CN114672415A

CN114672415A - Stem cell culture method and inoculation culture device thereof

Info

Publication number: CN114672415A
Application number: CN202210367633.0A
Authority: CN
Inventors: 谢建敏
Original assignee: Hubei Mingde Health Technology Co ltd
Current assignee: Hubei Mingde Health Technology Co ltd
Priority date: 2022-04-08
Filing date: 2022-04-08
Publication date: 2022-06-28
Anticipated expiration: 2042-04-08
Also published as: CN114672415B

Abstract

The invention discloses a stem cell culture method and an inoculation culture device thereof, relating to the technical field of stem cell inoculation culture; in order to improve the reliability and convenience of dilution operation; the device comprises a base, wherein a diluting mechanism is arranged on the base and comprises a supporting component and a plurality of diluting components, and the diluting components are arranged on the supporting component; the dilution assembly comprises: the outer wall of one side of the first dilution cylinder is provided with scale marks, and the top of the first dilution cylinder is detachably provided with an ejector rod; the method comprises the following steps: sampling a bacterial liquid sample, wherein the using amount is equal to 1 time of the volume of the quantitative conveying cavity; and (3) injecting diluent into the first diluting cylinder and the second diluting cylinder, wherein the injection amount of the diluent is 9 times of the volume of the quantitative conveying cavity. The method is simple to operate, multiple measurement is not needed in the dilution process, the accuracy of dilution multiple is guaranteed, external air contact is avoided, other bacterial colonies are effectively prevented from being mixed, and the reliability is improved.

Description

Stem cell culture method and inoculation culture device thereof

Technical Field

The invention relates to the technical field of stem cell inoculation culture, in particular to a stem cell culture method and an inoculation culture device thereof.

Background

Stem cells have the unique ability to self-renew or differentiate into multiple cell types in response to appropriate signals. These properties make stem cells have a strong ability to repair, replace and regenerate tissues, and human stem cells are of great interest in medical research; in the stem cell culture process, bacterial liquid needs to be diluted and then placed in a culture dish for culture; during the dilution operation, need take repeatedly and place in new diluent to reach appointed dilution concentration, though can satisfy certain user demand, the complex operation process is inconvenient, and sneaks into other bacterial colonies in the air easily, consequently, still remains to be improved.

Through search, the chinese patent application No. CN202020509125.8 discloses an inoculating loop with a diluted bacteria liquid groove, the inoculating loop is made of integrally formed glass, the inoculating loop is provided with a handle, a ring and a loop rod arranged between the handle and the ring, one end of the loop rod is connected to one side outer diameter of the ring, a bacteria liquid diluting shallow groove which is circumferentially distributed on the outer diameter surface of the ring is arranged at the other side of the ring and is positioned at the thickness of the ring 1/2, the bacteria liquid diluting shallow groove crosses the half peripheral surface of the other side of the ring, the bacteria liquid diluting shallow groove is provided with two first side walls which are parallel to the end surface of the ring and two second side walls which are vertical to the end surface of the ring, the first side walls and the second side walls are connected with each other and the first side walls and the outer diameter surface of the ring through smooth chamfers, and the second side wall is vertical to the outer diameter of the ring and the first side wall is vertical to the bottom surface of the bacteria liquid diluting shallow groove. The inoculating loop in the above patent has the following disadvantages: although certain use requirements can be met, the dilution operation is inconvenient, the dilution rate cannot be controlled accurately easily, and other bacterial colonies in the air are mixed easily, so that the improvement is needed.

Disclosure of Invention

The invention aims to solve the defects in the prior art and provides a stem cell culture method and an inoculation culture device thereof.

In order to achieve the purpose, the invention adopts the following technical scheme:

a stem cell inoculation culture device comprises a base, wherein a dilution mechanism is arranged on the base and comprises a support component and a plurality of dilution components, and the dilution components are arranged on the support component; the dilution assembly comprises:

the outer wall of one side of the first dilution cylinder is provided with scale marks, and the top of the first dilution cylinder is detachably provided with an ejector rod;

the first quantitative conveying chambers are respectively arranged at the bottom of the first dilution cylinder, and are alternately distributed with the first dilution cylinder; the upper and lower outer walls of the first quantitative conveying chamber are provided with through holes, and the interior of the first quantitative conveying chamber is communicated with the interiors of the two connected first dilution barrels through the upper and lower through holes;

the first conduction control part comprises a rotating disc and a separating block, the rotating disc is fixed at the central position of one side of the separating block, and the rotating disc is rotatably arranged at one side of the first quantitative conveying chamber; a sealing rubber gasket is fixed on the outer wall of the separation block, and the separation block rotates on the inner wall of the first quantitative conveying chamber in a sealing mode through the sealing rubber gasket; the separating block is in a butterfly structure matched with the first quantitative conveying chamber, and two symmetrical quantitative conveying cavities are formed between the separating block and the inner wall of the first quantitative conveying chamber.

Preferably: the dilution assembly further comprises:

the connecting chamber is arranged below the first quantitative conveying chamber at the bottommost part and is communicated with the first quantitative conveying chamber through the through opening;

the structure of the second diluting cylinder is the same as that of the first diluting cylinder, and the second diluting cylinder at the lowest part is communicated with the connecting chamber through a connecting pipe;

the structure of the second quantitative conveying chamber is the same as that of the first quantitative conveying chamber, the second quantitative conveying chamber is arranged above the second dilution cylinder, and the second quantitative conveying chamber and the second dilution cylinder are alternately distributed;

and the second conduction control part is arranged at the second quantitative conveying chamber and has the same structure as the first conduction control part.

Further: the support assembly includes:

the first rotating seat is rotatably arranged on the base;

the support frame is fixed at the center of the top of the first rotating seat;

the cross rods are fixed on the outer walls of the two sides of the top end of the support frame;

the fixing frame is rotatably arranged on the outer wall of the cross rod, and a first diluting cylinder and a second diluting cylinder are respectively fixed on the inner walls of the two ends of the fixing frame;

the fixing piece is arranged on the fixing frame and used for fixing the fixing frame on the cross rod and limiting the angle of the fixing frame.

Further preferably: the fixing member includes:

the fixing pin, the cross bar and the fixing frame are provided with pin holes, and the fixing pin is detachably inserted in the pin holes;

two ends of the spring are respectively fixed at one end of the fixing pin and the outer wall of one side of the fixing frame;

and the pull ring is fixed at the top end of the fixing pin.

As a preferable aspect of the present invention: the outer wall of one side of first dilution section of thick bamboo and second dilution section of thick bamboo all is provided with leads to the liquid pipe, it is connected with the valve body to lead to the liquid intraductal rotation, and valve body one end is fixed with control knob.

Further preferred as the invention: a glue film is fixed on the inner wall of one end of the liquid through pipe, a micro-seam is formed on the glue film, and the micro-seam is kept in a closed state under the condition that the glue film is not deformed; lead to liquid pipe cooperation and use and have L type intubate, L type intubate top is provided with the liquid feeding fill, and L type intubate detachable pegs graft in leading to the liquid intraductal.

As a still further scheme of the invention: install on the base and cultivate the mechanism, it includes to cultivate the mechanism:

the second rotating seat is rotatably arranged at the top of the base;

the culture chamber is fixed on the outer wall of the top of the second rotating seat;

and the culture dish is placed on the inner side of the culture chamber.

On the basis of the scheme: and a uniformly distributed check curtain is fixed on the outer wall of one side of the culture chamber.

On the basis of the foregoing scheme, it is preferable that: and a temperature control assembly is fixed on the inner wall of one side of the culture chamber.

A stem cell culture method comprises the following steps:

s1: sampling a bacterial liquid sample, wherein the using amount is equal to 1 time of the volume of the quantitative conveying cavity;

s2: injecting diluent into the first diluting cylinder and the second diluting cylinder, wherein the injection amount of the diluent is 9 times of the volume of the quantitative conveying cavity;

s3: injecting sample bacteria liquid into the uppermost first dilution cylinder;

s4: after fully mixing, screwing a rotating disc below the first dilution cylinder, and discharging the diluted bacteria liquid in the first dilution cylinder into the next first dilution cylinder through a quantitative conveying cavity, wherein the discharge amount is the volume of the quantitative conveying cavity;

s5: repeating the steps until the dilution reaches the designated multiple;

s6: taking out the diluted bacteria liquid from the liquid through pipe;

s7: after the coater is sterilized, dipping the taken bacterial liquid;

s8: the bacterial liquid is coated on a culture medium with a culture dish by using a coater for culture.

The beneficial effects of the invention are as follows:

1. the invention can inject the diluent into each first diluting cylinder when diluting, because the volume of the quantitative conveying cavity is a fixed value, the diluent injected into the first diluting cylinder is controlled to be a fixed multiple of the volume of the quantitative conveying cavity, such as 9 times, the rotating disc is rotated to drive the separating block to rotate, the quantitative conveying cavity is firstly communicated with the first diluting cylinder above, after the liquid above enters and fills the quantitative conveying cavity, the rotating disc is rotated to communicate the quantitative conveying cavity with the first diluting cylinder below, the liquid in the quantitative conveying cavity is discharged into the first diluting cylinder below and is mixed with the diluent with the corresponding multiple, thereby achieving the purpose of diluting, the steps are repeated until the dilution reaches the designated multiple, the operation is simple, and the measurement is not needed for many times in the diluting process, the accuracy of dilution multiple is ensured, and outside air contact is avoided, other bacterial colonies are effectively prevented from being mixed, and the reliability is improved.

2. By arranging the second dilution cylinder, the second quantitative conveying chamber and other structures, the dilutable multiple can be improved, the condition that the overall structure is overlong is avoided, the space is saved, and the practicability is improved; through setting up mount, horizontal pole isotructure, the rotation that can be convenient dilutes the subassembly to do benefit to and reverse the upper and lower both ends of a first dilution section of thick bamboo and a second dilution section of thick bamboo, thereby be convenient for control its inside liquid and flow according to the assigned direction, promoted the practicality.

3. Through setting up fixed pin, spring isotructure, can insert the fixed pin in the pinhole according to the demand to carry on spacingly to the angle that dilutes the subassembly, when the subassembly is diluted in needs upset, only need with the fixed pin extract from the pinhole, with the mount rotatory 180 back again with the fixed pin insert in the pinhole can, promoted the practicality.

4. By arranging the control knob, the liquid through pipe and other structures, liquid can be conveniently added and taken from the liquid through pipe, and the practicability is improved; through setting up liquid feeding fill, L type intubate and glued membrane isotructure, can be according to the demand, with L type intubate insert the micro-crack of glued membrane in, with liquid feeding fill with lead to liquid pipe and switch on mutually, and then can be convenient from the liquid feeding fill liquid feeding, promoted the practicality.

Drawings

FIG. 1 is a schematic structural diagram of a stem cell inoculation and culture device according to the present invention;

FIG. 2 is a schematic structural diagram of a cross section of a culture chamber of a stem cell inoculation culture device provided by the invention;

FIG. 3 is a schematic structural diagram of a dilution mechanism of a stem cell inoculation and culture device according to the present invention;

FIG. 4 is a schematic structural diagram of a dilution assembly of a stem cell inoculation and culture device provided by the invention;

FIG. 5 is a schematic cross-sectional structural diagram of a first dilution cylinder and a first quantitative transfer chamber of a stem cell inoculation and culture device according to the present invention;

FIG. 6 is a schematic structural diagram of a stem cell inoculation and culture device according to the present invention, which is sectioned after the L-shaped cannula and the filling tube are detached;

fig. 7 is a schematic structural diagram of a stem cell inoculation and culture device fixing frame according to the present invention.

In the figure: the device comprises a base 1, a first rotating seat 2, a support frame 3, a first diluting barrel 4, a first quantitative conveying chamber 5, a liquid feeding hopper 6, a culture chamber 7, a blocking curtain 8, a second rotating seat 9, a rotating disc 10, a temperature control assembly 11, a culture dish 12, a cross rod 13, a top rod 14, a through hole 15, a second quantitative conveying chamber 16, a connecting pipe 17, a connecting chamber 18, a control knob 19, scale marks 20, a second diluting barrel 21, a fixing frame 22, a sealing rubber pad 23, a separation block 24, a quantitative conveying cavity 25, a cannula 26L, a liquid through pipe 27, a 28 micro-seam, a valve body 29, a rubber film 30, a pin hole 31, a fixing pin 32, a spring 33 and a pull ring 34.

Detailed Description

The technical solution of the present patent will be described in further detail with reference to the following embodiments.

Reference will now be made in detail to embodiments of the present patent, examples of which are illustrated in the accompanying drawings, wherein like or similar reference numerals refer to the same or similar elements or elements having the same or similar function throughout. The embodiments described below with reference to the drawings are exemplary only for the purpose of explaining the present patent and are not to be construed as limiting the present patent.

In the description of this patent, it is to be understood that the terms "center," "upper," "lower," "front," "rear," "left," "right," "vertical," "horizontal," "top," "bottom," "inner," "outer," and the like are used in the orientations and positional relationships indicated in the figures, which are based on the orientations and positional relationships shown in the figures, and are used for convenience in describing the patent and for simplicity in description, but do not indicate or imply that the device or element being referred to must have a particular orientation, be constructed in a particular orientation, and be operated, and therefore should not be construed as limiting the patent.

In the description of this patent, it is noted that unless otherwise specifically stated or limited, the terms "mounted," "connected," and "disposed" are to be construed broadly and can include, for example, fixedly connected, disposed, detachably connected, disposed, or integrally connected and disposed. The specific meaning of the above terms in this patent may be understood by those of ordinary skill in the art as appropriate.

Example 1:

a stem cell inoculation culture device is shown in figures 1-7 and comprises a base 1, wherein a dilution mechanism is arranged on the base 1, the dilution mechanism comprises a support component and a plurality of dilution components, and the dilution components are arranged on the support component; the dilution assembly comprises:

the device comprises a first dilution cylinder 4, wherein scale marks 20 are arranged on the outer wall of one side of the first dilution cylinder 4, and a top rod 14 is detachably arranged on the first dilution cylinder 4 at the top;

the first quantitative conveying chambers 5 are respectively and integrally arranged at the bottom of the first dilution cylinder 4, and the first quantitative conveying chambers 5 and the first dilution cylinder 4 are alternately distributed; the upper outer wall and the lower outer wall of the first quantitative conveying chamber 5 are provided with through holes 15, and the inside of the first quantitative conveying chamber 5 is communicated with the insides of the two connected first dilution cylinders 4 through the upper through hole 15 and the lower through hole 15;

the first conduction control part comprises a rotating disc 10 and a separating block 24, the rotating disc 10 is fixed at the central position of one side of the separating block 24, and the rotating disc 10 is rotatably arranged at one side of the first quantitative conveying chamber 5; the outer wall of the separation block 24 is bonded with a sealing rubber gasket 23, and the separation block 24 rotates on the inner wall of the first quantitative conveying chamber 5 in a sealing way through the sealing rubber gasket 23; the separating block 24 is in a butterfly structure matched with the first quantitative conveying chamber 5, and two symmetrical quantitative conveying cavities 25 are formed between the separating block 24 and the inner wall of the first quantitative conveying chamber 5; the rotating disc 10 can drive the separating block 24 to rotate, so that the position of the quantitative conveying cavity 25 is changed, and the quantitative conveying cavity can be communicated with the first diluting cylinder 4 at the top or the first diluting cylinder 4 at the bottom;

by arranging the first diluting cylinders 4, the first quantitative conveying chamber 5, the separating block 24 and other structures, when dilution treatment is carried out, diluent can be injected into each first diluting cylinder 4, because the volume of the quantitative conveying cavity 25 is a fixed value, the diluent injected into the first diluting cylinder 4 is controlled to be a fixed multiple of the volume of the quantitative conveying cavity 25, for example, when 9 times, the rotating disc 10 is rotated to drive the separating block 24 to rotate, the quantitative conveying cavity 25 is firstly communicated with the first diluting cylinder 4 above, after the liquid above enters and fills the quantitative conveying cavity 25, the rotating disc 10 is rotated to ensure that the quantitative conveying cavity 25 is communicated with the first diluting cylinder 4 below, the liquid in the quantitative conveying cavity 25 is discharged into the first diluting cylinder 4 below and is mixed with the diluent with the corresponding multiple, so that the dilution purpose is achieved, the steps are repeated until the diluent is diluted to a designated multiple, and the mode is simple to operate, need not many times to measure at the dilution in-process, ensured the degree of accuracy of dilution multiple, and do not give the outside air contact, effectively prevented that other bacterial colonies from sneaking into, promoted the reliability.

For better spatial layout; as shown in fig. 3 and 4, the dilution unit further includes:

a connection chamber 18, the connection chamber 18 being integrally provided below the first constant-volume delivery chamber 5 at the lowermost portion, the connection chamber 18 being in communication with the first constant-volume delivery chamber 5 through the port 15;

a second dilution cylinder 21, the structure of the second dilution cylinder 21 is the same as that of the first dilution cylinder 4, and the second dilution cylinder 21 at the lowest part is communicated with the connecting chamber 18 through a connecting pipe 17;

the structure of the second quantitative conveying chamber 16 is the same as that of the first quantitative conveying chamber 5, the second quantitative conveying chamber 16 is integrally arranged above the second dilution cylinder 21, and the second quantitative conveying chamber 16 and the second dilution cylinder 21 are alternately distributed;

a second conduction control part which is arranged at the second quantitative conveying chamber 16 and has the same structure with the first conduction control part;

through setting up the second and diluting a section of thick bamboo 21, the quantitative transport chamber 16 isotructures of second, can promote the multiple that can dilute, and avoided the condition of overall structure overlength, practiced thrift the space, promoted the practicality.

For convenience of operation; as shown in fig. 3 and 4, the support assembly includes:

the first rotating seat 2, the first rotating seat 2 can be rotatably arranged on the base 1;

the support frame 3 is fixed at the center of the top of the first rotating seat 2 through screws;

the cross rods 13 are fixed on the outer walls of the two sides of the top end of the support frame 3;

the fixing frame 22 is rotatably arranged on the outer wall of the cross rod 13, and the first diluting cylinder 4 and the second diluting cylinder 21 are respectively fixed on the inner walls of two ends of the fixing frame 22;

the fixing piece is arranged on the fixing frame 22 and used for fixing the fixing frame 22 on the cross rod 13 and limiting the angle of the fixing frame 22;

through setting up structures such as mount 22, horizontal pole 13, the rotation that can be convenient dilutes the subassembly to do benefit to and reverse the upper and lower both ends of first dilution section of thick bamboo 4 and second dilution section of thick bamboo 21, thereby be convenient for control its inside liquid and flow according to the assigned direction, promoted the practicality.

To facilitate securing the mount 22; as shown in fig. 7, the fixing member includes:

the fixing pin 32, the cross bar 13 and the fixing frame 22 are provided with pin holes 31, and the fixing pin 32 is detachably inserted in the pin holes 31;

a spring 33, wherein two ends of the spring 33 are respectively fixed on one end of the fixing pin 32 and one side outer wall of the fixing frame 22;

a pull ring 34, wherein the pull ring 34 is fixed at the top end of the fixed pin 32;

through setting up fixed pin 32, spring 33 isotructure, can be as required, insert the pinhole 31 with fixed pin 32 in to carry on spacingly to the angle that dilutes the subassembly, when the subassembly is diluted in the needs upset, only need with fixed pin 32 follow pinhole 31 in extract, with the rotatory 180 back of mount 22 again with fixed pin 32 insert in pinhole 31 can, promoted the practicality.

To facilitate the liquid adding and taking; as shown in fig. 4-6, a liquid flowing pipe 27 is arranged on the outer wall of one side of each of the first diluting cylinder 4 and the second diluting cylinder 21, a valve body 29 is rotatably connected to the liquid flowing pipe 27, and a control knob 19 is fixed at one end of the valve body 29; through setting up structures such as control knob 19, liquid pipe 27, can be convenient get liquid from liquid pipe 27 liquid feeding, promoted the practicality.

Liquid is taken out for more convenient liquid adding; as shown in fig. 5 and 6, a glue film 30 is adhered to an inner wall of one end of the liquid through pipe 27, a micro-slit 28 is formed on the glue film 30, and the micro-slit 28 is kept in a sealed state under the condition that the glue film 30 is not deformed; the liquid passing pipe 27 is matched with an L-shaped insertion pipe 26, the liquid adding hopper 6 is integrally arranged at the top end of the L-shaped insertion pipe 26, and the L-shaped insertion pipe 26 is detachably inserted into the liquid passing pipe 27; through setting up liquid feeding fill 6, L type intubate 26 and glued membrane 30 isotructure, can insert L type intubate 26 in the micro-crack 28 of glued membrane 30 according to the demand, with liquid feeding fill 6 with lead to liquid pipe 27 and switch on mutually, and then can be convenient from the liquid feeding fill 6 liquid feeding, promoted the practicality.

Example 2:

a stem cell inoculation culture device is shown in figure 1 and figure 2, and is convenient for culture; the present embodiment is modified from embodiment 1 in the following way: install on the base 1 and cultivate the mechanism, it includes to cultivate the mechanism:

the second rotating seat 9, the second rotating seat 9 can be rotatably installed on the top of the base 1;

the culture chamber 7 is fixed on the outer wall of the top of the second rotating seat 9;

a culture dish 12, the culture dish 12 being placed inside the culture chamber 7.

In order to improve the practicability; as shown in fig. 1, the outer wall of one side of the culture chamber 7 is bonded with uniformly distributed baffles 8, and the inner wall of one side of the culture chamber 7 is fixed with a temperature control assembly 11 through screws; through setting up the check curtain 8, can place the pollution, for cultivateing providing the guarantee, set up accuse temperature subassembly 11 and can be convenient for cultivate the cultivation temperature in the room 7 to the accuse, promoted the practicality.

Example 3:

a stem cell culture method comprises the following steps:

s1: sampling bacterial liquid, wherein the using amount is equal to 1 time of the volume of the quantitative conveying cavity 25;

s2: injecting a diluent into the first dilution cylinder 4 and the second dilution cylinder 21, wherein the injection amount of the diluent is 9 times of the volume of the quantitative delivery cavity 25;

s3: injecting sample bacteria liquid into the uppermost first dilution cylinder 4;

s4: after being fully mixed, the diluted bacteria liquid in the first dilution cylinder 4 is discharged into the next first dilution cylinder 4 through the quantitative conveying cavity 25 by screwing the lower rotating disc 10, and the discharge amount is the volume of the quantitative conveying cavity 25;

s5: repeating the steps until the dilution reaches the designated multiple;

s6: taking out the diluted bacteria liquid from the liquid through pipe 27;

s7: after the coater is sterilized, dipping the taken bacterial liquid;

s8: the culture medium with the petri dish 12 is coated with the bacterial liquid by using a coater to culture.

The above description is only for the preferred embodiment of the present invention, but the scope of the present invention is not limited thereto, and any person skilled in the art should be considered to be within the technical scope of the present invention, and the technical solutions and the inventive concepts thereof according to the present invention should be equivalent or changed within the scope of the present invention.

Claims

1. A stem cell inoculation culture device comprises a base (1), and is characterized in that a dilution mechanism is mounted on the base (1), the dilution mechanism comprises a support component and a plurality of dilution components, and the dilution components are mounted on the support component; the dilution assembly comprises:

the device comprises a first dilution barrel (4), scale marks (20) are arranged on the outer wall of one side of the first dilution barrel (4), and a top rod (14) is detachably arranged on the first dilution barrel (4) at the top;

the first quantitative conveying chambers (5) are respectively arranged at the bottoms of the first dilution cylinders (4), and the first quantitative conveying chambers (5) and the first dilution cylinders (4) are alternately distributed; the upper outer wall and the lower outer wall of the first quantitative conveying chamber (5) are provided with through holes (15), and the interior of the first quantitative conveying chamber (5) is communicated with the interiors of two connected first diluting barrels (4) through the upper through hole and the lower through hole (15);

the first conduction control part comprises a rotating disc (10) and a separating block (24), the rotating disc (10) is fixed at the central position of one side of the separating block (24), and the rotating disc (10) is rotatably arranged at one side of the first quantitative conveying chamber (5); a sealing rubber gasket (23) is fixed on the outer wall of the separation block (24), and the separation block (24) rotates on the inner wall of the first quantitative conveying chamber (5) in a sealing mode through the sealing rubber gasket (23); the separating block (24) is in a butterfly structure matched with the first quantitative conveying chamber (5), and two symmetrical quantitative conveying cavities (25) are formed between the separating block (24) and the inner wall of the first quantitative conveying chamber (5).

2. A stem cell seeding and culturing device according to claim 1, wherein the dilution assembly further comprises:

a connecting chamber (18), wherein the connecting chamber (18) is arranged below the first quantitative conveying chamber (5) at the bottommost part, and the connecting chamber (18) is communicated with the first quantitative conveying chamber (5) through a through opening (15);

the structure of the second dilution cylinder (21) is the same as that of the first dilution cylinder (4), and the second dilution cylinder (21) at the lowest part is communicated with the connecting chamber (18) through a connecting pipe (17);

the structure of the second quantitative conveying chamber (16) is the same as that of the first quantitative conveying chamber (5), the second quantitative conveying chamber (16) is arranged above the second dilution cylinder (21), and the second quantitative conveying chamber (16) and the second dilution cylinder (21) are alternately distributed;

and a second conduction control part which is arranged at the second quantitative conveying chamber (16) and has the same structure with the first conduction control part.

3. A stem cell seeding and culturing device according to claim 2, wherein the support assembly comprises:

the first rotating seat (2), the first rotating seat (2) is rotatably arranged on the base (1);

the support frame (3), the support frame (3) is fixed in the center position of the top of the first rotating seat (2);

the cross rods (13), the cross rods (13) are fixed on the outer walls of the two sides of the top end of the support frame (3);

the fixing frame (22), the fixing frame (22) is rotatably arranged on the outer wall of the cross rod (13), and a first diluting cylinder (4) and a second diluting cylinder (21) are respectively fixed on the inner walls of the two ends of the fixing frame (22);

the fixing piece is arranged on the fixing frame (22) and used for fixing the fixing frame (22) on the cross rod (13) and limiting the angle of the fixing frame (22).

4. The stem cell seeding and culturing device according to claim 3, wherein the fixing member comprises:

the fixing pin (32), the cross rod (13) and the fixing frame (22) are provided with pin holes (31), and the fixing pin (32) is detachably inserted in the pin holes (31);

the two ends of the spring (33) are respectively fixed at one end of the fixing pin (32) and the outer wall of one side of the fixing frame (22);

and the pull ring (34), the pull ring (34) is fixed on the top end of the fixed pin (32).

5. The stem cell inoculation and culture device according to claim 4, wherein a liquid passing pipe (27) is arranged on one side of the outer wall of each of the first dilution cylinder (4) and the second dilution cylinder (21), a valve body (29) is rotatably connected to the liquid passing pipe (27), and a control knob (19) is fixed at one end of the valve body (29).

6. The stem cell inoculation culture device according to claim 5, wherein a rubber film (30) is fixed on the inner wall of one end of the liquid through pipe (27), a micro-slit (28) is formed in the rubber film (30), and the micro-slit (28) is kept in a sealed state under the condition that the rubber film (30) is not deformed; the liquid passing pipe (27) is matched with an L-shaped insertion pipe (26), the top end of the L-shaped insertion pipe (26) is provided with a liquid adding hopper (6), and the L-shaped insertion pipe (26) is detachably inserted into the liquid passing pipe (27).

7. A stem cell inoculation and culture device according to claim 6, characterized in that the base (1) is provided with culture means, the culture means comprising:

the second rotating seat (9), the second rotating seat (9) is rotatably arranged at the top of the base (1);

the culture chamber (7), the culture chamber (7) is fixed on the outer wall of the top of the second rotating seat (9);

a culture dish (12), wherein the culture dish (12) is arranged inside the culture chamber (7).

8. The stem cell inoculation and culture device according to claim 7, wherein the outer wall of one side of the culture chamber (7) is fixed with a uniformly distributed curtain (8).

9. The stem cell inoculation and culture device according to claim 8, wherein a temperature control assembly (11) is fixed on the inner wall of one side of the culture chamber (7).

10. A stem cell culture method is characterized by comprising the following steps:

s1: sampling bacterial liquid, wherein the taking amount is equal to 1 time of the volume of the quantitative conveying cavity (25);

s2: injecting a diluent into the first dilution cylinder (4) and the second dilution cylinder (21), wherein the injection amount of the diluent is 9 times of the volume of the quantitative conveying cavity (25);

s3: injecting sample bacteria liquid into the uppermost first dilution cylinder (4);

s4: after being fully mixed, the diluted bacterium liquid in the first dilution cylinder (4) is discharged into the next first dilution cylinder (4) through a quantitative conveying cavity (25), and the discharge amount is the volume of the quantitative conveying cavity (25);

s5: repeating the steps until the dilution reaches the designated multiple;

s6: taking out the diluted bacteria liquid from the liquid through pipe (27);

s7: after the coater is sterilized, dipping the taken bacterial liquid;

s8: the bacterial liquid is spread on a culture medium with a culture dish (12) by using a spreader to culture.