for checking object lens under current state The content for being observed, and storage element, need the content for observing for preserving for storing.

In the present embodiment, illumination component is arranged on the top of Moveable support 9, and illumination component is including but not limited to justified The lamp socket 8 of shape and it is arranged on the lamp socket 8 of circle and surrounds the LED of 5 concentric arrays, the distance of lamp socket to object lens is 18 centimetres, general could be arranged to 10-30 centimetre.Include ten to 20 LEDs on each concentric circular, be whole dress Offer light source is provided.Lamp on each concentric circular on circumference can be opened simultaneously or be closed.

As shown in fig. 7, for the illumination for meeting dark field, light and the angle of 61 trunnion axis of object lens that illumination component sends It is less than angle A, angle of the angle A for the light and 61 trunnion axis of object lens in the bosom of illumination component.

In the present embodiment, support base 5 includes motor 501, slide mechanism and base 503, and slide mechanism includes Slide block 502 and slide rail, slide block 502 are connected on base 503 through slide rail, and 501 link control module of motor is simultaneously located at bottom One end of seat 503, slide block 502 connect motor 501, and Moveable support 2 is arranged on slide block 502.This area common Technical staff knows that motor 501 can be can achieve using motor 501 common in the market, motor 501 With the movement of movable slider 502 and then the relative position of entirely microscopical Moveable support 9 can be changed.

In the present embodiment, caliber unit 6 is slidably connected at 9 on Moveable support, and caliber unit 6 connects packaged type Chute is provided with support 9, caliber unit 6 can be slided up and down along chute, on Moveable support, 9 relative both sides are arranged Through hole, is respectively provided with screw on two through holes, when needing to adjust rising caliber unit 6, can unclamp screw, mobile caliber list Unit 6, when being adjusted to appropriate location, tightens screw.Certainly the simply trickle regulation of the regulation of this rising and reduction, maximal regulated Scope is 5 centimetres.Can certainly be used for playing fixed effect using the device of known similar buckle on other markets.When So, control module 7 can also be utilized as needed to control the switch of LED, to meet the needs of observation.

In another embodiment, Moveable support 9 has hollow structure, the illumination component control unit of control unit 7 Arrange in hollow structure.

Those skilled in the art understands, and module controlled as described above is obtained with regard to process that illumination component is controlled, CCD The process of the movement of the process and control support base 5 of the picture signal conversion for obtaining can utilize calculating of the prior art Machine, integrated circuit modules, PLD, other hardware or existing software module are realizing.

Using this device when sample is observed, with big advantage.During observation sample, can using the movement of control module 7 Removable support 9 is placed in whole culture vessel between illumination component and object lens 61, and object lens 61 pass through cell culture table object lens Insertion breach 112 carry out observation of cell, using control module move Moveable support 9, make it away from culture vessel, when need again The position of movable supporting frame 9 is then moved using control module during secondary observation.

3 cell culture bags of embodiment

As shown in FIG. 8 and 9, cell culture bags 4, including：Closed bag 41 and the fixed component 42 located at bag both sides, 41 top surface of closed bag is provided with outside liquid adding mouth 411, outside liquid injection port 412, gas feed 413 and gas Outlet 414,41 bottom surface of closed bag is provided with liquids recovery mouth 415 and at least one sensor connector 417, the outside It is all provided with liquid adding mouth 411, outside liquid injection port 412, gas feed 413, gas outlet 414 and liquids recovery mouth 415 There is aseptic snap joint 418.

4 top surface of cell culture bags is provided with outside liquid adding mouth, outside liquid injection port, gas feed and gas outlet Even if in order in the case of meeting and being not to move out culture apparatuses during cell culture, can also meet interpolation cell culture fluid, Outside liquid (such as penicillin solution, Streptomycin Solution, inoculation liquid etc.), and edema caused by disorder of QI (the such as offer needed for cell culture is provided Carbon dioxide) needs.Cell culture bags bottom surface is provided with liquids recovery mouth can be in the case where culture apparatuses be not to move out, also can be complete Change culture fluid in portion or part.Cell culture bags bottom surface is provided with sensor connector, can be used to connect sensor to monitor cell training Index of correlation in nutrient solution bag.

Snap joint refers to that the instrument of not needing can be achieved with the joint that pipeline is switched on or switched off.Aseptic snap joint used is For conventional aseptic snap joint, commercially available acquisition.Aseptic snap joint is solderable or is welded on bag.

Described closed bag can be made up of flexible material.The flexible material refers to deformable material after stress Matter.Preferably, material used by the closed bag is composited by PE and EVOH.

In a preferred embodiment, the material used by the closed bag is multi-layer compound structure.Preferably, the envelope Enclosed bag innermost layer is polyethylene (PE) layer.Innermost layer is that layer of abutment culture fluid, and contact culture fluid is stable PE materials, low leachable, without animality originate.

In embodiment is more highly preferred to, material used by the closed bag is five layers of composite construction, from the inside to the outside according to Secondary for polyethylene (PE) layer, adhesive layer, ethylene propylene enol (EVOH) layer, adhesive layer and polyethylene (PE).Such five layers of composite junction The material of structure be prior art, commercially available acquisition.

The bag webs thickness for constituting sealed bag can be conventional, employ 0.325mm in a preferred embodiment.This practicality New cell culture bags can be carried out disinfection sterilizing with gamma-rays, irradiation dose >=25Gy ,≤50Gy.The method of available heating is entered Row sealing.

In this utility model preferred embodiment, the closed bag by upper bag sheet 4161, lower bag sheet 4162, left Side pocket piece 4163 and right side bag webs 4164 surround a stereoscopic bag chamber jointly.More preferred, the left side bag webs 4163 and described The bottom of right side bag webs 4164 is curved.Base cambered design can cause the 3D shapes that cell culture bags are formed after inflation to exist Shearing force produced by the shake of cultivation platform is less, and the shearing force damage caused by cell is also down to minimum.In such as Figure 10 In shown embodiment, 4164 equal ovalize of the left side bag webs 4163 and the right side bag webs, 4161 He of the upper bag sheet Lower bag sheet 4162 is rectangular.

In preferred embodiment as shown in Figure 9, transparent observing area 419 in the middle part of the bottom of the bag 41, is provided with.Transparent sight Cha Qu, can be direct from bottom by setting cellular morphology finder on cultivation platform located at the bottom of cell culture bags Cell situation in observation of cell culture bag.

Described sensor connector can be provided with one or more.Preferably, the preferred embodiment as shown in Fig. 9 and 12 In, two sensor connector are provided with, one of them is the sensor connector 4171 for accessing pH sensor probe, another It is the sensor connector 4172 for accessing dissolved oxygen (DO) sensor probe.

It can also be contactless sensor connector that sensor connector can be the sensor connector of contact.Preferred real Apply in example, sensor connector is contactless sensor connector, i.e. sensor probe after the sensor connector is inserted, and visits Head with bag in culture fluid not in contact with.

In preferred embodiment as shown in figure 12, the contactless sensor connector includes basal disc as shown in figure 13 41701 and sensor probe containing cavity 41702, transparent cut-off 1703 is provided with the middle part of the basal disc, the sensor probe holds Put chamber 1702 to be connected with the basal disc 41701 and the side positioned at the transparent cut-off 41703, the opposite side of the transparent cut-off (pointing to referring to shown by arrow) is provided with conversion film.To being used for the sensor connector for accessing pH sensor probe, the conversion film Color-changing membrane is caused for pH.PH causes color-changing membrane be obtained by masking after pH indicators and filmogen mixture.The pH causes color-changing membrane category PH sensing membranes used in prior art, such as pH electrode etc..To being used for the sensor connector for accessing dissolved oxygen sensor probe, institute State film is changed into dissolved oxygen cause color-changing membrane.Dissolved oxygen causes color-changing membrane be obtained by masking after dissolved oxygen indicator and filmogen mixture ?.The dissolved oxygen causes color-changing membrane to belong to prior art.

Through the sensor connector, in cell culture bags, the pH of culture fluid and dissolved oxygen situation are both transferred to color letter Number, by accessing RGB sensors in sensor connector, you can read color signal, so as to according to conversion film color and pH Value and the relation of dissolved oxygen, obtain the pH value and oxygen dissolving value of cell culture fluid.The result of sensor connector detection is can transmit to control Molding block, for recording and checking.

The RGB color sensor that noncontacting proximity sensor probe is used cooperatively not only is operated more square without the need for sterilization And can more preferable Pollution protection just,.

The sensor connector of the contact can be normal sterile snap joint, can directly grafting or threaded connection sterilize PH sensors or the probe of dissolved oxygen sensor.Aforementioned noncontacting proximity sensor joint can also be made after removing transparent cut-off and conversion film Sensor connector for contact.

Described fixed component 42 is used for coordinating the fixation cell culture bags in cultivation platform, can adopt existing Various can make the parts that Sheng liquid bag is fixed realizing the fixation of cell culture bags, can for example be single use can achieve solid Fixed part, such as with sticking part etc., but also need to play admittedly with the coalignment cooperation arranged on cultivation platform The part being set for, the part that is for example fixed by mechanical force, viscous force, magnetic force etc..In embodiment as shown in Figure 10, described solid Part is determined for hold-down bars, the hold-down bars can be removable with the fixed rod clamp set up on cultivation platform or hold-down bars interposer Formula connection is unloaded, such that it is able to reach the purpose of immobilized-cell culture bag.

Further, the two ends of the bag 41 are provided with seal area 44, and the hold-down bars is placed in seal area.

Cell culture bags of the present utility model can be fixed on cultivation platform by fixed component 42 when using, will be thin The outside liquid adding mouth 411 of born of the same parents' culture bag, outside liquid injection port 412, gas feed 413, gas outlet 414, liquid are returned 415 are closed up by aseptic snap joint, is connected with the device of offer outside liquid, gas or withdrawal liquid, gas by the road respectively Connect, the probe of grafting respective sensor in the sensor connector.During cultured cells, add as needed at any time outer Portion's liquid, changes culture fluid, there is provided appropriate cell culture atmosphere, and monitors the situation of cell culture fluid and the growth feelings of cell Condition.

Specifically, the outside liquid adding mouth 411 can be as shown in figure 11, through 418 rapid abutting joint of aseptic snap joint outside Portion's pipeline, the exterior line can include silica gel tube 431, female Luer 432 successively, and then externally connected liquid provides dress Put, threeway 433 can be accessed to connect multichannel outside liquid memory element on further silica gel tube.

The outside liquid injection port 412 can be as shown in figure 11, through 18 rapid abutting joint exterior line of aseptic snap joint, institute Stating exterior line can include silica gel tube 431, female Luer 432, needleless sampler 434 successively, and then connect outside liquid offer Device.

The gas feed 413 can be as shown in figure 11, through 418 rapid abutting joint exterior line of aseptic snap joint, described outer Portion's pipeline can include silica gel tube 431, air filter 435 successively, and then connect extraneous gas offer device.

The gas outlet 414 can be as shown in figure 11, through 418 rapid abutting joint exterior line of aseptic snap joint, described outer Portion's pipeline can include silica gel tube 431, air filter 435, silica gel tube 431, air cutoff valve 436 successively, so with extraneous or Gas collecting apparatus is communicated.

The liquids recovery mouth 415 can be as shown in figure 12, through 418 rapid abutting joint exterior line of aseptic snap joint, described Exterior line can include silica gel tube 431 and female Luer 432 successively, and then connect outside liquid collector.

Piece folder 437 can be set on the silica gel tube of pipeline to control to pass in and out sample, pipeline components can pass through the sealing of band 438 and connect Connect.

With regard to the cell culture bags in of the present utility model, following cell cultivation requirement can be met：

(1) meet and be changed without continuous condition of culture：Cell starting culture, amount are less, with Cell Culture Cells density Increase, need to uninterruptedly increase cell culture fluid；It is less that cell culture bags of the present utility model can meet initiator cell culture amount In the case of, also can be cultivated in cell culture bags；

(2) temperature conditionss of cell culture are met：No matter the initial amount of cell culture is how many, all can be with of the present utility model Cell culture bags are cultivated, and can carry out heat treatment to cell culture bags；

(3) conditions such as cell culture PH, dissolved oxygen (DO) are met：The design of 3D culture bags, can connect cell culture necessary Gas device, and by adjusting the revision that gaseous environment carries out the pH value and dissolved oxygen of culture fluid；

(4) non-contacting PH in cell cultivation process, dissolved oxygen detection are met：Special harmless by arranging in culture bag Avirulent photochemical reaction material, through the monitoring that cell culture bags carry out the PH of culture fluid, dissolved oxygen (DO)；

(5) meet the aseptic condition of cell culture：Total interface in culture bag, culture fluid import, the interface of recovery tube, gas Body is imported and exported, and all carries out aseptic rapid abutting joint mode, and gas inlet and outlet is fitted with pneumatic filter, it is ensured that cell culture is in Aseptic condition.

(6) in the case where cell culture system is rocked, the shearing force suffered by cell in culture bag is also less, favorably Growth in cell.

4 control platform of embodiment

In the present embodiment, control platform includes that control platform includes switch board 13, and is arranged on switch board 13 Automatic heating unit, preheating bag 1309 and peristaltic pump；Automatically heating unit includes preheating table, and the preheating bag 1309 is arranged on pre- On hot plate, peristaltic pump includes the first peristaltic pump 1301 and the second peristaltic pump 1302, and the entrance for preheating bag 1309 passes through the first pipeline 1308 connection external cellular culture fluid 1306, the outlet for preheating bag 1309 connect cell culture bags by second pipe 1307, the One peristaltic pump 1301 connects the first pipeline 1308, and the second peristaltic pump 1302 connects second pipe 1307, automatic heating unit, first 1302 link control module of peristaltic pump 1301 and the second peristaltic pump.

In the present embodiment, switch board 13 is exactly have well-regulated cube structure, can house various work inside which Unit and integrated circuit.Wherein, preheating table is arranged at the top of switch board 13, arranges preheating bag 1309 thereon.The work of preheating table Be bag 1309 to preheating in cell culture fluid heating.Cell culture fluid is suitable for this cell training using sold on market Model needed for supporting.The volume of preheating bag 1309 is traditionally arranged to be 500ml.Automatic heating unit link control module, so can To set a certain specific temperature, when the temperature of the cell culture fluid preheated in bag 1309 is less than this temperature, will be automatic Heated to preheating bag 1309 using preheating table, when reaching this temperature, heating stops.First peristaltic pump 1301 is by outside cell Culture fluid pumps into preheating bag 1309, and the cell culture fluid for reaching a certain specified temp pumps into cell training by the second peristaltic pump 1302 Foster bag.

Gas mixing units are additionally provided with the inside of switch board 13, the source of the gas outside gas mixing units connection, mainly Oxygen, air and carbon dioxide.Extraneous gas enter gas mixing units be mixed after through gas outlet 1305, flow through the 3rd Cell culture bags are entered after pipeline (not shown).Gas mixing units link control module 7, controls gas by control module 7 Component ratio of the uninterrupted of the supply of body, such as gas, the time of conveying and various gases etc..

In the present embodiment, pH value detection tabs 1303 and dissolved oxygen detection tabs 1304 are additionally provided with switch board, and all Sensor connector in difference link control module 7 and cell culture bags, by the pH value measured in cell culture bags and dissolved oxygen The signal transmission of amount is to control module 7.PH detection tabs 1303 and dissolved oxygen detection tabs 1304 all adopt on market commonly use Signal transmission joint, for example with SMA905 plugs.PH detection tabs 1303 and dissolved oxygen detection tabs 1304 and cell culture Connected using signal transduction part (not shown) between bag, such as common light connecting line on market.

5 control module of embodiment

Referring to Figure 14, for further optimizing design scheme, this utility model also includes that control module 7, control module 7 set Put on computers, including but not limited to illumination component control unit；Moveable support connects control unit, such as user The displacement that motor can be controlled and then control Moveable support 9 by being input into specific numerical value；Check unit, The person of being used primarily for checks every ginseng of the content and cell growth environment being observed that under current state by object lens Number；Storage element, is mainly used in storage user and thinks in the content that observes and the cell growth process for need preservation Ambient parameter, such as pH value and dissolved oxygen etc., in case check again；Gas control unit, is mainly used in controlling gas mixing list Unit, extraneous gas is input into after mixing cell culture bags when needing to take a breath；Culture fluid control unit, Main Function are to work as When cell culture fluid in cell culture bags is needed with changing, outside cell culture fluid is heated, and is input into cell culture Bag.

User can be controlled whole system and check and record each in various cell culture by control unit 7 Plant structure of parameter and cell etc..

Those skilled in the art understand, and above-mentioned control module is for the control of Moveable support as above Journey, the control process to illumination component, check the content and various parameters of cell culture bags process, control external air source with And cell culture fluid enter cell culture bags process, can utilize computer of the prior art, integrated circuit modules, can Programmed logic device, other hardware or existing software module are realizing.

Above-described embodiment only illustrative principle of the present utility model and its effect are new not for this practicality is limited Type.Any person skilled in the art all can be carried out to above-described embodiment without prejudice under spirit and the scope of the present utility model Modifications and changes.Therefore, such as those of ordinary skill in the art without departing from the essence disclosed in this utility model All equivalent modifications completed under god and technological thought or change, should be covered by claim of the present utility model.

Claims (25)

1. a kind of full-automatic cell continuous culture system, it is characterised in that the cell culture system at least includes：Control mould Block, control platform, dark field microscope, cell culture table, cell culture bags and the power supply observed for cell culture online Module；

The cell culture table includes cultivating bag bracket, rocking carrier and agitating device, and the culture bag bracket is provided with arcuate bottom Plate and cell culture bags fixed component, the rocking carrier include swinging base, and the swinging base is provided with support arm, the training Foster bag bracket is fixedly connected through support arm with the rocking carrier, and the agitating device includes that pedestal, pedestal are provided with and wave component, The swinging base is connected with component is waved, and the curved plate is provided with object lens insertion breach；

The cell culture bags, including closed bag, are provided with transparent observing area in the middle part of the bottom surface of the bag；The bag is put In cell culture table, the transparent observing area is mated with object lens gap position；

The dark field microscope that is observed for cell culture online, including object lens, the object lens just can be inserted into cell training Object lens insertion breach on foster shaking table；

The control platform is connected to the cell culture dark field microscope of observation and cell culture table online respectively；

The control module connects control platform；

The power module is powered for whole system.

2. full-automatic cell continuous culture system according to claim 1, it is characterised in that：Described exist for cell culture The dark field microscope of line observation includes illumination component, Moveable support, support base, photo-sensitive cell, caliber unit, A/D Converting unit, the illumination component are arranged on the top of Moveable support, and the Moveable support is arranged on support base On, the caliber unit is connected to Moveable support bottom, and the object lens are arranged on caliber unit and corresponding in illumination unit The lower section of part, the photo-sensitive cell are arranged on the light signal in caliber unit for gathering on object lens, the A/D converting units Connection photo-sensitive cell, the control module connect A/D converting units.

3. full-automatic cell continuous culture system according to claim 2, it is characterised in that：The illumination component includes justifying The lamp socket of shape, and be arranged on lamp socket the illuminating lamp for surrounding several concentric arrays, the center of circle of the concentric circular and lamp socket Home position consistent.

4. full-automatic cell continuous culture system according to claim 3, it is characterised in that：The lamp socket to object lens away from From for 10-30 centimetre.

5. full-automatic cell continuous culture system according to claim 4, it is characterised in that：The light source of the illuminating lamp with Object lens form dark field, meet the needs of dark field illumination.

6. full-automatic cell continuous culture system according to claim 3, it is characterised in that：The illuminating lamp is LED.

7. full-automatic cell continuous culture system according to claim 2, it is characterised in that：The support base includes walking Stepper motor, slide mechanism and base, the slide mechanism include that slide block and slide rail, the control module connect and control stepping electricity Machine, the motor connection sliding block, the slide block be slidably connected at through slide rail on base and horizontally moveable, described removable Formula support is arranged on slide block.

8. full-automatic cell continuous culture system according to claim 1, it is characterised in that：Arrange on the curved plate One or more accessory fixed hole positions, leave under the accessory fixed hole position for installing or insert the space of outside appurtenances.

9. full-automatic cell continuous culture system according to claim 1, it is characterised in that：The curved plate is provided with Heating plate.

10. full-automatic cell continuous culture system according to claim 1, it is characterised in that：The side of the curved plate Edge is provided with baffle plate.

11. full-automatic cell continuous culture systems according to claim 1, it is characterised in that：The cell culture bags are solid Part is determined located at the both sides of curved plate, including the platform extended from two lateral both sides of curved plate, the platform is provided with solid Determine rod clamp and/or hold-down bars interposer.

12. full-automatic cell continuous culture systems according to claim 1, it is characterised in that：The swinging base is flat Plate, two ends are provided with support arm, and culture bag bracket is not contacted with swinging base, between leave space to install or match somebody with somebody outside inserting Part.

13. full-automatic cell continuous culture systems according to claim 1, it is characterised in that：The component that waves includes Hinged seat and at least one pumps mechanism, and on pedestal, the swinging base is cut with scissors the hinged seat with hinged seat Connect, the mechanism that pumps is connected with swinging base or is mutually compressed.

14. full-automatic cell continuous culture systems according to claim 1, it is characterised in that the cell culture bags bag Include：The fixed component of bag both sides, the closed upper face of bag body are provided with outside liquid adding mouth, outside liquid injection port, gas Body import and gas outlet, the closed bag bottom surface are provided with liquids recovery mouth and at least one sensor connector, described outer It is equipped with portion's liquid adding mouth, outside liquid injection port, gas feed, gas outlet and liquids recovery mouth and aseptic quickly connects Head.

15. full-automatic cell continuous culture systems according to claim 14, it is characterised in that：The closed bag by Upper bag sheet, lower bag sheet, left side bag webs and right side bag webs surround a stereoscopic bag chamber, the left side bag webs and the right side bag webs jointly Bottom curved, the left side bag webs and the equal ovalize of the right side bag webs, the upper bag sheet and lower bag sheet rectangular.

16. full-automatic cell continuous culture systems according to claim 14, it is characterised in that：The closed bag Innermost layer is polyethylene layer.

17. full-automatic cell continuous culture systems according to claim 14, it is characterised in that：The closed bag institute Material is five layers of composite construction, is followed successively by polyethylene layer, adhesive layer, ethylene, propylene alcohol layer, adhesive layer from the inside to the outside and gathers Pvdf layer.

18. full-automatic cell continuous culture systems according to claim 14, it is characterised in that：Described sensor connector It is provided with one or more.

19. full-automatic cell continuous culture systems according to claim 18, it is characterised in that：Described sensor connector For noncontacting proximity sensor joint or touch sensor joint.

20. full-automatic cell continuous culture systems according to claim 19, it is characterised in that：The contactless biography Sensor joint includes that basal disc and sensor probe containing cavity, the basal disc are provided with transparent cut-off, and the sensor probe is housed Chamber is connected with the basal disc and the side positioned at the transparent cut-off, and the opposite side of the transparent cut-off is provided with conversion film.

21. full-automatic cell continuous culture systems according to claim 20, it is characterised in that：The film of changing is caused into pH Color-changing membrane or dissolved oxygen cause color-changing membrane.

22. according to the full-automatic cell continuous culture system described in claim 1, it is characterised in that：The control platform bag Include switch board, and automatic heating unit, preheating bag and the peristaltic pump being arranged on switch board；The automatic heating unit includes Preheating table, the preheating bag are arranged on preheating table, and the peristaltic pump includes the first peristaltic pump and the second peristaltic pump, the cell By the cell culture fluid outside the first pipeline communication, the outlet of the preheating bag is connected by second pipe for the import of preheating bag Cell culture bags, first peristaltic pump connect the first pipeline, and second peristaltic pump connects second pipe, the automatic heating Unit, the first peristaltic pump and the second peristaltic pump link control module.

23. according to the full-automatic cell continuous culture system described in claim 22, it is characterised in that：Inside the switch board Gas mixing units are additionally provided with, the gas outlet of the gas mixing units is by the 3rd pipeline communication cell culture bags, the gas The air inlet connection external air source of body mixed cell, the gas mixing units link control module.

24. according to the full-automatic cell continuous culture system described in claim 22, it is characterised in that：On the switch board also It is provided with pH value detection tabs and dissolved oxygen detection tabs, the pH value detection probe and dissolved oxygen detection probe link control module And cell culture bags.

25. according to the full-automatic cell continuous culture system described in claim 1, it is characterised in that the control module bag Include：

Moveable support control unit, for controlling the movement of Moveable support；

Illumination component control unit, for controlling the switch of illumination component；

Unit is checked, for checking object lens are observed under current state content, and cell growth environment under current state Parameters；

Storage element, for storing the parameters of the content for the observing and record cell growth environment for needing to preserve；

Gas control unit, for controlling the conveying of the gas in cell culture bags；

Cell culture fluid control unit, for controlling temperature and the supply of cell culture fluid.