CN108504568B - Stem cell automatic amplification culture equipment based on modularization - Google Patents

Stem cell automatic amplification culture equipment based on modularization Download PDF

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Publication number
CN108504568B
CN108504568B CN201810168377.6A CN201810168377A CN108504568B CN 108504568 B CN108504568 B CN 108504568B CN 201810168377 A CN201810168377 A CN 201810168377A CN 108504568 B CN108504568 B CN 108504568B
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cover
culture
fixedly arranged
stage
seat
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CN108504568A (en
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谭映军
李莹辉
王春艳
聂捷琳
施镠佳
顾寅
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SPACENTER SPACE SCIENCE AND TECHNOLOGY INSTITUTE
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Space Institute Of Southern China (shenzhen)
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    • C12M29/00—Means for introduction, extraction or recirculation of materials, e.g. pumps
    • C12M29/18—External loop; Means for reintroduction of fermented biomass or liquid percolate
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    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/48—Automatic or computerized control

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Abstract

The invention discloses a stem cell automatic amplification culture device based on modularization, wherein a sample frame capable of rotating in a core component of the device comprises a primary culture module and a secondary culture module which are clamped in a module bracket, cell culture bags in the culture modules are connected in series through a drainage needle, a micropump, a needle seat and a pipeline, liquid in two adjacent cell culture bags after being connected in series can be uniformly distributed after being circularly mixed under the action of the micropump, and a control execution system is provided with a camera, a temperature sensor, a CO (carbon monoxide) and a control circuit board which are connected with the control circuit board2The device comprises a sensor, a fan, a radiator, a heater and an electromagnetic valve, wherein the electromagnetic valve is communicated with a gas pipe; the device adopts the cell culture module that can expand automatically to carry out the automatic amplification culture of individualized stem cell, has integrateed culture environment temperature and gaseous composition automatic control system, can realize stem cell culture alone, and the control to the cell growth proliferation condition can be realized to integrated cell proliferation density detection sensor.

Description

Stem cell automatic amplification culture equipment based on modularization
Technical Field
The invention relates to stem cell amplification equipment, in particular to modularized stem cell amplification equipment which is based on personalized stem cell amplification culture, can automatically control the cell amplification culture environment and can monitor key indexes in real time on line.
Background
In the current research fields of clinical medicine, life science, cell biology and the like, the mass amplification and cultivation of stem cells is very important work, and particularly in the processes of treatment, beauty treatment, repair and the like based on stem cells, the requirements on individuation and customization of the stem cell amplification and cultivation are higher.
The existing stem cell culture equipment on the market at present does not have the capacity of self-regulation and control of a cell culture environment, and the normal work of the equipment also depends on auxiliary equipment such as a cell culture box, so that the existing equipment is influenced in the aspects of operability, application field and the like. The existing stem cell culture equipment does not have an automatic amplification function at present, and is restricted in standardization, scale and automation of stem cell amplification culture application; and because the existing stem cell culture equipment is not provided with a perfect on-line parameter monitoring system, the amplification and culture processes of the stem cells cannot be monitored in real time.
Disclosure of Invention
In order to solve the technical problems, the invention provides a stem cell automatic amplification culture device based on modularization, which can perform culture of stem cell automatic amplification culture, a cell culture environment temperature and gas composition automatic control system integrated with the device enables the stem cell culture to be realized without depending on other peripheral auxiliary devices, and a cell proliferation density detection sensor integrated with the device can realize monitoring of cell growth and proliferation conditions.
The technical scheme of the invention is as follows:
the invention discloses a stem cell automatic amplification culture device based on modularization, which comprises the following components:
the heat preservation cover can be opened on the base by more than 90 degrees and can be locked on the base, a touch screen is arranged on the heat preservation cover, a control circuit board is arranged in the base, and the touch screen is connected with the control circuit board;
the core component comprises a heat-insulation cover with an opening at the upper end and positioned in the base, a bottom plate is positioned in the heat-insulation cover, a sample supporting seat is fixedly arranged at the rear end of the bottom plate, a transmission seat is fixedly arranged at the front end of the bottom plate, a plurality of sample frames which are arranged in parallel are arranged in the heat-insulation cover, the sample frames are erected between the sample supporting seat and the transmission seat through sample frame rotating shafts, and the sample frames can rotate around the central shafts of the sample frames under the driving of a driving structure; the sample rack consists of a plurality of primary culture modules, a secondary culture module and a module bracket, wherein the primary culture module comprises a primary cell culture bag positioned in a primary accommodating cabin, a micropump and a needle seat; the first-stage cell culture bag and the second-stage cell culture bag are both provided with an impedance sensor; the first-stage culture module and the second-stage culture module are connected in series and then clamped in the module bracket;
the control execution system comprises a T-shaped partition plate which is fixedly arranged on the bottom plate and positioned below the sample rack, a camera is fixedly arranged at one end of the upper surface of a horizontal plate of the T-shaped partition plate, a small mounting seat is fixedly arranged at the middle position of one side of a vertical plate of the T-shaped partition plate, large mounting seats are fixedly arranged on two sides of one end of the vertical plate, and a temperature sensor and a CO (carbon monoxide) sensor are fixedly arranged on the small mounting seats2The sensor comprises a sensor and a wire holder, wherein a fan is fixedly arranged on the upper surface of the large mounting seat, a radiator is fixedly arranged on the bottom surface of the large mounting seat, and the radiatorA heater is fixedly arranged on the lower surface of the heater; an electromagnetic valve and a gas pipe are fixedly arranged on the other side of the vertical plate of the T-shaped partition plate, one end of the gas pipe is connected with an inlet of the electromagnetic valve, and the other end of the gas pipe is communicated with the gas cylinder after penetrating through the core component and the base; the camera, the temperature sensor and the CO2The sensor, the fan, the radiator, the heater and the electromagnetic valve are all connected with the control circuit board through the wire holders by leads.
The further technical scheme is as follows:
the first-stage cell culture bag and the second-stage cell culture bag comprise bag bodies, two connectors distributed at intervals are arranged at the upper ends of the bag bodies, two connectors distributed at intervals are arranged at the lower ends of the bag bodies, and the impedance sensors are positioned at the upper ends of the bag bodies.
The micro pump is fixedly arranged between the front end clapboard and the middle clapboard of the first-level lower cover, the needle seat is positioned and arranged on the front end clapboard of the first-level lower cover, the drainage needle is inserted on two connectors at the upper end of the first-level cell culture bag and is connected with the needle seat through the micro pump and a pipeline, and the needle seat is connected with a connector at the lower end of the previous first-level or second-level cell culture bag adjacent to the first-level cell culture bag.
The drainage needle head is connected with the needle head seat through a micropump and a pipeline in a mode that: one drainage needle is directly communicated with the needle seat by a pipeline, and the pipeline connected with the other drainage needle is communicated with the needle seat after passing through the micropump; or the pipelines connected with the two drainage needles are communicated with the two needle head seats respectively after passing through the same micropump.
The two ends of the first-stage upper cover and the two ends of the first-stage lower cover are positioned through positioning pins arranged on the first-stage lower cover, and the first-stage upper cover and the first-stage lower cover can be buckled to form a first-stage accommodating cabin; the second grade upper cover of module is cultivateed to the second grade and the one end of second grade lower cover is articulated, and the both ends of this second grade upper and lower cover are all fixed a position and make the second grade upper and lower cover can the lock form second grade holding cabin through the locating pin that sets up on the second grade lower cover.
The heat-insulation cover comprises an inner cover and an outer cover which are fixedly connected together in a laminated manner, wherein the surface of the inner cover is coated with a heat-insulation material in a sheet structure, the front edge of the inner cover is provided with a fastening lock in a positioning manner, and the hinge is arranged at the rear edge of the inner cover in a positioning manner; the touch screen is characterized in that the outer cover is provided with a mounting through hole and a mounting bottom plate, the touch screen is fixedly arranged on the mounting bottom plate, the mounting bottom plate is fixedly arranged at the mounting through hole, the touch screen is exposed out of the mounting through hole, and the touch screen is coated with a protective film.
The base includes a rear side open-ended protecting crust, and this protecting crust internal fixation cover is equipped with a core component mount pad, control circuit board sets firmly on the posterior lateral plate of core component mount pad, is equipped with a maintenance lid, and this maintenance lid detachable location is installed and is formed integratively and cover in the trailing flank of protecting crust and protecting crust control circuit board.
The driving structure in the core component comprises a motor arranged on the outer side of the transmission seat, and an output shaft of the motor extends into the inner side of the transmission seat and is positioned below the horizontal plate of the T-shaped partition plate; the sample rack is provided with a first synchronizing wheel and a second synchronizing wheel, the first synchronizing wheel is locked and positioned on an output shaft of the motor, the second synchronizing wheel is locked and positioned on a rotating shaft of the sample rack on the same side with the output shaft of the motor, and the first synchronizing wheel and the second synchronizing wheel are both positioned on the inner side of the transmission seat; the sample rack is provided with a first synchronous belt and a second synchronous belt, two ends of the first synchronous belt are respectively arranged on the first synchronous wheel and the second synchronous wheel, two ends of the second synchronous belt are respectively arranged on different second synchronous wheels, and the rotation of the output shaft of the motor drives the rotation shaft of the sample rack to rotate through the transmission of the first synchronous wheel, the second synchronous wheel, the first synchronous belt and the second synchronous belt.
The module support comprises an upper assembly and a lower assembly, one end of the upper assembly is hinged to one end of the lower assembly, and the other end of the upper assembly and the other end of the lower assembly can be locked and unlocked through a buckled locking piece.
The upper assembly comprises a T-shaped connecting block and a straight connecting block, two first supporting rods which are arranged at intervals are fixedly arranged between the T-shaped connecting block and the straight connecting block, a locking ring is fixedly arranged at the middle position of the inner side of the T-shaped connecting block, the lower assembly comprises a semicircular connecting block and an п -shaped connecting block, two second supporting rods which are arranged at intervals are fixedly arranged between the semicircular connecting block and the п -shaped connecting block, a locking buckle is fixedly arranged at the middle position of the inner side of the semicircular connecting block, the locking ring and the locking buckle can be locked and unlocked, the culture module is clamped between the first supporting rods and the second supporting rods, the rotating shaft of the sample frame comprises a long shaft and a short shaft, the long shaft is fixedly arranged at the middle position of the outer side of the semicircular connecting block, and the short shaft is fixedly arranged at the middle position of the outer side of the п -shaped connecting block.
The hinged connection structure of the upper and lower components is characterized in that a hinge is arranged, the left side of the hinge is fixedly arranged on the outer side of the straight-line-shaped connecting block, and the right side of the hinge is fixedly arranged on the outer side of the п -shaped connecting block.
The core component also comprises side plates fixedly arranged at the left side and the right side of the bottom plate and a transmission seat cover plate fixedly arranged at the outer side of the transmission seat; the sample rack is provided with a base ring and an outer plate ring, wherein openings capable of taking and placing sample racks are formed in the middle of the base ring and the middle of the outer plate ring, the base ring is fixedly arranged on the upper edges of a sample supporting seat, a transmission seat and a side plate, and the outer plate ring is fixedly arranged on the upper surface of the base ring; a sealing ring groove is formed in the outer plate ring along the opening of the outer plate ring, and a sealing ring is fixedly arranged in the sealing ring groove; and a maintenance handle is fixedly arranged at each of two ends of the base ring.
The beneficial technical effects of the invention are as follows: the culture equipment has the following technical characteristics:
1. the cell culture module with automatic amplification capability is adopted for stem cell culture, so that customized automatic amplification culture of the personalized stem cells can be carried out;
2. the automatic control system for the environmental temperature and the gas components of the cell culture is integrated, so that the stem cell culture can be realized without depending on other peripheral auxiliary equipment;
3. the cell proliferation density detection sensor is integrated, and the monitoring of the cell growth and proliferation conditions is realized.
Drawings
FIG. 1 is a schematic view of the overall structure of the present invention;
FIG. 2 is a schematic view of an exploded structure of the insulating cover of the present invention;
FIG. 3 is a schematic view of an exploded structure of the base of the present invention;
FIG. 4 is a schematic illustration of an exploded view of the core component of the present invention;
FIG. 5 is a schematic diagram of a control execution system according to the present invention;
FIG. 6 is a second schematic diagram of the control execution system according to the present invention;
FIG. 7 is a schematic view of the structure of a sample holder according to the present invention;
FIG. 8 is a schematic structural view of a module support of the present invention;
FIG. 9 is one of the schematic structural views of the primary culturing module of the present invention;
FIG. 10 is a second schematic structural view of the primary culturing module of the present invention;
FIG. 11 is a schematic diagram of the structure of a secondary culturing module of the present invention;
FIG. 12 is a schematic diagram showing the structure of the first and second culturing modules according to the present invention after they are connected in series;
FIG. 13 is a schematic view of the construction of the primary and secondary cell culture bags of the present invention;
wherein:
1-a housing;
11-a heat preservation cover; 111-inner cover; 112-outer cover; 113-insulation material; 114-snap lock; 115-mounting through holes; 116-a mounting base plate; 117-touch screen; 118-a protective film;
12-a base; 121-protective shell; 122-core component mount; 123-control circuit board; 124-maintenance cover;
13-a hinge;
2-a core component;
21-a heat preservation cover; 22-a base plate; 23-a sample holder; 24-a transmission seat;
25-a sample rack;
251-a module support;
251 a-upper assembly;
251a1-T connection block; 251a 2-straight connecting block; 251a3 — first support bar;
251 b-lower assembly;
251b 1-semicircular connecting block, 251b2- п type connecting block, 251b 3-second supporting rod;
251 c-a locking member;
251c 1-locking ring; 251c 2-locking buckle;
251 d-loose leaf;
252-a primary culturing module;
2521-a primary containment tank; 2521 a-front end baffle; 2521 b-intermediate baffle; 2521 c-rear end baffle;
2522-primary cell culture bag; 2523-a micropump; 2524-needle holder; 2525-a drainage needle; 2526-line;
2527-primary upper cover; 2528-primary lower cover; 2529-locating pin;
253-secondary culture module; 2531-a secondary containment tank; 2532-secondary cell culture bags; 2533-secondary upper cover;
2534-secondary lower cover;
254-bag body;
255-a linker;
256-impedance sensors;
26-rotation axis of sample holder; 261-long axis; 262-minor axis;
27-a drive structure; 271-a motor; 272-a second synchronizing wheel; 273-first timing belt; 274-a second synchronous belt;
28-side plate; 29-transmission seat cover plate; 210-a foundation ring; 211-outer plate ring; 212-sealing ring; 213-maintenance handle.
3-control the execution system;
a 31-T shaped spacer; 32-a camera; 33-small mounting seat; 331-a temperature actuator; 332-CO2A sensor; 333-to
A wire holder; 34-large mounting base; 341-a fan; 342-a heat sink; 343-a heater; 35-a solenoid valve; 36-gas delivery pipe.
Detailed Description
In order to make the technical means of the present invention clearer and to make the technical means of the present invention capable of being implemented according to the content of the specification, the following detailed description of the embodiments of the present invention is made with reference to the accompanying drawings and examples, which are provided for illustrating the present invention and are not intended to limit the scope of the present invention.
The invention relates to a modular stem cell automatic amplification culture device, which consists of a shell 1, a core component 2 and a control execution system 3.
The shell comprises a heat preservation cover 11 and a base 12 which are connected through a hinge 13, the heat preservation cover can be opened on the base by more than 90 degrees and can be locked on the base, a touch screen 117 is arranged on the heat preservation cover, a control circuit board 123 is arranged in the base, and the touch screen is connected with the control circuit board;
the heat preservation cover 11 comprises a protective film 118, a touch screen 117, a mounting base plate 116, an outer cover 112, a fastening lock 114, an inner cover 111, a heat preservation material 113 and a hinge 13, wherein the outer cover and the inner cover are integrally processed and formed by using an ABS material through a CNC technology, the overall thickness is 5mm, a mounting through hole 115 is formed in the outer cover, the heat preservation material with a sheet structure is formed by foaming a PC material, the thickness is 10mm, the length and width dimension is 300mm × mm, the heat preservation material is adhered to the middle position of the surface of the inner cover through 3M double-sided glue, a LL-66S type stainless steel fastening lock is installed at the front edge of the inner cover through 2M 2.5 screws, a TI-340-5.00 type torque damping hinge is installed at the rear edge of the inner cover through the mounting hole in a positioning mode, a 10-inch 16: 9 touch screen is fixed on the mounting base plate made of the ABS material through 4M 3 screws, the mounting base plate with the thickness of 8mm is fixed to the middle position of the inner side of the outer cover through 6M 3 screws, the touch screen is exposed from the mounting through hole 115 in the middle of the outer cover, the protective film made of a transparent PS material.
The base 12 includes a core mount 122, a rear-side open protective case 121, a maintenance cover 124, and a control circuit board 123. The protective shell and the core component mounting seat are integrally machined and formed by CNC technology by adopting ABS materials, the integral thickness is 5mm, and the core component mounting seat is fixedly sleeved at the middle position of the protective shell through 10M 3 screws; the control circuit board is arranged on the position of the rear side plate of the core component mounting seat through 4M 3 screws; adopt ABS material by CNC technique whole machine-shaping, whole maintenance lid of 5mm of thickness is installed in the trailing flank of protective housing to fix with 2M 4 screws, form an organic whole with the protective housing and cover control circuit board.
The test specimen storage device comprises a core component 2, a seal ring 212, an outer plate ring 211, a base ring 210, a specimen support seat 23, a specimen rack 25, a side plate 28, a heat preservation cover 21, a bottom plate 22, a driving structure 27 (comprising a motor 271, a first synchronous wheel (not visible in the drawing), a second synchronous wheel 272, a first synchronous belt 273 and a second synchronous belt 274), a transmission seat cover plate 29, a maintenance handle 213 and a transmission seat 24, wherein the specimen racks in the embodiment are two, the upper end of the heat preservation cover 21 is open and is located in a core component mounting seat of a base 12, the bottom plate is machined and formed by 2A12 aluminum alloy, the thickness is 10mm, the size is 600mm × mm, the bottom plate is fixed in the heat preservation cover, a control execution system 3 is fixed in the middle of the bottom plate 22 by 4M 4 screws, the specimen support seat 23 is machined and formed by 2A12 aluminum alloy, the size is 300mm × mm 190mm ×.5mm, the rear end of the bottom plate is fixed by 3M 3, the rear end of the bottom plate is fixed by the corresponding to a rotating and fixed by the rotating shaft of a linear sealing module, the linear sealing ring 26 mm, the linear sealing module is a linear sealing module, the linear sealing module is a linear sealing module, the linear sealing module is a linear sealing module, the linear sealing module is a linear sealing module, the linear sealing module is a linear sealing module, the linear sealing module is a linear sealing module, the linear sealing module is a linear sealing module, the linear sealing module is a linear sealing module, the linear sealing module is a linear sealing module, the linear sealing module is set for linear sealing module.
The module support comprises an upper assembly 251a and a lower assembly 251b, one end of the upper assembly is hinged with one end of the lower assembly, the other end of the upper assembly and the other end of the lower assembly can be locked and unlocked through a buckled locking piece 251c, the upper assembly comprises a T-shaped connecting block 251a1 and a straight connecting block 251a2, the T-shaped connecting block and the straight connecting block are machined and formed by 2A12 aluminum alloy, the thickness of the T-shaped connecting block is 5mm, two first supporting rods 251a 3mm stainless steel pipes are arranged between the T-shaped connecting block and the straight connecting block at intervals, the two first supporting rods are fixed together through the T-shaped connecting block and the straight connecting block by 2M 3 screws, a locking ring 251c1 is fixed at the middle position of the inner side of the T-shaped connecting block at the middle position of the T-shaped connecting block, the 5mm stainless steel locking ring is fixed to the middle of the T-shaped connecting block by 2M 3 screws, the upper assembly 251b is fixed to the middle of the semi-circular connecting block 251b by 2M 3 screws, the middle connecting block 251a semicircular connecting block assembly is fixed to the middle connecting block 251b of the T-shaped connecting block at the middle position of the middle connecting block, the semi-semicircular connecting block 251b is fixed to the middle connecting block 251b of the semi-semicircular connecting block 251 c-semicircular connecting block, the semi-semicircular connecting block 251 b-semicircular connecting block 251 c-semicircular connecting block, the semi-semicircular connecting block 251 c-semicircular connecting block, the semi-semicircular connecting block is fixed to the semi-semicircular connecting block-3-semicircular connecting block, the semi-semicircular connecting block-semicircular.
A primary culture module 252 of the invention comprises a primary upper cover 2527, a primary lower cover 2528, a positioning pin 2529, a primary cell culture bag 2522, a micro pump 2523, two drainage needles 2525, two needle holders 2524 and three sections of pipelines 2526. the micro pump 2523 is connected with a control circuit board 123. the primary upper cover and the primary lower cover are molded by organic glass molding, the wall thickness is 5mm, the outer diameter is 70mm, the primary upper cover and one end of the primary lower cover are hinged, both ends of the primary upper cover and the primary lower cover are positioned by positioning pins arranged on the primary lower cover and can be buckled to form a primary accommodating chamber, the diameter of the positioning pins is 1mm, the primary upper cover and the primary lower cover can be freely opened and closed, a front end clapboard 2521a, a middle clapboard 2521b and a rear end clapboard 2521c are arranged in the primary accommodating chamber 2521, the section size is 17mm × 17mm, four M2 screws are arranged in the front end clapboard 1a primary lower cover of the primary lower cover and a middle clapboard 2521a secondary upper needle holder, the primary upper needle holder is connected with a front end of the primary culture bag, the primary upper needle holder, the drainage needle holder is connected with two drainage needle holder, the drainage needle holder is connected with the front end of the drainage needle holder, the drainage needle holder is connected with the drainage needle holder, the drainage needle holder is connected with the drainage needle holder, the.
The secondary culture module consists of a secondary upper cover 2533, a secondary lower cover 2534, a secondary cell culture bag 2532 and a positioning pin 2529. Second grade upper cover and second grade lower cover all adopt organic glass mould pressing machine-shaping, and the wall thickness is 5mm, and the appearance diameter is 70mm, and this second grade upper cover is articulated with one end between the second grade lower cover, and the both ends of second grade upper and lower cover are all fixed a position and make the second grade upper and lower cover can accurate lock formation can freely open and shut the second grade holding cabin through setting up in the locating pin 2529 that covers under the second grade, and the diameter of this locating pin is 1 mm. The second-stage cell culture bag 2522 is placed in a second-stage accommodating cabin of the second-stage lower cover, and four joints of the second-stage cell culture bag are correspondingly arranged at corresponding positions of the front and rear end clapboards of the second-stage lower cover.
The first and second cell culture bags 2522, 2532 include a bag 254 having four connectors 255 and an impedance sensor 256. Wherein two standard Ruhr connectors and an impedance sensor are bonded to the upper end of the bag body in a heat sealing mode, and the other two connectors are bonded to the lower end of the bag body in a heat sealing mode. The bag body is made of PET, is formed by blow molding, has a volume of 250ml and a thickness of 0.2 mm.
The control execution system 3 comprises a camera 32, a T-shaped partition plate 31, a small mounting seat 33, a large mounting seat 34, a temperature sensor 331 and CO2The temperature sensor, the CO sensor and the temperature sensor are characterized in that the temperature sensor, the CO sensor, the heater 343, the radiator 342, the electromagnetic valve 35 and the air delivery pipe 36 are formed, the T-shaped partition plate is fixed to the middle of the bottom plate 22 through 4M 4 screws and located below the sample rack, a horizontal plate and a vertical plate of the T-shaped partition plate are both machined and formed by ABS materials, the thickness of the horizontal plate is 2mm, the thickness of the vertical plate is 10mm, the USB micro-optical camera is fixed to one end of the horizontal plate of the T-shaped partition plate through 2M 3 screws, the vertical plate of the T-shaped partition plate is fixed to the middle of the back of the horizontal plate through 4M 4 screws, the small mounting seat is bent through a 1Cr13 stainless steel plate with the thickness of 0.5mm, the upper surface size is 40mm × mm, the temperature sensor and the CO sensor are2The sensor and the wire holder are respectively fixed on the upper surface by 4M 2, the large mounting base is bent by a 1Cr13 stainless steel plate with the thickness of 0.5mm, the size of the upper surface is 70mm × 50mm, the large mounting base is fixed at one end of a vertical plate by 4M 3 screws, a 6010 type 5V direct current fan is fixed on the upper surface by 4M 3 screws, a 24V film type heater is 15mm × 40mm, the size of the 24V film type heater is 3M double-faced adhesive tape is pasted on the surface of an aluminum alloy radiator, the heater and the radiator are fixed on the bottom surface of the large mounting base by 4M 3 screws, a 12V miniature electromagnetic valve is fixed on the side surface of the vertical partition plate, one end of a polytetrafluoroethylene gas pipe with the diameter of 3mm is connected with an inlet of the electromagnetic valve, the other end of the polytetrafluoroethylene gas pipe with the diameter of 3mm is connected2The sensor, fan, heat sink, heater and solenoid are all connected to the control circuit board 123 by leads using the wiring 333.
The operation mode is as follows: injecting mixed solution of cells and culture solution into the primary cell culture bag by using an injector, wherein the mixed solution is about 250ml, and the joint of the primary cell culture bag in the primary culture module is arranged at the corresponding position of the lower cover of the primary culture module. And respectively inserting 2 drainage needles into the joints on the middle partition plate, installing 2 needle bases at corresponding positions of the partition plate at the front end of the culture module, and connecting pipelines among the drainage needles, the needle bases and the micropumps. The upper cover and the lower cover of the first level are buckled by the positioning pin to form a whole. 250ml of cell culture solution is respectively filled into the other 3 cell culture bags and respectively filled into 2 primary culture modules and 1 secondary culture module according to requirements.
After 3 primary culture modules and one secondary culture module are sequentially butted according to the sequence of primary culture, primary culture and secondary culture, the needle seat is automatically inserted into the corresponding culture bag joint, so that 4 cell culture bags are connected in series. The assembled 4 culture modules were fixed into a module holder.
Turning on a power switch, displaying a parameter input interface on the touch screen by the equipment, and inputting experiment parameters according to experiment requirements; after a start key of an input interface is pressed, the equipment enters an automatic stem cell amplification and cultivation working mode, the temperature of an automatic control core component area of the equipment is 36.5 +/-0.5 ℃, and the concentration of CO2 is 5% +/-0.5%. During the stem cell amplification and cultivation process, the sensor monitors the cell concentration in real time, when the cell concentration in the first cultivation module meets the requirement, the equipment automatically controls the micro pump to work, the cells and the culture solution in the first cultivation module are injected into the cell cultivation bag of the second cultivation module, meanwhile, the culture solution in the cell cultivation bag of the second cultivation module is driven by pressure and flows back into the cultivation bag in the first cultivation module, the cell density in the cell cultivation bags in the first and second cultivation modules is half of the density in the original first cultivation module, and the purpose of automatic amplification is achieved.
Similarly, when the cell density in the cell culture bags in the first culture module and the second culture module meets the requirement, the equipment automatically controls the second micropump to work, so that the cells in the cell culture bags in the first culture module and the second culture module are evenly distributed into the cell culture bags in the first culture module, the second culture module and the third culture module, and when the cell density meets the requirement, the equipment evenly distributes the cells in the first culture module, the second culture module and the third culture module into the cell culture bags in the first culture module, the second culture module, the third culture module and the fourth culture module, so that the automatic expansion of the cells in one culture bag to the four culture bags is realized, and the volume is expanded to 1000ml from the original 250 ml.
When the cell density in the four culture bags meets the requirement, the equipment is automatically stopped and the end of the experiment is prompted.
The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention, it should be noted that, for those skilled in the art, many modifications and variations can be made without departing from the technical principle of the present invention, and these modifications and variations should also be regarded as the protection scope of the present invention.

Claims (10)

1. A stem cell automatic amplification culture equipment based on modularization is characterized in that: the device consists of the following components:
the heat preservation cover can be opened on the base by more than 90 degrees and can be locked on the base, a touch screen is arranged on the heat preservation cover, a control circuit board is arranged in the base, and the touch screen is connected with the control circuit board;
the core component comprises a heat-insulation cover with an opening at the upper end and positioned in the base, a bottom plate is positioned in the heat-insulation cover, a sample supporting seat is fixedly arranged at the rear end of the bottom plate, a transmission seat is fixedly arranged at the front end of the bottom plate, a plurality of sample frames which are arranged in parallel are arranged in the heat-insulation cover, the sample frames are erected between the sample supporting seat and the transmission seat through sample frame rotating shafts, and the sample frames can rotate around the central shafts of the sample frames under the driving of a driving structure; the sample rack consists of a plurality of primary culture modules, a secondary culture module and a module bracket, wherein the primary culture module comprises a primary cell culture bag positioned in a primary accommodating cabin, a micropump and a needle seat; the first-stage cell culture bag and the second-stage cell culture bag are both provided with an impedance sensor; the first-stage culture module and the second-stage culture module are connected in series and then clamped in the module bracket;
the control execution system comprises a T-shaped partition plate which is fixedly arranged on the bottom plate and positioned below the sample rack, a camera is fixedly arranged at one end of the upper surface of a horizontal plate of the T-shaped partition plate, a small mounting seat is fixedly arranged at the middle position of one side of a vertical plate of the T-shaped partition plate, large mounting seats are fixedly arranged on two sides of one end of the vertical plate, and a temperature sensor and a CO (carbon monoxide) sensor are fixedly arranged on the small mounting seats2The sensor comprises a sensor and a wire holder, wherein a fan is fixedly arranged on the upper surface of the large mounting seat, a radiator is fixedly arranged on the bottom surface of the large mounting seat, and a heater is fixedly arranged on the lower surface of the radiator; an electromagnetic valve and a gas pipe are fixedly arranged on the other side of the vertical plate of the T-shaped partition plate, one end of the gas pipe is connected with an inlet of the electromagnetic valve, and the other end of the gas pipe is communicated with the gas cylinder after penetrating through the core component and the base; the camera, the temperature sensor and the CO2The sensor, the fan, the radiator, the heater and the electromagnetic valve are all connected with the control circuit board through the wire holders by leads;
the two ends of the first-stage upper cover and the two ends of the first-stage lower cover are positioned through positioning pins arranged on the first-stage lower cover, and the first-stage upper cover and the first-stage lower cover can be buckled to form a first-stage accommodating cabin; the two-stage upper cover of the two-stage culture module is hinged with one end of the two-stage lower cover, and the two ends of the two-stage upper cover and the two ends of the two-stage lower cover are positioned through positioning pins arranged on the two-stage lower cover, so that the two-stage upper cover and the two-stage lower cover can be buckled to form a two-stage accommodating cabin;
the heat-insulation cover comprises an inner cover and an outer cover which are fixedly connected together in a laminated manner, wherein the surface of the inner cover is coated with a heat-insulation material in a sheet structure, the front edge of the inner cover is provided with a fastening lock in a positioning manner, and the hinge is arranged at the rear edge of the inner cover in a positioning manner; the touch screen is characterized in that the outer cover is provided with a mounting through hole and a mounting bottom plate, the touch screen is fixedly arranged on the mounting bottom plate, the mounting bottom plate is fixedly arranged at the mounting through hole, the touch screen is exposed out of the mounting through hole, and the touch screen is coated with a protective film.
2. The modular based stem cell auto-expansion culturing device of claim 1, wherein: the first-stage cell culture bag and the second-stage cell culture bag comprise bag bodies, two connectors distributed at intervals are arranged at the upper ends of the bag bodies, two connectors distributed at intervals are arranged at the lower ends of the bag bodies, and the impedance sensors are positioned at the upper ends of the bag bodies.
3. The modular stem cell-based automated expansion culture apparatus of claim 2, wherein: the micro pump is fixedly arranged between the front end clapboard and the middle clapboard of the first-level lower cover, the needle seat is positioned and arranged on the front end clapboard of the first-level lower cover, the drainage needle is inserted on two connectors at the upper end of the first-level cell culture bag and is connected with the needle seat through the micro pump and a pipeline, and the needle seat is connected with a connector at the lower end of the previous first-level or second-level cell culture bag adjacent to the first-level cell culture bag.
4. The modular stem cell-based automated expansion culture apparatus of claim 3, wherein: the drainage needle head is connected with the needle head seat through a micropump and a pipeline in a mode that: one drainage needle is directly communicated with the needle seat by a pipeline, and the pipeline connected with the other drainage needle is communicated with the needle seat after passing through the micropump; or the pipelines connected with the two drainage needles are communicated with the two needle head seats respectively after passing through the same micropump.
5. The modular based stem cell auto-expansion culturing device of claim 1, wherein: the base includes a rear side open-ended protecting crust, and this protecting crust internal fixation cover is equipped with a core component mount pad, control circuit board sets firmly on the posterior lateral plate of core component mount pad, is equipped with a maintenance lid, and this maintenance lid detachable location is installed and is formed integratively and cover in the trailing flank of protecting crust and protecting crust control circuit board.
6. The modular based stem cell auto-expansion culturing device of claim 1, wherein: the driving structure in the core component comprises a motor arranged on the outer side of the transmission seat, and an output shaft of the motor extends into the inner side of the transmission seat and is positioned below the horizontal plate of the T-shaped partition plate; the sample rack is provided with a first synchronizing wheel and a second synchronizing wheel, the first synchronizing wheel is locked and positioned on an output shaft of the motor, the second synchronizing wheel is locked and positioned on a rotating shaft of the sample rack on the same side with the output shaft of the motor, and the first synchronizing wheel and the second synchronizing wheel are both positioned on the inner side of the transmission seat; the sample rack is provided with a first synchronous belt and a second synchronous belt, two ends of the first synchronous belt are respectively arranged on the first synchronous wheel and the second synchronous wheel, two ends of the second synchronous belt are respectively arranged on different second synchronous wheels, and the rotation of the output shaft of the motor drives the rotation shaft of the sample rack to rotate through the transmission of the first synchronous wheel, the second synchronous wheel, the first synchronous belt and the second synchronous belt.
7. The modular based stem cell auto-expansion culturing device of claim 1, wherein: the module support comprises an upper assembly and a lower assembly, one end of the upper assembly is hinged to one end of the lower assembly, and the other end of the upper assembly and the other end of the lower assembly can be locked and unlocked through a buckled locking piece.
8. The modular stem cell automatic amplification culture device according to claim 7, wherein the upper assembly comprises a T-shaped connection block and a straight connection block, two first support rods are arranged between the T-shaped connection block and the straight connection block at intervals, a locking ring is arranged at the middle position of the inner side of the T-shaped connection block, the lower assembly comprises a semicircular connection block and an п -shaped connection block, two second support rods are arranged between the semicircular connection block and the п -shaped connection block at intervals, a locking buckle is arranged at the middle position of the inner side of the semicircular connection block corresponding to the locking ring, the locking ring and the locking buckle can be locked and unlocked, the culture module is clamped between the first and second support rods, the rotating shaft of the sample holder comprises a long shaft and a short shaft, the long shaft is arranged at the middle position of the outer side of the semicircular connection block, and the short shaft is arranged at the middle position of the outer side of the п -shaped connection block.
9. The apparatus of claim 7, wherein the upper and lower members are hingedly connected to each other by a hinge having a left side fixed to the outer side of the I-shaped connecting block and a right side fixed to the outer side of the п -shaped connecting block.
10. The modular based stem cell auto-expansion culturing device of claim 1, wherein: the core component also comprises side plates fixedly arranged at the left side and the right side of the bottom plate and a transmission seat cover plate fixedly arranged at the outer side of the transmission seat; the sample rack is provided with a base ring and an outer plate ring, wherein openings capable of taking and placing sample racks are formed in the middle of the base ring and the middle of the outer plate ring, the base ring is fixedly arranged on the upper edges of a sample supporting seat, a transmission seat and a side plate, and the outer plate ring is fixedly arranged on the upper surface of the base ring; a sealing ring groove is formed in the outer plate ring along the opening of the outer plate ring, and a sealing ring is fixedly arranged in the sealing ring groove; and a maintenance handle is fixedly arranged at each of two ends of the base ring.
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Address after: No.2 and No.5 factories and dormitories of Longkou industrial park, No.3 Industrial Road, Gaoqiao Industrial Zone, Pingdi street, Longgang District, Shenzhen City, Guangdong Province

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Address before: No.2 and No.5 factories and dormitories of Longkou industrial park, No.3 Industrial Road, Gaoqiao Industrial Zone, Pingdi street, Longgang District, Shenzhen City, Guangdong Province

Patentee before: SPACE INSTITUTE OF SOUTHERN CHINA (SHENZHEN)