CN111690537B - Stem cell preparation system - Google Patents
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- CN111690537B CN111690537B CN202010637359.5A CN202010637359A CN111690537B CN 111690537 B CN111690537 B CN 111690537B CN 202010637359 A CN202010637359 A CN 202010637359A CN 111690537 B CN111690537 B CN 111690537B
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Abstract
The invention relates to a stem cell preparation system which comprises a stem cell preparation device, a digestive juice tank, a physiological saline tank, a supernatant collecting tank, a waste liquid collecting tank, a culture medium recovery tank and a stem cell collector. The stem cell preparation device comprises a shell, and a digestion chamber, a centrifugal chamber and a culture chamber which are sequentially connected from top to bottom and are positioned in the shell. The digestive juice box, the physiological saline box, the supernatant collecting box, the waste liquid collecting box, the culture medium recycling box and the stem cell collector are connected with the stem cell preparation device through pipelines. The invention has high integration level and high automation degree, does not need artificial participation in the stem cell culture process, and furthest reduces the influence of external bacteria on stem cell culture.
Description
Technical Field
The invention belongs to the technical field of stem cell preparation, and particularly relates to a stem cell preparation system.
Background
Stem cells are the origin cells. Briefly, it is a primitive undifferentiated cell type with multipotent differentiation and self-replication capacity, which is the primitive cell that forms various tissue organs of mammals. Stem cells share morphological commonalities, usually circular or oval, small cell volume, relatively large nuclei, mostly euchromatin, with higher telomerase activity. Stem cells can be classified into embryonic stem cells and adult stem cells.
Along with the improvement of the living standard of people and the development of medical science and technology, the stem cell transplantation technology is increasingly applied to the fields of enhancing immunity and resisting aging besides the aspect of disease treatment. In the prior art, in the stem cell preparation process, a plurality of process links are relatively independent, and products obtained in the previous process link are conveyed into the downward process link in a mode of long-distance pipeline or manual conveying in series connection, so that the control of the sterile environment is very unfavorable.
Disclosure of Invention
The invention aims to solve the technical problems that: the invention has the advantages of high integration level and high automation degree, and the stem cell culture process does not need to be manually participated, so that the influence of external bacteria on the stem cell culture is reduced to the greatest extent.
The invention solves the problems existing in the prior art by adopting the technical scheme that:
a stem cell preparation system comprises a stem cell preparation device, a control system, a power supply system, a digestive juice tank, a physiological saline tank, a supernatant collecting tank, a waste liquid collecting tank, a culture medium recovery tank and a stem cell collector.
The stem cell preparation device comprises a shell, and a digestion chamber, a centrifugal chamber and a culture chamber which are sequentially connected from top to bottom and are positioned in the shell.
The digestion chamber upper end face is equipped with adipose tissue and adds the mouth, and the digestion liquid case adds mouth through digestion liquid and adds mouth through the connection, and digestion chamber inside is equipped with kickboard I, and kickboard I upper end fixedly connected with digestion chamber supernatant discharge pipe, and digestion chamber supernatant discharge pipe and kickboard I contact position open has the hole, and digestion chamber supernatant discharge pipe and supernatant collecting box link up and are connected. A feed opening is arranged in the middle of the lower end surface of the digestion chamber, a filter screen is arranged at the upper end of the feed opening, and a sealing disc is arranged at the lower end of the filter screen.
The inside kickboard II that is equipped with of centrifugal chamber, kickboard II up end and centrifugal chamber supernatant discharge pipe one end fixed connection, centrifugal chamber supernatant discharge pipe and supernatant collecting box link up and are connected, and the protruding a plurality of stirring boards that are equipped with in the inside periphery of centrifugal chamber, centrifugal chamber bottom surface opening is sealed through circular shape closure plate II, and the outside cover of centrifugal chamber is equipped with belt pulley II, and the shell external fixation has motor III, is connected through hold-in range II between belt pulley III and the belt pulley II.
An electric cylinder II is arranged in the upper end face of the culture chamber, the electric cylinder II controls the blocking plate II to move up and down through a connecting rod, a feed inlet is formed in the upper end of the culture chamber, a culture medium discharge pipe is in through connection with a culture medium recovery box, and a culture medium adding pipe is in through connection with the culture medium box.
The lower end surface of the culture chamber is concaved downwards and is provided with a stem cell discharge cavity, an opening at the upper end of the stem cell discharge cavity is provided with a blocking plate III, an electric cylinder III is arranged below the blocking plate III, and the stem cell discharge cavity conveys stem cells to a stem cell collector through a stem cell discharge pipe.
The physiological saline water tank is used for adding physiological saline into the digestion chamber through the digestion chamber physiological saline adding pipe, and is communicated with the discharging opening through the centrifugal chamber physiological saline adding pipe.
The control system controls the opening and closing of all the electric components, and the power supply system provides power for all the electric components, which is needed by the work.
Preferably, be equipped with in the middle of the digestion chamber up end with shell fixed connection's middle piece, middle piece center be equipped with adipose tissue and add the mouth, adipose tissue adds mouthful to link up the digestion chamber inside with the shell outside and be connected, adipose tissue adds the mouth and seals through sealed lid, middle piece bottom surface is equipped with a plurality of annular gag lever posts I of arranging downwards, a plurality of gag lever posts I bottoms are connected with the bottom limiting plate jointly, a plurality of gag lever posts I intermediate slip is equipped with kickboard I, digestion chamber supernatant discharge pipe upper end wears to set up the shell outside and link up with the supernatant collecting box through the middle piece.
The feed opening is with digestion room and centrifugal chamber through connection, and the sealing disk rotates to set up in the change inslot portion, and sealing disk one side is equipped with the pivot, and the gear portion and the gear engagement of pivot lower extreme are connected, and motor I drive gear rotates.
The digestion chamber bottom surface indent downwards has the waste liquid discharge chamber, and waste liquid discharge chamber upper port passes through closure plate I and seals, and waste liquid discharge chamber below is equipped with electric jar I, and electric jar I controls the I reciprocates of closure plate, link up through the digestion chamber waste liquid discharge pipe between waste liquid discharge chamber and the waste liquid collecting box.
The digestion chamber lower end face is equipped with a plurality of annular gag lever post II of arranging downwards, and gag lever post II inserts to centrifugal chamber inside, and it is equipped with kickboard II to slide from top to bottom between a plurality of gag lever post II, and the centrifugal chamber supernatant discharge pipe other end wears to establish to the shell outside and link up with the supernatant collecting box through the digestion chamber bottom surface to be connected, and centrifugal chamber supernatant discharge pipe is equipped with a plurality of hole with kickboard II contact site.
The feed inlet internal diameter is greater than the II external diameters of closure plate, and the culture medium discharge pipe wears to establish to the shell outside and with culture medium recovery case through connection, and the culture medium adds the pipe and wears to establish to the shell outside and with culture medium case through connection, and the culture medium case adds new culture medium to digestion chamber inside through the culture medium and adds the pipe.
The digestive juice adding pipe, the digestive tract normal saline adding pipe, the centrifugal tract normal saline adding pipe, the digestive tract supernatant discharging pipe, the centrifugal tract supernatant discharging pipe, the digestive tract waste liquid discharging pipe, the culture medium adding pipe, the culture medium discharging pipe and the stem cell discharging pipe are all provided with liquid pumps.
Preferably, the gas protection system comprises a protection gas box, a gas pump and a carbon dioxide adding system, wherein a pressure sensor, an oxygen concentration sensor, a carbon dioxide sensor and an ultraviolet sterilization lamp II are arranged in the protection gas box.
The gas tank is connected with a gas release pipe in a penetrating way, the gas release pipe is provided with a solenoid valve I, the gas in the gas tank is conveyed to the digestion chamber by the gas pump through the gas supply pipe, a gas recovery pipe is connected between the digestion chamber and the gas tank in a penetrating way, and the gas recovery pipe is provided with a one-way valve.
The carbon dioxide adding system comprises a plurality of carbon dioxide cylinders, electromagnetic valves II are arranged on the air outlets of the carbon dioxide cylinders, the air outlets of the carbon dioxide cylinders are all in through connection with carbon dioxide adding pipes, and the carbon dioxide adding pipes are in through connection with the protective air box.
The pressure sensor, the oxygen concentration sensor, the carbon dioxide sensor, the ultraviolet sterilization lamp II, the air pump and the electromagnetic valve II are respectively and electrically connected with the control system.
Preferably, the gas protection system further comprises a nitrogen adding system, the nitrogen adding system comprises a plurality of nitrogen cylinders, electromagnetic valves III are arranged on the air outlets of the nitrogen cylinders, the air outlets of the nitrogen cylinders are all in through connection with a nitrogen adding pipe, the nitrogen adding pipe is in through connection with a protection gas tank, and the electromagnetic valves III are electrically connected with the control system.
Preferably, the circumference wall of the culture chamber is made of transparent materials, a plurality of constant temperature heaters and cameras are arranged at the positions corresponding to the culture chamber on the inner side of the shell, and the constant temperature heaters and the cameras are respectively and electrically connected with the control system.
Preferably, the digestion chamber inside be equipped with crushing agitating unit, crushing agitating unit include swivel, vertical pole, blade, a scraper blade, the swivel cover is established in the intermediate block outside, the protruding supporting disk that is equipped with of intermediate block lower extreme, the carousel sets up in the supporting disk top, vertical pole top passes through horizontal pole and swivel bottom surface fixed connection, vertical pole bottom and the inside bottom surface contact of digestion chamber, vertical pole side in the inside wall contact of digestion chamber, blade, scraper blade are fixed in vertical pole towards one side in digestion chamber center, scraper blade bottom surface and the inside bottom surface contact of digestion chamber, scraper blade length is greater than the inside radius of digestion chamber.
The top surface of the filter screen is flush with the bottom surface inside the digestion chamber.
The shell is externally fixed with a motor II, the motor II drives a belt pulley I to rotate, the belt pulley I drives conversion rotation through a synchronous belt I, and the motor II is electrically connected with a control system.
Preferably, a distributing ring is arranged below the culture chamber, the culture medium adding pipe is in through connection with the distributing ring, a collecting ring is arranged right above the distributing ring, and the collecting ring is in through connection with the culture medium discharging pipe.
A plurality of culture rods are uniformly distributed between the distribution ring and the collection ring, the upper ends of the culture rods are in through connection with the collection ring, and the lower ends of the culture rods are in through connection with the distribution ring.
The culture rod consists of an internal supporting framework and a filter screen wound outside the supporting framework, and nutrient substances in the culture medium can flow into the culture chamber through the filter screen.
Preferably, a digestive enzyme adding pipe is arranged in the culture chamber, and the tail end of the digestive enzyme adding pipe penetrates through the outer part of the shell and is communicated with the digestive enzyme box.
Preferably, the shell outside be equipped with the steady voltage box, steady voltage box inside is equipped with the steady voltage tube, steady voltage tube other end opening is located inside the feed opening, steady voltage box below opening part is equipped with static electret filter cotton, steady voltage box inside is equipped with sterilamp I, sterilamp I and sterilamp I electric connection.
Preferably, the blanking opening is in a truncated cone shape with a narrow upper part and a wide lower part.
Compared with the prior art, the invention has the beneficial effects that:
(1) The digestion chamber, the centrifugal chamber and the culture chamber are integrated inside the shell from top to bottom, so that the integration level of the stem cell culture process device is improved, the whole volume of equipment required by stem cell culture is effectively reduced, and the space utilization rate of a workshop is improved.
(2) The unmanned dry running culture process is realized, the influence of external bacteria on the dry running culture process is reduced, and a sterile environment for the dry running culture is better created.
Drawings
The invention will be further described with reference to the drawings and examples.
FIG. 1 is a flow chart of a stem cell preparation system according to the present invention,
FIG. 2 is an external view showing a stem cell preparation apparatus according to the present invention,
FIG. 3 is a view showing the construction of the inside of the stem cell preparation apparatus according to the present invention after the housing is removed,
FIG. 4 is a cross-sectional view of A-A surface of a stem cell preparation apparatus according to the present invention,
FIG. 5 is a B-B side sectional view of a stem cell preparation apparatus of the present invention,
FIG. 6 is a C-C side sectional view of a stem cell preparation apparatus of the present invention,
FIG. 7 is a partially enlarged view of the stem cell preparation apparatus D according to the present invention,
FIG. 8 is a partial enlarged view of the stem cell preparation apparatus E according to the present invention,
FIG. 9 is a schematic diagram showing the driving of the pulverizing and stirring device of the stem cell preparation apparatus of the present invention,
FIG. 10 is a schematic diagram of a sealing disk drive of a stem cell preparation apparatus according to the present invention,
FIG. 11 is a schematic diagram showing the driving of a centrifugal chamber of a stem cell preparation apparatus according to the present invention,
figure 12 is a block diagram of a shielding gas system of the present invention,
figure 13 is a cross-sectional view of the gas box of the shielding gas system of the present invention,
FIG. 14 is a partial enlarged view of the culture apparatus according to the invention.
In the figure: 100-stem cell preparation device, 200-control system, 300-power supply system, 400-digestive juice tank, 500-physiological saline tank, 600-supernatant collecting tank, 700-waste liquid collecting tank, 800-culture medium tank, 900-culture medium recovery tank, 1000-stem cell collector and 1100-digestive enzyme tank;
01-digestive juice adding tube, 021-digestive tract physiological saline adding tube, 022-centrifugal tract physiological saline adding tube, 031-digestive tract supernatant discharging tube, 032-centrifugal tract supernatant discharging tube, 04-digestive tract waste liquid discharging tube, 051-protective gas supplying tube, 052-protective gas recovering tube, 061-culture medium adding tube, 062-culture medium discharging tube, 07-stem cell discharging tube, 08-digestive enzyme adding tube;
1-shell, 1 a-constant temperature heater and 1 b-camera;
2-middle block, 2 A-Adipose tissue adding port, 2 b-supporting disk, 2 c-sealing cover, 2 d-limiting rod I, 2 e-bottom limiting plate and 2 f-floating plate I;
the device comprises a 3-digestion chamber, a 3 a-feed opening, a 3 b-filter screen, a 3 c-rotary groove, a 3 d-sealing disc, a 3d 1-rotary shaft, a 3 e-motor I, a 3e 1-gear, a 3 f-waste liquid discharge cavity, a 3 g-electric cylinder I, a 3g 1-blocking plate I and a 3 h-limiting rod II;
4-crushing stirring device, 4 a-swivel, 4 b-vertical rod, 4 c-blade and 4 d-scraper;
5-motor II, 5 a-belt pulley I, 5 b-synchronous belt I;
6-centrifugal chambers, 6 a-stirring plates, 6 b-floating plates II, 6 c-belt pulleys II, 6 d-blocking plates II, 6d 1-connecting rods and 6 e-electric cylinders II;
7-motor III, 7 a-belt pulley III, 7 b-synchronous belt II;
8-culture chamber, 8 a-feed inlet, 8 b-distribution ring, 8 c-culture rod, 8c 1-collection ring, 8 d-stem cell discharge cavity, 8 e-closure plate III, 8 f-electric cylinder III;
9-voltage stabilizing box, 9 a-voltage stabilizing tube, 9 b-ultraviolet sterilizing lamp I, 9 c-electrostatic electret filter cotton;
10-gas protection system, 10 a-protection gas tank, 10 b-pressure sensor, 10 c-oxygen concentration sensor, 10 d-carbon dioxide concentration sensor, 10 e-bleeder, 10e 1-solenoid valve I, 10 f-air pump, 10 g-check valve, 10 h-carbon dioxide addition system, 10h 1-carbon dioxide gas cylinder, 10h 2-solenoid valve II, 10h 3-carbon dioxide addition pipe, 10 i-nitrogen addition system, 10i 1-nitrogen gas cylinder, 10i 2-solenoid valve III, 10i 3-nitrogen addition pipe, 10 j-ultraviolet sterilization lamp II.
Detailed Description
The drawings are preferred embodiments of the stem cell preparation system, and the present invention will be described in further detail with reference to the accompanying drawings.
Referring to fig. 1, a stem cell preparation system includes a stem cell preparation apparatus 100, a control system 200, a power supply system 300, a digestive juice tank 400, a physiological saline tank 500, a supernatant collecting tank 600, a waste liquid collecting tank 700, a culture medium tank 800, a culture medium recovery tank 900, a stem cell collector 1000, a digestive enzyme tank 1100, and a gas protection system 10.
Referring to FIGS. 2 to 5, the stem cell preparation apparatus 100 includes a housing 1, a digestion chamber 3, a centrifugal chamber 6, and a culture chamber 8, which are sequentially connected from top to bottom, inside the housing 1.
As shown in fig. 8, a middle block 2 fixedly connected with the housing 1 is provided in the middle of the upper end surface of the digestion chamber 3, an adipose tissue adding port 2a is provided in the center of the middle block 2, the adipose tissue adding port 2a connects the inside of the digestion chamber 3 with the outside of the housing 1, and the digestion liquid tank 400 is connected with the adipose tissue adding port 2a through the digestion liquid adding pipe 01. The adipose tissue-adding port 2a is sealed by a sealing cap 2 c.
The middle block 2 bottom surface is equipped with a plurality of gag lever posts I2 d that annular was arranged downwards, and a plurality of gag lever posts I2 d bottoms are connected with bottom limiting plate 2e jointly, and the slip is equipped with kickboard I2 f in the middle of a plurality of gag lever posts I2 d, and kickboard I2 f upper end fixedly connected with digestion chamber supernatant discharge pipe 031, and digestion chamber supernatant discharge pipe 031 and kickboard I2 f contact position open has the hole, and digestion chamber supernatant discharge pipe 031 upper end wears to set up shell 1 outside and link up with supernatant collecting box 600 through middle block 2.
The fat tissue is placed into the digestion chamber 3 through the fat tissue addition port 2a, and the digestive juice is added into the digestion chamber 3 through the digestive juice addition tube 01, so that the fat tissue is digested and separated into a precipitate and a supernatant. The density of the float plate i 2f is greater than that of the supernatant and less than that of the sediment, so the float plate i 2f is located between the sediment and the supernatant. The liquid pump on the digestion chamber supernatant discharge pipe 031 is turned on to discharge the whole supernatant.
A feed opening 3a is arranged in the middle of the lower end surface of the digestion chamber 3, and the feed opening 3a is in a truncated cone shape with a narrow upper part and a wide lower part. The digestion chamber 3 is communicated with the centrifugal chamber 6 through the feed opening 3a, a filter screen 3b is arranged at the upper end of the feed opening 3a, and a sealing disc 3d is arranged at the lower end of the filter screen 3 b. As shown in fig. 6, 7 and 10, the sealing disc 3d is rotatably disposed in the rotary groove 3c, a rotary shaft 3d1 is disposed on one side of the sealing disc 3d, a gear portion at the lower end of the rotary shaft 3d1 is engaged with the gear 3e1, and the motor i 3e drives the gear 3e1 to rotate.
The motor I3 e is started, and the sealing disc 3d is driven to rotate in the rotary groove 3c through the gear 3e1, so that the digestion chamber 3 is in through connection with the centrifugal chamber 6. The precipitate in the digestion chamber 3 flows into the centrifugal chamber 6 after being filtered by the filter screen 3 b.
The bottom surface of the digestion chamber 3 is provided with a waste liquid discharge cavity 3f in a downward concave manner, an upper port of the waste liquid discharge cavity 3f is sealed through a blocking plate I3 g1, an electric cylinder I3 g is arranged below the waste liquid discharge cavity 3f, the electric cylinder I3 g controls the blocking plate I3 g1 to move up and down, and the waste liquid discharge cavity 3f is in through connection with the waste liquid collecting box 700 through a digestion chamber waste liquid discharge pipe 04.
The physiological saline tank 500 adds physiological saline to the digestion chamber 3 through the digestion chamber physiological saline addition pipe 021, and the physiological saline washes the substances remaining above the filter screen 3b into the waste liquid discharge chamber 3f, and then discharges the substances into the waste liquid collection tank 700 through the digestion chamber waste liquid discharge pipe 04 for concentrated treatment.
As shown in fig. 9, a crushing and stirring device 4 is arranged in the digestion chamber 3, and the crushing and stirring device 4 comprises a swivel 4a, a vertical rod 4b, a blade 4c and a scraper 4d. The swivel 4a is sleeved outside the middle block 2, the lower end of the middle block 2 is convexly provided with a supporting disc 2b, and the swivel 4a is arranged above the supporting disc 2 b. The top surface of the vertical rod 4b is fixedly connected with the bottom surface of the swivel 4a through a horizontal rod, the bottom of the vertical rod 4b is contacted with the bottom surface inside the digestion chamber 3, and the side surface of the vertical rod 4b is contacted with the inner wall of the digestion chamber 3. The blade 4c and the scraper 4d are fixed on one side of the vertical rod 4b facing the center of the digestion chamber 3, the bottom surface of the scraper 4d is contacted with the inner bottom surface of the digestion chamber 3, and the length of the scraper 4b is larger than the inner radius of the digestion chamber 3. The blade 4c chops the adipose tissue inside the digestion chamber 3.
The top surface of the filter screen 3b is flush with the inner bottom surface of the digestion chamber 3.
The shell 1 is externally fixed with a motor II 5, the motor II 5 drives a belt pulley I5 a to rotate, the belt pulley I5 a drives a conversion 4a to rotate through a synchronous belt I5 b, and the motor II 5 is electrically connected with the control system 200.
The lower end face of the digestion chamber 3 is downwards provided with a plurality of limiting rods II 3h which are annularly arranged, the limiting rods II 3h are inserted into the centrifugal chamber 6, and a floating plate II 6b is arranged between the limiting rods II 3h in a sliding mode. The upper end face of the floating plate II 6b is fixedly connected with one end of a supernatant discharge pipe 032 of the centrifugal chamber, the other end of the supernatant discharge pipe 032 of the centrifugal chamber passes through the bottom face of the digestion chamber 3, penetrates the outer part of the shell 1 and is in through connection with the supernatant collecting box 600, and a plurality of holes are formed in the contact part of the supernatant discharge pipe 032 of the centrifugal chamber and the floating plate II 6b.
The inner circumference surface of the centrifugal chamber 6 is convexly provided with a plurality of stirring plates 6a, and the opening of the bottom surface of the centrifugal chamber 6 is sealed by a circular blocking plate II 6 d. As shown in figure 11, a belt pulley II 6c is sleeved outside the centrifugal chamber 6, a motor III 7 is fixed outside the shell 1, the motor III 7 drives a belt pulley III 7a to rotate, and the belt pulley III 7a is connected with the belt pulley II 6c through a synchronous belt II 7 b.
The physiological saline tank 500 is connected to the discharge port 3a through the physiological saline addition pipe 022, and the physiological saline is injected into the centrifugal chamber 6 to be mixed with the substance discharged from the digestion chamber 3. The motor III 7 drives the centrifugal chamber 6 to rotate, so that the stirring plate 6a drives the liquid in the centrifugal chamber 6 to rotate, centrifugal force is generated, and the sediment is subjected to centrifugal treatment, so that the sediment is separated into supernatant fluid and stem cells below the supernatant fluid. The floating plate II 6b is positioned between the supernatant and the stem cells, and the supernatant is discharged into the supernatant collecting tank 600 by starting the liquid pump on the supernatant discharge pipe 032 of the centrifugal chamber.
An electric cylinder II 6e is arranged in the upper end face of the culture chamber 8, and the electric cylinder II 6e controls the blocking plate II 6d to move up and down through a connecting rod 6d 1. When the blocking plate II 6d moves up to the highest point, the opening on the bottom surface of the centrifugal chamber 6 can be blocked, the blocking plate II 6d moves down, and the dry matter inside the centrifugal chamber 6 runs into the culture chamber 8. The upper end of the culture chamber 8 is provided with a feed inlet 8a, the inner diameter of the feed inlet 8a is larger than the outer diameter of a blocking plate II 6d, and the blocking plate II 6d is conical, so that the dry running inside the centrifugal chamber 6 is facilitated to flow into the culture chamber 8.
As shown in FIG. 14, a distribution ring 8b is provided below the inside of the culture chamber 8, a medium addition pipe 061 is connected to the distribution ring 8b, a collection ring 8c1 is provided right above the distribution ring 8b, and the collection ring 8c1 is connected to a medium discharge pipe 062.
A plurality of culture rods 8c are uniformly distributed between the distribution ring 8b and the collection ring 8c1, the upper ends of the culture rods 8c are in through connection with the collection ring 8c1, and the lower ends of the culture rods 8c are in through connection with the distribution ring 8 b.
The culture rod 8c consists of an inner supporting framework and a filter screen wound outside the supporting framework, and nutrient substances in the culture medium can flow into the culture chamber through the filter screen. The dry running from the centrifugal chamber 6 flows onto the culture rod 8c, the aperture of the filter screen is smaller than the dry running diameter, and meanwhile, the filter screen has certain friction force, so that the dry running is convenient to hang on the culture rod 8 c. In order to detach the stem cells from the culture rod 8c after the stem cell culture, a digestive enzyme adding tube 08 is provided in the culture chamber 8, and the end of the digestive enzyme adding tube 08 is inserted outside the housing 1 and is connected to the digestive enzyme box 1100. The liquid pump on the digestive enzyme adding tube 08 injects digestive enzymes into the culture chamber 8 to separate the dry run from the culture rod 8 c.
The medium discharge pipe 062 is penetrating to the outside of the casing 1 and is connected to the medium recovery tank 900, the medium addition pipe 061 is penetrating to the outside of the casing 1 and is connected to the medium tank 800, and the medium tank 800 adds a new medium to the inside of the distribution ring 8b of the digestion chamber 8 through the medium addition pipe 061.
The lower end surface of the culture chamber 8 is provided with a stem cell discharge cavity 8d which is concave downwards, an opening at the upper end of the stem cell discharge cavity 8d is sealed by a plugging plate III 8e, an electric cylinder III 8f arranged below the stem cell discharge cavity 8d controls the plugging plate III 8e to move up and down, and the stem cell discharge cavity 8d conveys stem cells to the stem cell collector 1000 through a stem cell discharge pipe 07.
The circumference wall of the culture chamber 8 is made of transparent materials, a plurality of constant temperature heaters 1a and cameras 1b are arranged at the positions corresponding to the culture chamber 8 on the inner side of the shell 1, and the constant temperature heaters 1a and the cameras 1b are respectively and electrically connected with the control system 200. The constant temperature heater 1a ensures that the inside of the culture chamber 8 is in a constant temperature state, the camera 1b transmits the influence to the control center in real time, and a technician observes the dry running culture degree inside the culture chamber 8 through an image.
The digestive juice adding pipe 01, the digestive tract normal saline adding pipe 021, the centrifugal tract normal saline adding pipe 022, the digestive tract supernatant discharging pipe 031, the centrifugal tract supernatant discharging pipe 032, the digestive tract waste liquid discharging pipe 04, the culture medium adding pipe 061, the culture medium discharging pipe 062, the stem cell discharging pipe 07 and the digestive enzyme adding pipe 08 are all provided with liquid extracting pumps.
The control system 200 controls the turning on and off of the electrical components, and the power supply system 300 provides the power required for the operation of the electrical components.
As shown in fig. 12 and 13, the gas protection system 10 includes a protection gas tank 10a, a gas pump 10f, and a carbon dioxide addition system 10h, and a pressure sensor 10b, an oxygen concentration sensor 10c, a carbon dioxide sensor 10d, and an ultraviolet germicidal lamp ii 10j are provided inside the protection gas tank 10 a.
The protective gas box 10a is connected with a gas release pipe 10e in a penetrating way, the gas release pipe 10e is provided with a solenoid valve I10 e1, the gas pump 10f conveys gas in the protective gas box 10a into the digestion chamber 8 through the protective gas supply pipe 051, a protective gas recovery pipe 052 is connected between the digestion chamber 8 and the protective gas box 10a in a penetrating way, and the protective gas recovery pipe 052 is provided with a one-way valve 10g.
The carbon dioxide adding system 10h comprises a plurality of carbon dioxide gas cylinders 10h1, electromagnetic valves II 10h2 are arranged on the gas outlets of the carbon dioxide gas cylinders 10h1, the gas outlets of the carbon dioxide gas cylinders 10h1 are all in through connection with a carbon dioxide adding pipe 10h3, and the carbon dioxide adding pipe 10h3 is in through connection with a protective gas box 10 a.
The pressure sensor 10b, the oxygen concentration sensor 10c, the carbon dioxide sensor 10d, the ultraviolet germicidal lamp ii 10j, the air pump 10f, and the electromagnetic valve ii 10h2 are electrically connected to the control system 200, respectively.
The gas protection system 10 further comprises a nitrogen adding system 10i, the nitrogen adding system 10i comprises a plurality of nitrogen cylinders 10i1, electromagnetic valves III 10i2 are arranged on the air outlets of the nitrogen cylinders 10i1, the air outlets of the plurality of nitrogen cylinders 10i1 are all in through connection with a nitrogen adding pipe 10i3, the nitrogen adding pipe 10i3 is in through connection with a protection gas box 10a, and the electromagnetic valves III 10i2 are electrically connected with the control system 200. The nitrogen addition system 10i can adjust the oxygen concentration inside the shielding gas.
The shell 1 outside be equipped with steady voltage box 9, steady voltage box 9 inside is equipped with steady voltage tube 9a, steady voltage tube 9a other end opening is located inside feed opening 3a, steady voltage box 9 below opening part is equipped with static electret filter cotton 9c, steady voltage box 9 inside is equipped with sterilamp I9 b, sterilamp I9 b and sterilamp I9 b electric connection.
The embodiments of the present invention have been described in detail with reference to the drawings, but the present invention is not limited to the above embodiments, and various changes can be made within the knowledge of one of ordinary skill in the art without departing from the spirit of the present invention.
Claims (10)
1. A stem cell preparation system, characterized in that:
comprises a stem cell preparation device (100), a control system (200), a power supply system (300), a digestive juice box (400), a physiological saline box (500), a supernatant collecting box (600), a waste liquid collecting box (700), a culture medium box (800), a culture medium recycling box (900) and a stem cell collector (1000),
the stem cell preparation device (100) comprises a shell (1), a digestion chamber (3), a centrifugal chamber (6) and a culture chamber (8) which are sequentially connected from top to bottom and are positioned in the shell (1),
the upper end surface of the digestion chamber (3) is provided with an adipose tissue adding port (2 a), the digestion liquid tank (400) is in through connection with the adipose tissue adding port (2 a) through a digestion liquid adding pipe (01), a floating plate I (2 f) is arranged in the digestion chamber (3), the upper end of the floating plate I (2 f) is fixedly connected with a digestion chamber supernatant discharge pipe (031), a hole is arranged at the contact part of the digestion chamber supernatant discharge pipe (031) and the floating plate I (2 f), the digestion chamber supernatant discharge pipe (031) is in through connection with a supernatant collecting box (600),
a feed opening (3 a) is arranged in the middle of the lower end surface of the digestion chamber (3), a filter screen (3 b) is arranged at the upper end of the feed opening (3 a), a sealing disc (3 d) is arranged at the lower end of the filter screen (3 b),
a floating plate II (6 b) is arranged in the centrifugal chamber (6), the upper end surface of the floating plate II (6 b) is fixedly connected with one end of a supernatant discharge pipe (032) of the centrifugal chamber, the supernatant discharge pipe (032) of the centrifugal chamber is in through connection with a supernatant collecting box (600), a plurality of stirring plates (6 a) are convexly arranged on the circumferential surface in the centrifugal chamber (6), the bottom surface opening of the centrifugal chamber (6) is sealed by a circular blocking plate II (6 d), a belt pulley II (6 c) is sleeved outside the centrifugal chamber (6), a motor III (7) is fixed outside the casing (1), the belt pulley III (7 a) is connected with the belt pulley II (6 c) by a synchronous belt II (7 b),
an electric cylinder II (6 e) is arranged in the upper end surface of the culture chamber (8), the electric cylinder II (6 e) controls the blocking plate II (6 d) to move up and down through a connecting rod (6 d 1), a feed inlet (8 a) is arranged at the upper end of the culture chamber (8), a culture medium discharge pipe (062) is connected with a culture medium recovery box (900) in a penetrating way, a culture medium adding pipe (061) is connected with the culture medium box (800) in a penetrating way,
a stem cell discharge cavity (8 d) is concaved downwards from the lower end surface of the culture chamber (8), a closure plate III (8 e) is arranged at the upper end opening of the stem cell discharge cavity (8 d), an electric cylinder III (8 f) is arranged below the closure plate III (8 e), the stem cell discharge cavity (8 d) conveys stem cells to a stem cell collector (1000) through a stem cell discharge pipe (07),
the physiological saline tank (500) is used for adding physiological saline into the digestion chamber (3) through a physiological saline adding pipe (021) of the digestion chamber and is communicated with the discharging opening (3 a) through a physiological saline adding pipe (022) of the centrifugal chamber,
the control system (200) controls the opening and closing of the electric components, and the power supply system (300) provides power required by the work for the electric components.
2. A stem cell preparation system according to claim 1, wherein:
an intermediate block (2) fixedly connected with the shell (1) is arranged in the middle of the upper end face of the digestion chamber (3), an adipose tissue adding port (2 a) is arranged in the center of the intermediate block (2), the adipose tissue adding port (2 a) is used for connecting the inside of the digestion chamber (3) with the outside of the shell (1) in a penetrating way, the adipose tissue adding port (2 a) is sealed by a sealing cover (2 c), a plurality of limit rods I (2 d) which are annularly arranged are downwards arranged on the bottom face of the intermediate block (2), the bottoms of the limit rods I (2 d) are jointly connected with a bottom limit plate (2 e), a floating plate I (2 f) is slidably arranged in the middle of the limit rods I (2 d), the upper end of a supernatant discharge pipe (031) of the digestion chamber penetrates through the intermediate block (2) to be arranged outside the shell (1) and is connected with a supernatant collecting box (600) in a penetrating way,
the digestion chamber (3) is communicated with the centrifugal chamber (6) through the blanking opening (3 a), the sealing disc (3 d) is rotationally arranged in the rotary groove (3 c), a rotary shaft (3 d 1) is arranged on one side of the sealing disc (3 d), a gear part at the lower end of the rotary shaft (3 d 1) is meshed with the gear (3 e 1), the motor I (3 e) drives the gear (3 e 1) to rotate,
a waste liquid discharge cavity (3 f) is concaved downwards and inwards on the bottom surface of the digestion chamber (3), the upper port of the waste liquid discharge cavity (3 f) is sealed by a blocking plate I (3 g 1), an electric cylinder I (3 g) is arranged below the waste liquid discharge cavity (3 f), the electric cylinder I (3 g) controls the blocking plate I (3 g 1) to move up and down, the waste liquid discharge cavity (3 f) is communicated with a waste liquid collecting box (700) through a digestion chamber waste liquid discharge pipe (04),
a plurality of limit rods II (3 h) which are annularly arranged are downwards arranged on the lower end surface of the digestion chamber (3), the limit rods II (3 h) are inserted into the centrifugal chamber (6), a floating plate II (6 b) is arranged between the limit rods II (3 h) in a sliding way up and down, the other end of a supernatant discharge pipe (032) of the centrifugal chamber penetrates through the bottom surface of the digestion chamber (3) to the outside of the shell (1) and is in through connection with a supernatant collecting box (600), a plurality of holes are arranged at the contact part of the supernatant discharge pipe (032) of the centrifugal chamber and the floating plate II (6 b),
the inner diameter of the feed port (8 a) is larger than the outer diameter of the blocking plate II (6 d), the culture medium discharge pipe (062) is penetrated outside the shell (1) and is in through connection with the culture medium recovery tank (900), the culture medium adding pipe (061) is penetrated outside the shell (1) and is in through connection with the culture medium tank (800), the culture medium tank (800) adds new culture medium into the digestion chamber (8) through the culture medium adding pipe (061),
the digestive juice adding pipe (01), the digestive tract normal saline adding pipe (021), the centrifugal tract normal saline adding pipe (022), the digestive tract supernatant discharging pipe (031), the centrifugal tract supernatant discharging pipe (032), the digestive tract waste liquid discharging pipe (04), the culture medium adding pipe (061), the culture medium discharging pipe (062) and the stem cell discharging pipe (07) are all provided with liquid pumps.
3. A stem cell preparation system according to claim 2, wherein:
the gas protection system (10) is further included, the gas protection system (10) comprises a protection gas box (10 a), a gas pump (10 f) and a carbon dioxide adding system (10 h), a pressure sensor (10 b), an oxygen concentration sensor (10 c), a carbon dioxide sensor (10 d) and an ultraviolet sterilization lamp II (10 j) are arranged inside the protection gas box (10 a),
the protective gas box (10 a) is connected with a gas release pipe (10 e) in a penetrating way, the gas release pipe (10 e) is provided with a solenoid valve I (10 e 1), the gas pump (10 f) conveys gas in the protective gas box (10 a) to the inside of the digestion chamber (8) through a protective gas supply pipe (051), a protective gas recovery pipe (052) is connected between the digestion chamber (8) and the protective gas box (10 a) in a penetrating way, a one-way valve is arranged on the protective gas recovery pipe (052),
the carbon dioxide adding system (10 h) comprises a plurality of carbon dioxide gas cylinders (10 h 1), electromagnetic valves II (10 h 2) are arranged on the gas outlets of the carbon dioxide gas cylinders (10 h 1), the gas outlets of the carbon dioxide gas cylinders (10 h 1) are all in through connection with a carbon dioxide adding pipe (10 h 3), the carbon dioxide adding pipe (10 h 3) is in through connection with a protective gas box (10 a),
the pressure sensor (10 b), the oxygen concentration sensor (10 c), the carbon dioxide sensor (10 d), the ultraviolet sterilization lamp II (10 j), the air pump (10 f) and the electromagnetic valve II (10 h 2) are respectively and electrically connected with the control system (200).
4. A stem cell preparation system according to claim 3, wherein:
the gas protection system (10) further comprises a nitrogen adding system (10 i), the nitrogen adding system (10 i) comprises a plurality of nitrogen cylinders (10 i 1), electromagnetic valves III (10 i 2) are arranged on the air outlets of the nitrogen cylinders (10 i 1), the air outlets of the plurality of nitrogen cylinders (10 i 1) are all connected with a nitrogen adding pipe (10 i 3) in a penetrating mode, the nitrogen adding pipe (10 i 3) is connected with a protection gas tank (10 a) in a penetrating mode, and the electromagnetic valves III (10 i 2) are electrically connected with the control system (200).
5. A stem cell preparation system according to claim 2, wherein:
the circumference wall of the culture chamber (8) is made of transparent materials, a plurality of constant temperature heaters (1 a) and cameras (1 b) are arranged at the positions corresponding to the culture chamber (8) on the inner side of the shell (1), and the constant temperature heaters (1 a) and the cameras (1 b) are respectively and electrically connected with the control system (200).
6. A stem cell preparation system according to claim 2, wherein:
the inside of the digestion chamber (3) is provided with a crushing and stirring device (4), the crushing and stirring device (4) comprises a swivel (4 a), a vertical rod (4 b), a blade (4 c) and a scraping plate (4 d), the swivel (4 a) is sleeved outside the middle block (2), the lower end of the middle block (2) is convexly provided with a supporting disc (2 b), the rotating disc (4 a) is arranged above the supporting disc (2 b), the top surface of the vertical rod (4 b) is fixedly connected with the bottom surface of the swivel (4 a) through a horizontal rod, the bottom of the vertical rod (4 b) is contacted with the inner bottom surface of the digestion chamber (3), the side surface of the vertical rod (4 b) is contacted with the inner wall of the digestion chamber (3), the blade (4 c) and the scraping plate (4 d) are fixed on one side of the vertical rod (4 b) facing the center of the digestion chamber (3), the bottom surface of the scraping plate (4 d) is contacted with the inner bottom surface of the digestion chamber (3), the length of the scraping plate (4 b) is larger than the inner radius of the digestion chamber (3),
the top surface of the filter screen (3 b) is flush with the bottom surface of the digestion chamber (3),
a motor II (5) is fixed outside the shell (1), the motor II (5) drives a belt pulley I (5 a) to rotate, the belt pulley I (5 a) drives a conversion (4 a) to rotate through a synchronous belt I (5 b), and the motor II (5) is electrically connected with a control system (200).
7. A stem cell preparation system according to claim 2, wherein:
a distribution ring (8 b) is arranged below the inner part of the culture chamber (8), a culture medium adding pipe (061) is communicated with the distribution ring (8 b), a collection ring (8 c 1) is arranged right above the distribution ring (8 b), the collection ring (8 c 1) is communicated with a culture medium discharging pipe (062),
a plurality of culture rods (8 c) are uniformly distributed between the distribution ring (8 b) and the collection ring (8 c 1), the upper ends of the culture rods (8 c) are communicated with the collection ring (8 c 1), the lower ends of the culture rods (8 c) are communicated with the distribution ring (8 b),
the culture rod (8 c) consists of an inner supporting framework and a filter screen wound outside the supporting framework, and nutrient substances in the culture medium can flow into the culture chamber through the filter screen.
8. The stem cell preparation system of claim 7, wherein:
the inside of the culture chamber (8) is provided with a digestive enzyme adding pipe (08), and the tail end of the digestive enzyme adding pipe (08) is penetrated to the outside of the shell (1) and is communicated with the digestive enzyme box (1100).
9. A stem cell preparation system according to claim 2, wherein:
the shell (1) outside be equipped with steady voltage box (9), steady voltage box (9) inside is equipped with steady voltage tube (9 a), steady voltage tube (9 a) other end opening is located inside feed opening (3 a), steady voltage box (9) below opening part is equipped with static electret filter cotton (9 c), steady voltage box (9) inside is equipped with sterilamp I (9 b), sterilamp I (9 b) and sterilamp I (9 b) electric connection.
10. A stem cell preparation system according to claim 2, wherein:
the blanking opening (3 a) is in a truncated cone shape with a narrow upper part and a wide lower part.
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| CN113862118A (en) * | 2021-10-27 | 2021-12-31 | 山东大学齐鲁医院 | Tumor cell separation device |
| CN117987242B (en) * | 2024-04-03 | 2024-07-09 | 苏州赛普生物科技股份有限公司 | SVF cell separation device and SVF cell separation method |
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| CN101041692A (en) * | 2006-10-20 | 2007-09-26 | 沈阳药科大学 | Scorpion analgesic antibacterial active peptide and preparation method thereof |
| JP2010246446A (en) * | 2009-04-14 | 2010-11-04 | Olympus Corp | Device for treating adipose tissue and apparatus for separating cell |
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