CN112680356A

CN112680356A - Stem cell directional differentiation device for cell culture and use method thereof

Info

Publication number: CN112680356A
Application number: CN202110099227.6A
Authority: CN
Inventors: 李宁
Original assignee: Individual
Current assignee: Individual
Priority date: 2021-01-25
Filing date: 2021-01-25
Publication date: 2021-04-20

Abstract

The invention provides a stem cell directional differentiation device for cell culture, which comprises a box body, a cover plate, a culture dish and an antibacterial box, wherein the opening at the top of the box body is movably provided with the detachable cover plate, the center of the top of the cover plate is fixedly connected with the antibacterial box, the lower end of the antibacterial box extends into the box body, the front side and the rear side in the box body are respectively and fixedly connected with connecting blocks, a culture dish capable of horizontally rotating is movably connected between the connecting blocks, the left side and the right side of the lower end in the box body are respectively and rotatably connected with screw rods, the screw rods are synchronously linked through a chain wheel and a chain, a positive and negative motor and the screw rods are synchronously linked through the chain wheel and the chain, a bearing plate capable of lifting up and down is in threaded connection between the two upper ends of a partition plate, and the top, the stem cell suspension liquid drop is easy to be polluted by bacteria when being added, and the prior culture dish is inconvenient for real-time observation.

Description

Stem cell directional differentiation device for cell culture and use method thereof

Technical Field

The invention relates to the technical field of cell culture, in particular to a stem cell directional differentiation device for cell culture and a using method thereof.

Background

Heart failure caused by myocardial cell dysfunction is a major disease worldwide, and the cause of eventual death in heart failure patients is mainly loss of cardiac pumping function or arrhythmia. The annual death rate of severe heart failure patients exceeds 50%, although late-stage patients can be transplanted with heart, due to the lack of donor organs, about 20% of patients die while waiting for organ transplantation, due to the high morbidity and mortality of heart failure, the death of transplanted heart, complications such as immunological rejection and the like, and the objective situation of late-stage failure of transplanted heart, the development of new treatment methods for improving the functions of cardiomyocytes and preventing the occurrence of heart failure is urgently needed, the current treatment methods for heart failure mainly focus on early-stage vascular regeneration and inhibiting further cardiomyocyte loss, the cardiomyocytes are terminally differentiated cells, the proliferation capacity is extremely limited, the recovery after the loss is extremely difficult, the cardiomyocyte transplantation therapy focuses on reconstructing the myocardial function and recovering the pumping capacity of regenerated myocardial tissue, and researches show that the cardiomyocytes injected into the heart can form new myocardial tissue, however, the key factor of cell transplantation therapy is to determine a suitable donor cell type, and therefore, how to induce the stem cells to directionally differentiate into the myocardial cells for medical treatment is an urgent technical problem to be solved.

At present, in the process of cell culture differentiation, the stem cell suspension liquid drops are easy to be polluted by bacteria when added, and meanwhile, the existing culture dish is inconvenient for real-time observation, so that the problem is solved by providing a stem cell directional differentiation device for cell culture.

Disclosure of Invention

Technical problem to be solved

The invention aims to solve the problems that in the cell culture and differentiation process in the prior art, suspension liquid drops of stem cells are easily polluted by bacteria when added, and meanwhile, the existing culture dish is inconvenient for real-time observation.

(II) technical scheme

The invention relates to a stem cell directional differentiation device for cell culture and an application method thereof, and aims and effects are achieved by the following specific technical means: a stem cell directional differentiation device for cell culture and a using method thereof comprise a box body, a cover plate, a culture dish and an antibacterial box, wherein the opening at the top of the box body is movably provided with a detachable cover plate, the center of the top of the cover plate is fixedly connected with the antibacterial box, the lower end of the antibacterial box extends into the box body, the front side and the rear side inside the box body are both fixedly connected with connecting blocks, the culture dish capable of horizontally rotating is movably connected between the connecting blocks, the lower end inside the box body is fixedly connected with a first partition plate, the upper end inside the box body is fixedly connected with a second partition plate, a singlechip controller is fixedly arranged between the first partition plate and the second partition plate, a positive and negative motor is fixedly arranged between the first partition plate and the second partition plate, the left side and the right side of the lower end inside the box body are both rotatably connected with lead screws, the lead screws are, the top of the bearing plate is fixedly provided with a CCD detection module.

Preferably, two top right sides central point of baffle put to rotate and are connected with vertical driven shaft, the driven shaft top extends to loading board upper end outside fixedly connected with drive gear, the outer ring gear of culture dish lower extreme outside fixedly connected with, drive gear and outer ring gear meshing joint, culture dish top outside fixedly connected with annular buckle, the shifting chute runs through the groove for the arc, the inside sliding connection of shifting chute is corresponded to annular buckle, the apron bottom is embedded to have the universal ball of round, universal ball and annular buckle top sliding connection.

Preferably, the box top is equipped with square form draw-in groove, and the apron corresponds the inside hollow position joint of draw-in groove, the inside equal fixedly connected with air cushion of intercommunication of both sides of draw-in groove.

Preferably, the fungi-proofing box comprises an auxiliary culture cover, an inlet, a sliding chute, a sliding rod and side fixing plates, wherein the center of the top of the fungi-proofing box is provided with the inlet, the bottom of the fungi-proofing box is provided with the auxiliary culture cover which is connected in a rotating manner, the inside of the fungi-proofing box is fixedly connected with a positive and negative motor, the output shaft of the positive and negative motor is connected with the central shaft of the auxiliary culture cover through a bevel gear in a transmission manner, the left side and the right side of the rear side of the inside of the fungi-proofing box, which are positioned at the central position of the lower end of the inlet, are respectively provided with an arc-shaped sliding chute, the left side and the right side of the inlet are respectively and rotatably connected with a clamping air injection assembly for closing the inlet, the left side and the right side fixing plates are respectively and vertically and fixedly connected with the top of the, the rear side of the interior of the fungi-proofing box is provided with a plurality of air suction holes in sequence at equal intervals, the rear side of the fungi-proofing box is fixedly provided with an air suction pump, and an air suction pipe is fixedly arranged between the air suction pump and the air suction holes.

Preferably, the jet-propelled subassembly of centre gripping is including the grip block, the clamp ring subassembly, tangent plane and venthole, the grip block is close to import center one side and is equipped with the arc wall, be equipped with a plurality of ventholes in the arc wall, the grip block bottom is close to import center one side and establishes to the tangent plane, the clamp ring subassembly of grip block top fixedly connected with inner arc towards import center one side, the side is closely met with the inside front and back lateral wall face of fungi-proofing box around the grip block, grip block rear side lower extreme cup joints in the slide bar upper end through the slider, the slide bar upper end is located and has cup jointed the spring between grip block and the side fixing plate, the tangent plane of lower extreme closely meets between the grip.

Preferably, the clamping ring assembly comprises a clamping ring main body, a hollow groove, an extension portion, an elastic material passing assembly and a clamping pad, the hollow groove is formed in the clamping ring main body, the extension portion is arranged on one side, close to the inlet, of the lower end of the clamping ring main body, the extension portion is arc-shaped, the telescopic elastic material passing assembly is fixedly mounted on one side, close to the inlet, of the clamping ring main body, and the clamping pad is fixedly connected between the other ends of the elastic material passing assembly and is arc-shaped.

Preferably, elasticity is crossed the material subassembly and is including the material passing pipe, the discharge opening, extrusion rod and shutoff piece, the clamp ring main part is close to the hollow round platform form material passing pipe of clamp pad fixedly connected with, the material passing pipe bottom is located the clamp ring main part outside and is equipped with the discharge opening, sliding connection has coniform shutoff piece in the material passing pipe, the shutoff piece is close to clamp pad one side and presss from both sides fixedly connected with extrusion rod between the pad, the spring has been cup jointed to the extrusion rod upper end in the material passing pipe outside, material passing pipe cross-section diameter is close to hollow groove one side and increases step by step, shutoff piece cross-section diameter is kept away from hollow groove one.

Preferably, flexible pipe assembly is including fixed pipe, remove pipe and connection ball, the side fixing plate is equipped with the logical groove that transversely runs through, fixed pipe fixed connection is between steam manufacturing equipment and side fixing plate, it has the removal pipe to run through sliding connection in the logical inslot, it reaches fixed intraduct to remove tub one end delay, it is close to import central point and puts one end fixedly connected with connection ball to remove the pipe, connect the ball and pass through the connecting pipe intercommunication with the venthole, it is located the side fixing plate and connect and has cup jointed the spring between the ball to remove the pipe upper end, it is located side fixing plate central point and puts the subassembly that opens and shuts that is provided with the removal pipe.

Preferably, lead to the inslot center upper and lower extreme and all be equipped with the spacing groove, one side that the pipe center was kept away from to the spacing groove is established to the inclined plane, the subassembly that opens and shuts is including the separation blade, the dog, push rod and movable block, the inside equal fixedly connected with separation blade in upper end that is located both sides about spacing groove department of removal intraduct, equal sliding connection has the dog between the separation blade of removal intraduct upper end and between the separation blade of inside lower extreme, the dog is kept away from and is moved intraduct central point and put the equal fixedly connected with in one side and run through the push rod of removal outside of tubes wall, the push rod extends to spacing inslot fixedly connected with movable block, the spring has been cup jointed outside the push rod is.

A method for using stem cells for cell culture for directional differentiation, which comprises the following steps:

s1: firstly, the culture dish and the interior of the box body are disinfected and sterilized.

S2: an appropriate amount of culture medium was added to the petri dish.

S3: the culture dish is clamped into the moving groove, the stem cells are laid in the culture solution for conventional culture, and then the cover plate is clamped into the clamping groove.

S4: the prepared stem cell suspension was loaded into a pipette.

S5: insert the pipette and prevent fungus box top import, push up through the pipette and move the grip block, move flexible pipe assembly through the top when the grip block opens and make high temperature steam spout, carry out germicidal treatment to the pipette surface.

S6: pressing pipette upper end rubber ball makes the suspension drip at the auxiliary culture lid upper end, starts positive and negative motor and makes culture lid 31 upset for the suspension forms the suspension and drips and instil into in the culture dish.

S7: the bottom of the culture dish is detected by the CCD detection module, data are fed back to the single chip microcomputer controller, and then the data are transmitted to a laboratory through the single chip microcomputer controller for real-time monitoring.

S8, the positive and negative motors drive the screw rod to rotate and drive the bearing plate to move up and down, the CCD detects the cells at a plurality of positions, and the driven shaft drives the culture dish to rotate and slightly shake the culture dish, so that the stem cells are not easy to gather.

Has the advantages that:

1. drive the lead screw through positive and negative motor and rotate to make the loading board drive CCD detection module and reciprocate to the culture dish real-time detection, feed back data to the laboratory through the single chip microcomputer controller, the observation of being more convenient for, the driven shaft drives drive gear simultaneously and drives the culture dish and rotate, prevents that stem cell from gathering in the culture solution.

2. The pipette inserts the import, can push up and move the grip block swing and open, thereby push up through the grip block and move the connection ball, it moves the fixed intraductal removal of pipe network to connect the ball top, make the subassembly that opens and shuts toward fixed intraductal removal, thereby make the dog open, start-up steam manufacturing equipment, high temperature steam removes the pipe through fixed pipe entering, through removing the pipe entering connection ball, get into the venthole through the connecting pipe, then spout to the pipette, thereby disinfect to the pipette, start the aspiration pump, take out the steam after disinfecting, prevent that the steam precooling liquefaction from falling into in the fungi-proofing box.

3. When the pipette inserted the import, the top moved the clamping ring subassembly, then will push up the movable ring pad to make the extrusion stem promote the shutoff piece and push into hollow inslot, add solvent type paraffin in the hollow inslot, paraffin passes through the discharge opening and flows in the extension, thereby make the more inseparable centre gripping pipette of ring pad, secondly solvent volatilizees in the paraffin, paraffin solidifies and seals, prevents that the outside air from bringing into the bacterium and getting into fungi-proofing box.

Description of the drawings:

fig. 1 is a schematic overall front sectional view of the present invention.

Fig. 2 is a schematic view of a front cross-section mechanism of the antibacterial box of the present invention.

FIG. 3 is a schematic side view of the fungi-proofing box of the present invention.

Fig. 4 is an enlarged schematic view of a portion a in fig. 2 according to the present invention.

Fig. 5 is a schematic top view of the box of the present invention.

FIG. 6 is a schematic diagram of the top view of the driving mechanism of the culture dish of the present invention.

Fig. 7 is a schematic structural view of the clamping and air-injecting assembly of the present invention.

FIG. 8 is a schematic view of a clamp ring assembly of the present invention.

Fig. 9 is an enlarged view of the structure at B in fig. 8 according to the present invention.

In FIGS. 1-9: the device comprises a box body 1, a clamping groove 101, an air cushion 102, a cover plate 103, a culture dish 2, an annular buckle 201, an outer gear ring 202, an antibacterial box 3, an auxiliary culture cover 31, an inlet 32, a sliding groove 33, a sliding rod 34, a side fixing plate 35, a through groove 351, a limiting groove 352, a driven shaft 4, a driving gear 41, a first partition plate 5, a second partition plate 6, a bearing plate 7, a CCD detection module 71, a single-chip microcomputer controller 8, a screw rod 9, steam manufacturing equipment 10, a clamping and air injection assembly 11, a clamping plate 111, a clamping ring assembly 112, a tangent plane 113, an air outlet 114, an air inlet 12, a telescopic pipe assembly 13, a fixing pipe 131, a moving pipe 132, a connecting ball 133, an air suction pump 14, an air suction pipe 141, an opening and closing assembly 15, a baffle 151, a baffle 152, a push rod 153, a moving block 154, a connecting block 16, a moving grooves 161, a clamping ring main body 1121, a hollow, A block 194, a ring pad 20.

Detailed Description

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, rather than all embodiments, and all other embodiments obtained by a person of ordinary skill in the art without creative efforts based on the embodiments of the present invention belong to the protection scope of the present invention.

As shown in figures 1 to 9: the invention provides a technical scheme, in particular to a stem cell directional differentiation device for cell culture and a using method thereof, the device comprises a box body 1, a cover plate 103, a culture dish 2 and an antibacterial box 3, a detachable cover plate 103 is movably arranged at an opening at the top of the box body 1, the antibacterial box 3 is fixedly connected at the center of the top of the cover plate 103, the lower end of the antibacterial box 3 extends into the box body 1, connecting blocks 16 are fixedly connected at the front side and the rear side of the interior of the box body 1, the culture dish 2 capable of horizontally rotating is movably connected between the connecting blocks 16, a first partition plate 5 is fixedly connected at the lower end of the interior of the box body 1, a second partition plate 6 is fixedly connected at the upper end of the first partition plate 5 in the interior of the box body 1, a single chip microcomputer controller 8 is fixedly arranged between the first partition plate 5 and the second partition plate 6, a positive and negative motor is fixedly, positive and negative motor passes through sprocket chain synchronous linkage with lead screw 9, but lead screw 9 is located threaded connection has the loading board 7 of oscilaltion between the two 6 upper ends of baffle, 7 top fixed mounting of loading board have CCD detection module 71, CCD detection module 71 is the CCD camera, CCD detection module 71 is connected with single chip microcomputer controller 8 electricity, start positive and negative motor and drive lead screw 9 and rotate to make loading board 7 remove, can follow different positions through the CCD camera and monitor the observation.

Wherein: 6 top right side central point of baffle two puts and rotates and be connected with vertical driven

shaft

4, 4 tops of driven shaft extend to 7 upper ends outside fixedly connected with drive gear 41 of loading board, 2 outer ring gears 202 of lower extreme outside fixedly connected with of culture dish, drive gear 41 and outer ring gears 202 meshing joint, 2 top outside fixedly connected with annular buckle 201 of culture dish, shifting chute 161 runs through the groove for the arc, annular buckle 201 corresponds the inside sliding connection of shifting chute 161, the apron 103 bottom is embedded to have the universal ball of round, universal ball and the 201 top sliding connection of annular buckle, 9 pivoted in-process of lead screw drives driven shaft 4 and rotates, driven shaft 4 drives drive gear 41 and rotates, drive gear 41 drives culture dish 2 and rotates, prevent the gathering of stem cell in the culture dish 2, universal ball makes the rotation that culture dish 2 can be better.

Wherein: the top of box 1 is equipped with square form draw-in groove 101, the apron 103 corresponds the inside hollow position joint of draw-in groove 101, the air cushion 102 of the inside equal fixedly connected with intercommunication in both sides of draw-in groove 101, the air cushion 102 outside is equipped with the air inlet, through going into the draw-in groove 101 back when the apron 103 card, it inflates to aim at the air inlet through inflating equipment, thereby make air cushion 102 fill the centre gripping apron 103, make the apron 103 be difficult for breaking away from the draw-in groove, secondly, it reaches sealed effect to swell through air cushion 102.

Wherein: the antibacterial box 3 comprises an auxiliary culture cover 31, an inlet 32, a sliding groove 33, a sliding rod 34 and a side fixing plate 35, wherein the center of the top of the antibacterial box 3 is provided with the inlet 32, the bottom of the antibacterial box 3 is rotatably connected with the auxiliary culture cover 31, a positive and negative motor is fixedly connected inside the antibacterial box 3, the output shaft of the positive and negative motor is in transmission connection with the center shaft of the auxiliary culture cover 31 through a bevel gear, the left and right sides of the central position of the lower end of the inlet 32 at the rear side inside the antibacterial box 3 are respectively provided with an arc-shaped sliding groove 33, the arc-shaped sliding rod 34 is fixedly connected inside the sliding groove 33, the left and right sides of the inlet 32 are respectively and rotatably connected with a clamping air injection assembly 11 for closing the inlet 32, the left and right sides of the inlet 32 at the top inside the antibacterial box 3 are respectively and vertically and fixedly connected with the side fixing plate 35, the steam manufacturing, the telescopic pipe assembly 13 is communicated with the clamping air injection assembly 11 through a connecting pipe, a plurality of air suction holes 12 are sequentially formed in the rear side of the interior of the fungi-proofing box 3 at equal intervals, an air suction pump 14 is fixedly installed on the rear side of the fungi-proofing box 3, an air suction pipe 141 is fixedly installed between the air suction pump 14 and the air suction holes 12, the clamping plate 111 is pushed when a pipette is inserted into the inlet 32, the telescopic pipe assembly 13 is extruded through the clamping plate 111, the opening and closing assembly 15 in the telescopic pipe assembly 13 is opened, steam enters the fungi-proofing box 3 to be sterilized after the steam manufacturing equipment 10 is started, and after the air suction pump 14 is started, steam which is not condensed and liquefied is quickly led out through the air suction pipe 141 through the air suction holes 12, and.

Wherein: the clamping air injection assembly 11 comprises a clamping plate 111, a clamping ring assembly 112, a tangent plane 113 and air outlet holes 114, wherein an arc-shaped groove is formed in one side of the clamping plate 111 close to the center of the inlet 32, a plurality of air outlet holes 114 are formed in the arc-shaped groove, the tangent plane 113 is formed in one side of the bottom of the clamping plate 111 close to the center of the inlet 32, the clamping ring assembly 112 with an inner arc facing to one side of the center of the inlet 32 is fixedly connected to the top of the clamping plate 111, the front side and the rear side of the clamping plate 111 are tightly connected with the front side wall and the rear side wall inside the antibacterial box 3, the lower end of the rear side of the clamping plate 111 is sleeved at the upper end of a sliding rod 34, a spring is sleeved at the upper end of the sliding rod 34 and positioned between the clamping plate 111 and, the bacteria-proof box 3 can be isolated from being communicated with the outside in the initial state through the clamping plate 111, and bacteria can be prevented from entering.

Wherein: the clamping ring assembly 112 comprises a clamping ring main body 1121, a hollow groove 17, an extending portion 18, an elastic material passing assembly 19 and a clamping pad 20, the hollow groove 17 is formed in the clamping ring main body 112, the extending portion 18 is arranged on one side, close to the inlet 32, of the lower end of the clamping ring main body 112, the extending portion 18 is arc-shaped, the telescopic elastic material passing assembly 19 is fixedly installed on one side, close to the inlet 32, of the clamping ring main body 112, and the arc-shaped clamping pad 20 is fixedly connected between the other ends of the elastic material passing assembly 19.

Wherein: the elastic material passing assembly 19 comprises a material passing pipe 191, a discharge hole 192, an extrusion rod 193 and a blocking block 194, a hollow truncated cone-shaped material passing pipe 191 is fixedly connected to the clamping ring main body 1121 close to the clamping pad 20, the discharge hole 192 is formed in the bottom of the material passing pipe 191 and located on the outer side of the clamping ring main body 1121, a conical blocking block 194 is connected in the material passing pipe 191 in a sliding mode, the extrusion rod 193 is fixedly connected between one side, close to the clamping pad 20, of the blocking block 194 and the clamping pad 20, a spring is sleeved at the upper end of the extrusion rod 193 and located on the outer side of the material passing pipe 191, the section diameter of the material passing pipe 191 is gradually increased close to one side of the hollow groove 17, the section diameter of the blocking block 194 far away from one side of the hollow groove 17 is gradually increased, when a pipette is inserted into the inlet 32, the clamping ring assembly 112 is jacked, then the annular pad 20 is jacked, so that the extrusion, thereby make the more inseparable centre gripping pipette of ring pad 20, secondly solvent in the paraffin volatilizes, and the paraffin solidifies and seals, prevents that outside air from bringing into the bacterium and getting into fungi-proofing box 3.

Wherein: the telescopic tube assembly 13 comprises a fixed tube 131, a moving tube 132 and a connecting ball 133, wherein a through groove 351 transversely penetrating through the side fixing plate 35 is arranged on the side fixing plate 35, the fixed tube 131 is fixedly connected between the steam manufacturing equipment 10 and the side fixing plate 35, the moving tube 132 penetrates through the through groove 351 and is slidably connected with the moving tube 132, one end of the moving tube 132 extends into the fixed tube 131, one end of the moving tube 132, which is close to the central position of the inlet 32, is fixedly connected with the connecting ball 133, the connecting ball 133 is communicated with the air outlet hole 114 through a connecting pipe, a spring is sleeved between the side fixing plate 35 and the connecting ball 133 at the upper end of the moving tube 133, the opening and closing assembly 15 for opening and closing the moving tube 132 is arranged at the central position of the side fixing plate 35 on the moving tube 132, the connecting ball 133 is jacked in, the clamping plate 111 is always in close contact with the pipette by the action of the connecting ball 133 and the spring at the upper end of the slide 34.

Wherein: the upper and lower ends of the inner center of the through groove 351 are respectively provided with a limit groove 352, one side of the limit groove 352 far away from the center of the moving pipe 132 is provided with an inclined surface, the opening and closing assembly 15 comprises baffle plates 151, a stop block 152, a push rod 153 and a moving block 154, the upper ends of the moving pipe 132, which are positioned at the left and right sides of the limit groove 352, are respectively and fixedly connected with the baffle plates 151, the baffle plates 151 at the upper end of the inner part of the moving pipe 132 and the baffle plates 151 at the lower end of the inner part are respectively and slidably connected with the stop block 152, one side of the stop block 152, which is far away from the inner center of the moving pipe 132, is respectively and fixedly connected with the push rod 153 penetrating through the outer wall of the moving pipe 132, the push rod 153 extends into the limit groove 352 and is fixedly connected with the moving block 154, the spring is, the moving block 154 also moves along with the moving, under the influence of the inclined plane of the limiting groove 352 and the action of the spring at the upper end of the push rod 153, the moving block 154 drives the push rod 153 to move in the limiting groove 352, so that the stopper 152 is opened, steam enters the moving pipe 132 through the fixed pipe 131, enters the connecting ball 133 through the moving pipe 132, and finally enters the air outlet 114 through the connecting pipe to be sprayed out to sterilize the pipette.

s1: first, the culture dish 2 and the inside of the case 1 are sterilized.

S2: an appropriate amount of culture solution was added to the petri dish 2.

S3: the culture dish 2 is clamped into the moving groove 161, the stem cells are laid in the culture solution for conventional culture, and then the cover plate 103 is clamped into the clamping groove 101.

S4: the prepared stem cell suspension was loaded into a pipette.

S5: insert the import at fungi-proofing box top with the pipette, push up clamping plate 111 through the pipette, it makes high temperature steam blowout to move flexible pipe assembly 13 through the top when clamping plate 111 opens, carries out germicidal treatment to the pipette surface.

S6: pressing pipette upper end rubber ball makes the suspension drip at the supplementary culture lid 31 upper end, starts positive and negative motor and makes culture lid 31 upset for the suspension forms the suspension and drips and instil into in the culture dish 2.

S7: the bottom of the culture dish 2 is detected through the CCD detection module, data are fed back to the single-chip microcomputer controller 8, and then the data are transmitted to a laboratory through the single-chip microcomputer controller 8 for real-time monitoring.

S8, the positive and negative motors drive the screw rod to rotate and drive the bearing plate 7 to move up and down, the CCD detects the cells at a plurality of positions, and the driven shaft 4 drives the culture dish 2 to rotate and slightly shake the cells, so that the stem cells are not easy to gather.

The working principle is as follows:

firstly, a culture dish 2 and the interior of a box body 1 are disinfected and sterilized, a proper amount of culture solution is added into the culture dish 2, the culture dish 2 is clamped into a moving groove 161, stem cells are laid in the culture solution for conventional culture, then a cover plate 103 is clamped into a clamping groove 101, air is pumped by a pumping device aiming at an air inlet, so that an air cushion 102 fills up a clamping cover plate 103, the cover plate 103 is not easy to separate from the clamping groove, secondly, the air cushion 102 is bulged to achieve the sealing effect, when suspension liquid drops need to be added, a pipette is inserted into an inlet 32 by aiming at the inlet, so that the pipette is clamped by a clamping plate 111, firstly, when the pipette is inserted into the inlet 32, the clamping ring assembly 112 is jacked, then the ring gasket 20 is jacked, so that an extrusion rod 193 pushes a sealing block 194 to be pushed into the hollow groove 17, solvent type paraffin is added into the hollow groove 17, the paraffin flows into an extension part 18 through a discharge, secondly, the solvent in the paraffin is volatilized, the paraffin is solidified and sealed, bacteria brought by external air are prevented from entering the antibacterial box 3, secondly, the clamping plate 111 pushes the connecting ball 133 in the process of being pressed by the pipette, the connecting ball 133 pushes the moving pipe 132 to move towards the inside of the fixed pipe 131, when the moving pipe 132 moves towards the inside of the fixed pipe 131, the moving block 154 also moves along with the moving, under the influence of the inclined surface of the limiting groove 352 and the action of a spring at the upper end of the push rod 153, the push rod 153 is driven by the moving block 154 to move towards the limiting groove 352, so that the stoppers 152 are opened, steam enters the moving pipe 132 through the fixed pipe 131, enters the connecting ball 133 through the moving pipe 132, finally enters the air outlet hole 114 through the connecting pipe to be sprayed out to sterilize the pipette, the air pump 14 is started, the steam which is not condensed and liquefied is quickly led out through the air suction hole 12 to prevent the liquefied, then press pipette top rubber ball for the suspension drips into auxiliary culture lid 31, start positive and negative motor and drive the upset of auxiliary culture lid 31, make the suspension drip into culture dish 2, start positive and negative motor and drive lead screw 9 and rotate, thereby make loading board 7 remove, can follow different positions through the CCD camera and monitor the observation, pole 9 pivoted in-process drives driven shaft 4 and rotates, driven shaft 4 drives drive gear 41 and rotates, drive gear 41 drives culture dish 2 and rotates, prevent the aggregation of stem cells in the culture dish 2.

Claims

1. The utility model provides a directional differentiation device of stem cell for cell culture, includes box (1), apron (103), culture dish (2), fungi-proofing box (3), its characterized in that: the improved anti-bacterial incubator is characterized in that a detachable cover plate (103) is movably mounted at the top opening of the box body (1), a bacteria-proof box (3) is fixedly connected to the center of the top of the cover plate (103), the lower end of the bacteria-proof box (3) extends into the box body (1), connecting blocks (16) are fixedly connected to the front side and the rear side of the interior of the box body (1), a culture dish (2) capable of horizontally rotating is movably connected between the connecting blocks (16), a first partition plate (5) is fixedly connected to the lower end of the interior of the box body (1), a second partition plate (6) is fixedly connected to the upper end of the first partition plate (5) in the interior of the box body (1), a single-chip microcomputer controller (8) is fixedly mounted between the first partition plate (5) and the second partition plate (6), a positive and negative motor is fixedly mounted between the first partition, synchronous linkage through sprocket chain between lead screw (9), positive and negative motor passes through sprocket chain synchronous linkage with lead screw (9), but lead screw (9) are located threaded connection have bearing board (7) of oscilaltion between two (6) upper ends of baffle, bearing board (7) top fixed mounting has CCD detection module (71).

2. A stem cell differentiation directing apparatus for cell culture according to claim 1, characterized in that: baffle two (6) top right side central point puts and rotates and be connected with vertical driven shaft (4), driven shaft (4) top extends to loading board (7) upper end outside fixedly connected with drive gear (41), outer ring gear (202) of culture dish (2) lower extreme outside fixedly connected with, drive gear (41) and outer ring gear (202) meshing joint, culture dish (2) top outside fixedly connected with annular buckle (201), shifting chute (161) runs through the groove for the arc, annular buckle (201) correspond shifting chute (161) inside sliding connection, apron (103) bottom is embedded to have the universal ball of round, universal ball and annular buckle (201) top sliding connection.

3. A stem cell differentiation directing apparatus for cell culture according to claim 1, characterized in that: the box (1) top is equipped with square form draw-in groove (101), apron (103) correspond the inside cavity position joint of draw-in groove (101), the equal fixedly connected with air cushion (102) that communicate each other in draw-in groove (101) inside both sides.

4. A stem cell differentiation directing apparatus for cell culture according to claim 1, characterized in that: the anti-bacterial box (3) comprises an auxiliary culture cover (31), an inlet (32), a sliding groove (33), a sliding rod (34) and a side fixing plate (35), wherein the inlet (32) is arranged at the center of the top of the anti-bacterial box (3), the auxiliary culture cover (31) is rotatably connected to the bottom of the anti-bacterial box (3), a positive and negative motor is fixedly connected to the inside of the anti-bacterial box (3), an output shaft of the positive and negative motor is connected with the center shaft of the auxiliary culture cover (31) through a bevel gear in a transmission manner, arc-shaped sliding grooves (33) are respectively arranged on the left side and the right side of the central position of the lower end of the inlet (32) on the inner rear side of the anti-bacterial box (3), the arc-shaped sliding rod (34) is fixedly connected to the inside of the sliding groove (33), the clamping air injection assembly (11) for sealing the inlet (32) is rotatably connected to the left side and the right side of the inlet (, prevent that equal fixed mounting in fungus box (3) left and right sides has steam manufacturing equipment (10), fixed mounting has telescopic tube subassembly (13) between steam manufacturing equipment (10) and side fixing plate (35), telescopic tube subassembly (13) and centre gripping jet-propelled subassembly (11) are through the connecting pipe intercommunication, prevent that the inside rear side equidistance of fungus box (3) is equipped with a plurality of suction holes (12) in proper order, prevent that fungus box (3) rear side fixed mounting has aspiration pump (14), fixed mounting has aspiration tube (141) between aspiration pump (14) and suction hole (12).

5. A stem cell differentiation directing apparatus for cell culture according to claim 4, characterized in that: the clamping air injection assembly (11) comprises a clamping plate (111), a clamping ring assembly (112), a cutting surface (113) and an air outlet hole (114), an arc-shaped groove is arranged on one side of the clamping plate (111) close to the center of the inlet (32), a plurality of air outlet holes (114) are arranged in the arc-shaped groove, one side of the bottom of the clamping plate (111) close to the center of the inlet (32) is provided with a tangent plane (113), the top of the clamping plate (111) is fixedly connected with a clamping ring component (112) with an inner arc towards one side of the center of the inlet (32), the front side and the rear side of the clamping plate (111) are tightly connected with the front side wall and the rear side wall inside the antibacterial box (3), the lower end of the rear side of the clamping plate (111) is sleeved at the upper end of the sliding rod (34) through a sliding block, a spring is sleeved between the clamping plate (111) and the side fixing plate (35) at the upper end of the sliding rod (34), when the clamping plates (111) are in the initial position, the tangent planes (113) at the lower ends of the clamping plates (111) are tightly connected.

6. A stem cell differentiation directing apparatus for cell culture according to claim 5, characterized in that: clamping ring subassembly (112) are including clamping ring main part (1121), hollow groove (17), extension (18), elasticity material passing subassembly (19) and clamp pad (20), clamping ring main part (112) inside is equipped with hollow groove (17), clamping ring main part (112) lower extreme is close to import (32) one side and is equipped with extension (18), extension (18) are the arc form, clamping ring main part (112) are close to import (32) one side fixed mounting have telescopic elasticity material passing subassembly (19), fixedly connected with arc form clamp pad (20) between elasticity material passing subassembly (19) the other end.

7. A stem cell differentiation directing apparatus for cell culture according to claim 6, characterized in that: elasticity crosses material subassembly (19) including material passing pipe (191), discharge opening (192), extrusion stem (193) and shutoff piece (194), clamp ring main part (1121) is close to clamp pad (20) fixedly connected with hollow round platform form material passing pipe (191), material passing pipe (191) bottom is located clamp ring main part (1121) outside and is equipped with discharge opening (192), material passing pipe (191) sliding connection has coniform shutoff piece (194), shutoff piece (194) are close to clamp pad (20) one side and clamp between pad (20) fixedly connected with extrusion stem (193), the spring has been cup jointed in material passing pipe (191) outside to extrusion stem (193) upper end, material passing pipe (191) cross-section diameter is close to hollow groove (17) one side and increases step by step, shutoff piece (194) cross-section diameter is kept away from hollow groove (17) one side and is increased step by step.

8. A stem cell differentiation directing apparatus for cell culture according to claim 1, characterized in that: the telescopic pipe component (13) comprises a fixed pipe (131), a moving pipe (132) and a connecting ball (133), the side fixing plate (35) is provided with a through groove (351) which transversely penetrates through, the fixing pipe (131) is fixedly connected between the steam manufacturing equipment (10) and the side fixing plate (35), a moving pipe (132) penetrates through the through groove (351) in a sliding manner, one end of the moving pipe (132) extends into the fixed pipe (131), one end of the moving pipe (132) close to the central position of the inlet (32) is fixedly connected with a connecting ball (133), the connecting ball (133) is communicated with the air outlet (114) through a connecting pipe, a spring is sleeved between the side fixing plate (35) and the connecting ball (133) at the upper end of the moving pipe (133), the movable pipe (132) is positioned at the center of the side fixing plate (35) and is provided with an opening and closing assembly (15) for opening and closing the movable pipe (132).

9. A stem cell differentiation directing apparatus for cell culture according to claim 1, characterized in that: the upper end and the lower end of the center in the through groove (351) are respectively provided with a limiting groove (352), one side of the limiting groove (352) far away from the center of the moving pipe (132) is provided with an inclined surface, the opening and closing assembly (15) comprises baffle sheets (151), baffle blocks (152), push rods (153) and moving blocks (154), the upper ends of the left side and the right side of the limiting groove (352) in the moving pipe (132) are respectively and fixedly connected with the baffle sheets (151), the baffle blocks (152) are respectively and slidably connected between the baffle sheets (151) at the upper end in the moving pipe (132) and between the baffle sheets (151) at the lower end in the moving pipe (132), one side of the baffle blocks (152) far away from the center in the moving pipe (132) is respectively and fixedly connected with the push rods (153) penetrating through the outer wall of the moving pipe (132), the push rods (153) extend into the limiting groove (352), the side, connected with the inner side inclined plane of the limiting groove (352), of the moving block (154) is also provided with an inclined plane, and the inclined plane of the moving block (154) is rotatably connected with a roller.

10. A method for using stem cells for cell culture for directional differentiation, which comprises the following steps:

s1: firstly, the culture dish (2) and the interior of the box body (1) are disinfected and sterilized.

S2: adding a proper amount of culture solution into the culture dish (2).

S3: the culture dish (2) is clamped into the moving groove (161), the stem cells are laid in the culture solution for conventional culture, and then the cover plate (103) is clamped into the clamping groove (101).

S4: the prepared stem cell suspension was loaded into a pipette.

S5: insert the import of fungi-proofing box top with the pipette, push up through the pipette and move grip block (111), grip block (111) open the time make high temperature steam blowout through pushing up flexible pipe assembly (13), carry out germicidal treatment to the pipette surface.

S6: pressing the rubber ball at the upper end of the pipette to enable the suspension to be dripped to the upper end of the auxiliary culture cover (31), starting the positive and negative motors to enable the culture cover (31) to turn over, and enabling the suspension to form the suspension to be dripped into the culture dish (2).

S7: detect culture dish (2) bottom through CCD detection module and feed back data to single chip microcomputer controller (8), then give laboratory, real-time supervision with data transmission through single chip microcomputer controller (8).

S8, the positive and negative motors drive the screw rod to rotate and drive the bearing plate (7) to move up and down, the CCD detects the stem cells at a plurality of positions, and the driven shaft (4) drives the culture dish (2) to rotate and slightly shake the culture dish, so that the stem cells are not easy to gather.