CN112300935A

CN112300935A - A stem cell separation and culture device

Info

Publication number: CN112300935A
Application number: CN202011220633.5A
Authority: CN
Inventors: 湛振键; 齐国光; 刘世豪
Original assignee: Guangdong Kangdun Bioengineering Technology Co Ltd
Current assignee: Guangdong Kangdun Innovation Industry Group Co ltd
Priority date: 2020-11-05
Filing date: 2020-11-05
Publication date: 2021-02-02

Abstract

The invention provides a stem cell separation and culture device, comprising a separation cylinder and an incubator, the separation cylinder includes a separation cavity and a liquid storage cavity, a filter screen is arranged in the separation cavity, the separation cavity is separated from the liquid storage cavity by a separation piston, and the separation cavity passes through The second output port is connected with a catheter, the output end of the catheter is connected with the incubator, the separation cylinder is provided with a seal, a first input port and a first output port, and the separation cavity is provided with a single-phase baffle that opens inwards , the single-phase baffle is respectively matched with the first input port and the first output port, the liquid storage chamber is connected with the first input port pipe, the incubator is rotatably connected with a fixed plate, the fixed plate is fixed with a number of culture dishes, and the catheter will Stem cell-containing medium was introduced into several petri dishes. The present invention realizes the separation of stem cells and cleaning liquid and the cultivation of stem cells through the cooperation of the separation chamber and the incubator. The separation and cultivation of stem cells are continuous, and the stem cells do not need to be manually transferred, which effectively avoids the stem cells from being polluted during the transfer process. risks of.

Description

Stem cell isolated culture device

Technical Field

The invention relates to the technical field of stem cell culture, in particular to a stem cell separation culture device.

Background

Stem cells are cells which can self-renew and proliferate and differentiate without limitation under certain conditions, can produce daughter cells with the same phenotype and genotype as themselves, can produce specialized cells which form body tissues and organs, and can also differentiate into progenitor cells.

In the process of obtaining the stem cells, the stem cells need to be cleaned by using a cleaning solution, the stem cells need to be separated from the cleaning solution after the stem cells are cleaned, the separated stem cells are manually transferred to a culture dish for culture, and the stem cells are polluted and the culture of the stem cells is influenced in the transfer process of the stem cells.

Disclosure of Invention

Therefore, the stem cell separation and culture device provided by the invention realizes the separation of stem cells and cleaning fluid and the culture of the stem cells by the matching of the separation cavity and the culture box, the separation and the culture of the stem cells are coherent, the stem cells do not need to be transferred manually, and the risk of pollution of the stem cells in the transfer process is effectively avoided.

The technical scheme of the invention is realized as follows:

a stem cell separation and culture device comprises a separation cylinder and an incubator, wherein the separation cylinder comprises a separation cavity and a liquid storage cavity, a filter screen is arranged in the separation cavity, the separation cavity is separated from the liquid storage cavity through a separation piston, the separation cavity is communicated with a liquid guide pipe through a second output port, the output end of the liquid guide pipe is communicated with the incubator, a sealing piece used for closing or opening the second output port is arranged above the separation cylinder, a first input port is arranged at the side edge of the separation cavity, a first output port is arranged at the bottom of the separation cavity, the separation cavity is provided with a single-phase baffle plate which is opened inwards, the single-phase baffle plate is respectively matched with the first input port and the first output port, the output end of the liquid storage cavity is connected with the first input port through a pipe, a fixing plate is rotationally connected in the incubator, and a plurality of culture dishes are fixed on the fixing plate, the catheter introduces culture medium containing stem cells into a plurality of culture dishes.

Preferably, the sealing element comprises an electromagnet, an electromagnetic switch, a first pressure spring and a sealing ring, the electromagnetic switch is fixed above the separating cylinder, the first pressure spring is arranged between the electromagnetic switch and the electromagnet, the sealing ring is connected with the electromagnet, and the sealing ring is used for closing or opening the second output port.

Preferably, the liquid guide pipe comprises an annular pipe and a connecting pipe, the annular pipe is arranged on the periphery of the outer part of the separation cavity, the annular pipe is communicated with the second output port, the connecting pipe is communicated with the output end of the annular pipe, and the connecting pipe is communicated with the incubator.

Preferably, the bottom of the incubator is provided with a first motor, and an output shaft of the first motor is connected with the fixing plate.

Preferably, the fixed plate is a circular fixed plate, the culture dishes are annularly fixed on the fixed plate, the tail end of the connecting pipe is communicated with a plurality of output pipes for guiding a culture medium containing stem cells into the culture dishes, and the output pipes are fixed at the top of the incubator.

Preferably, the periphery of the incubator is provided with an observation window and a taking-out port.

Preferably, the catheter is communicated with the incubator through a temporary storage box, a stirring device used for uniformly distributing the stem cells in the culture medium is arranged in the temporary storage box, the stirring device comprises a second motor and a plurality of stirring rods, an output shaft of the second motor is connected with a stirring shaft, and the stirring rods are connected with the stirring shaft.

Preferably, the first magnet is arranged on the periphery of the inner part of the separation piston, and a driving device for driving the separation piston to move up and down is arranged on the outer side of the separation cylinder.

Preferably, drive arrangement includes second magnet, lead screw, solid fixed ring and third motor, the second magnet is located gu fixed ring's inside, gu fixed ring sliding connection in the outside of cylinder, the output shaft of third motor with the lead screw is connected, the lead screw with gu fixed ring threaded connection, the second magnet with first magnet mutually supports.

Compared with the prior art, the invention has the beneficial effects that:

according to the stem cell separation and culture device provided by the invention, the separation of the stem cells and the cleaning solution and the culture of the stem cells can be realized through the matching of the separation cavity and the culture box, the separation and the culture of the stem cells are coherent, the stem cells do not need to be transferred manually, and the risk that the stem cells are polluted in the transfer process is effectively avoided; the filter screen is reversely flushed upwards by the culture medium, so that the stem cells adhered to the filter screen during separation can be attached to the culture medium, the remaining of the stem cells in the separation cavity is reduced, and the waste of the stem cells is caused; when the stem cell liquid is separated, the single-phase baffle is located a first input port, the first input port is in a closed state, the first output port at the bottom of the separation chamber is in an open state, the stem cell cleaning liquid can conveniently flow into the bottom of the separation chamber from the first output port, the separation of the stem cells and the cleaning liquid is realized, when the culture medium is pressed into the first input port, the single-phase baffle rotates in the separation chamber to open the first input port, meanwhile, the single-phase baffle is located the first output port, the first output port is closed, the mixture of the culture medium and the cleaning liquid is avoided, and the culture medium backflushing filter.

Drawings

FIG. 1 is a schematic cross-sectional view of a stem cell isolation and culture device according to the present invention;

FIG. 2 is an enlarged schematic view of a portion A of FIG. 1;

FIG. 3 is an enlarged schematic view of a portion B of FIG. 1;

FIG. 4 is a schematic front view of the incubator of the present invention;

FIG. 5 is a schematic top view of a fixing plate according to the present invention;

in the figure, 1 a separating cylinder, 2 an incubator, 3 a separating chamber, 4 a liquid storage chamber, 5 a filter screen, 6 a separating piston, 7 a first input port, 8 a second input port, 9 a first output port, 10 a second output port, 11 a liquid guide pipe, 12 a ring pipe, 13 a connecting pipe, 14 a sealing piece, 15 an electromagnet, 16 an electromagnetic switch, 17 a first pressure spring, 18 a sealing ring, 19 a single-phase baffle, 20 a fixing plate, 21 a culture dish, 22 a first motor, 23 an output pipe, 24 an observation window, 25 a take-out port, 26 a temporary storage tank, 27 a stirring device, 28 a second motor, 29 a stirring rod, 30 a stirring shaft, 31 a first magnet, 32 a second magnet, 33 a screw rod, 34 a fixing ring, 35 a third motor and 36 an electromagnetic valve.

Detailed Description

In order to better understand the technical content of the invention, specific embodiments are provided below, and the invention is further described with reference to the accompanying drawings.

Example 1

Referring to fig. 1 to 3, the stem cell separation and culture device provided by the invention comprises a separation cylinder 1 and a culture box 2, wherein a second inlet 8 for stem cell liquid to be separated is formed in the top of the separation cylinder 1, the separation cylinder 1 comprises a separation cavity 3 and a liquid storage cavity 4, the liquid storage cavity 4 is positioned below the separation cavity 3, a filter screen 5 is arranged in the separation cavity 3, the filter screen 5 can separate the stem cells from cleaning liquid, the stem cells are placed above the filter screen 5, the cleaning liquid flows into the bottom of the separation cavity 3, the separation cavity 3 is separated from the liquid storage cavity 4 through a separation piston 6, when the separation piston 6 moves downwards, a culture medium in the liquid storage cavity 4 can be pressed into the separation cavity 3 from a first inlet 7, the stem cells above the filter screen 5 are backflushed to separate the stem cells from the filter screen 5, the separation cavity 3 is communicated with a second outlet 10, and an output end of a liquid guide tube 11 is communicated with the culture box 2, the culture medium containing stem cells can flow into the culture box 2 from a second output port 10 through a liquid guide pipe 11, a sealing element 14 for closing or opening the second output port 10 is arranged above the separation cylinder 1, the sealing element 14 is in a closed state in a stem cell liquid separation stage, after the culture medium backflushs a filter screen 5 into the separation cavity 3, the sealing element 14 is moved upwards to enable the second output port 10 to be in an open state, the second output port 10 can be selected to be an annular output port, the culture medium containing stem cells is prevented from flowing to the liquid guide pipe 11 from the periphery of the separation cylinder 1, a first input port 7 is arranged on the side edge of the separation cavity 3, a first output port 9 is arranged at the bottom of the separation cavity 3, a single-phase baffle plate 19 which is opened inwards is arranged in the separation cavity 3, the single-phase baffle plate 19 is respectively matched with the first input port 7 and the first output port 9, and in the stem cell liquid separation stage, the single-, make first delivery outlet 9 be in the closed condition, impact single-phase baffle 19 when the culture medium and open first input port 7 after, single-phase baffle 19 blocks first delivery outlet 9, makes first delivery outlet 9 be in the closed condition, avoids culture medium and washing liquid to mix, and single-phase baffle 19 is integrative dual-purpose, the output of stock solution chamber 4 with first input port 7 union coupling, 2 internal rotation of incubator are connected with fixed plate 20, fixed plate 20 is fixed with a plurality of culture dishes 21, catheter 11 will contain the culture medium of stem cell a plurality of culture dishes 21.

The working principle is as follows: when the invention is used, the single-phase baffle plate 19 is positioned at the first input port 7, the first input port 7 is in a closed state, the sealing element 14 closes the second output port 10, the first output port 9 at the bottom of the separation cavity 3 is in an open state, stem cell liquid to be separated is poured into the separation cylinder 1 from the second input port 8, the filter screen 5 separates stem cells from stem cell cleaning liquid, the stem cells are placed above the filter screen 5, the cleaning liquid flows into the bottom of the separation cavity 3 through the filter screen 5 and the first output port 9 in sequence, after the stem cells are separated from the cleaning liquid, the separating piston 6 slides towards the bottom of the liquid storage cavity 4, the separating piston 6 presses the culture medium stored in the liquid storage cavity 4 into the first input port 7, the pressure at the outer side of the first input port 7 is greater than the pressure at the inner side, the single-phase baffle plate 19 rotates towards the separation cavity 3 and seals the first output port 9, the first output port 9 is in a closed state, the sealing element 14 is moved upwards at the same time, the second output port 10 is in an open state, the culture medium recoils on the filter screen 5 from the first input port 7, and stem cells placed above the filter screen 5 are led into the culture dish 21 from the second output port 10 through the liquid guide pipe 11 along with the culture medium, so that the separation and culture of the stem cells are realized.

Specifically, the sealing element 14 includes an electromagnet 15, an electromagnetic switch 16, a first pressure spring 17 and a sealing ring 18, the electromagnetic switch 16 is fixed above the separation cylinder 1, the first pressure spring 17 is disposed between the electromagnetic switch 16 and the electromagnet 15, the sealing ring 18 is connected to the electromagnet 15, and the sealing ring 18 is used for closing or opening the second output port 10. When the second output port 10 is in a closed state, the electromagnetic switch 16 is not powered on, the first pressure spring 17 springs the electromagnet 15 open, so that the sealing ring 18 moves downwards to seal the second output port 10, and when the electromagnetic switch 16 is powered on, the electromagnet 15 is attracted, so that the electromagnet 15 drives the sealing ring 18 to move upwards to open the second output port 10.

Specifically, the liquid guide tube 11 includes an annular tube 12 and a connecting tube 13, the annular tube 12 is disposed on the outer periphery of the separation chamber 3, the annular tube 12 is communicated with the second output port 10, the connecting tube 13 is communicated with an output end of the annular tube 12, and the connecting tube 13 is communicated with the incubator 2. The second outlet 10 is an annular outlet, and the culture medium containing the stem cells flows into the annular tube 12 from the second outlet 10 provided at the periphery of the separation chamber 3, is collected in the connecting tube 13, and is finally introduced into the incubator 2 through the connecting tube 13.

Example 2

Referring to fig. 1 to 5, the present embodiment is different from embodiment 1 in that a first motor 22 is provided at the bottom of the cultivation box 2, and an output shaft of the first motor 22 is connected to the fixing plate 20. The first motor 22 can drive the fixing plate 20 to rotate, so that the culture dish 21 fixed on the fixing plate 20 rotates to a position below the output end of the connecting pipe 13, thereby facilitating the receiving of the culture medium containing stem cells.

Specifically, the fixing plate 20 is a circular fixing plate 20, the culture dishes 21 are annularly fixed on the fixing plate 20, the end of the connecting pipe 13 is communicated with a plurality of output pipes 23 for guiding a culture medium containing stem cells into the culture dishes 21, and the output pipes 23 are fixed on the top of the incubator 2. The culture medium containing the stem cells may be simultaneously supplied to the plurality of culture dishes 21.

Specifically, the incubator 2 is provided with an observation window 24 and a take-out port 25 at the periphery thereof. The observation window 24 facilitates observation of the stem cell culture, and the removal port 25 facilitates insertion and removal of the culture dish 21.

Example 3

Referring to fig. 1 to 3, the present embodiment is different from embodiment 1 in that the catheter 11 is communicated with the culture chamber 2 through a temporary storage chamber 26, a stirring device 27 for uniformly distributing stem cells in the culture medium is arranged in the temporary storage chamber 26, the stirring device 27 includes a second motor 28 and a plurality of stirring rods 29, an output shaft of the second motor 28 is connected with a stirring shaft 30, and the plurality of stirring rods 29 are connected with the stirring shaft 30. The culture medium containing the stem cells is temporarily stored in the temporary storage box 26, the culture medium is stirred by the stirring rod 29, the stem cells are uniformly distributed in the culture medium, the rotating speed of the motor can be adjusted according to actual requirements, the material of the stirring rod 29 is selected, and the probability of damage to the stem cells is reduced, wherein the electromagnetic valve 36 is arranged between the temporary storage box 26 and the culture box 2.

Example 4

Referring to fig. 1 to 4, the present embodiment is different from embodiment 1 in that a first magnet 31 is disposed on the inner periphery of the partition piston 6, and a driving device (not shown) for moving the partition piston 6 up and down is disposed on the outer side of the separation cylinder 1. The separation piston 6 can move up and down between the separation cavity 3 and the liquid storage cavity 4 through the matching of a driving device (not shown) and the first magnet 31, so that the purposes of separation and stem cell culture are realized.

Specifically, drive arrangement (not marked) includes second magnet 32, lead screw 33, solid fixed ring 34 and third motor 35, second magnet 32 locates gu fixed ring 34's inside, gu fixed ring 34 sliding connection in the outside of cylinder 1, the output shaft of third motor 35 with lead screw 33 is connected, lead screw 33 with gu fixed ring 34 threaded connection, second magnet 32 with first magnet 31 is mutually supported. And starting a third motor 35, driving the fixing ring 34 and the second magnet 32 to move downwards by the screw rod 33 connected with the output shaft of the motor, so that the separating piston 6 moves downwards to extrude the culture medium in the liquid storage cavity 4, and the culture medium is pressed into the first input port 7.

The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.

Claims

1. A stem cell isolated culture device, characterized in that: comprises a separating cylinder and an incubator, wherein the separating cylinder comprises a separating cavity and a liquid storage cavity, a filter screen is arranged in the separating cavity, the separation cavity is separated from the liquid storage cavity by a separation piston and is communicated with a liquid guide pipe through a second output port, the output end of the liquid guide pipe is communicated with the incubator, a sealing piece used for closing or opening the second output port is arranged above the separating cylinder, a first input port is arranged on the side edge of the separation cavity, a first output port is arranged at the bottom of the separation cavity, the separation cavity is provided with a single-phase baffle plate which is opened inwards, the single-phase baffle plate is respectively matched with the first input port and the first output port, the output end of the liquid storage cavity is connected with the first input port pipe, the incubator is rotationally connected with a fixing plate, the fixed plate is fixed with a plurality of culture dishes, and the liquid guide pipe guides the culture medium containing the stem cells into the culture dishes.

2. The stem cell separation and culture device according to claim 1, wherein: the sealing piece comprises an electromagnet, an electromagnetic switch, a first pressure spring and a sealing ring, the electromagnetic switch is fixed above the separating cylinder, the first pressure spring is arranged between the electromagnetic switch and the electromagnet, the sealing ring is connected with the electromagnet, and the sealing ring is used for closing or opening the second output port.

3. The stem cell separation and culture device according to claim 1, wherein: the liquid guide pipe comprises an annular pipe and a connecting pipe, the annular pipe is arranged on the periphery of the outer portion of the separation cavity, the annular pipe is communicated with the second output port, the connecting pipe is communicated with the output end of the annular pipe, and the connecting pipe is communicated with the incubator.

4. The stem cell separation and culture device according to claim 1, wherein: the bottom of incubator is equipped with first motor, the output shaft of first motor with the fixed plate is connected.

5. The isolated stem cell culture apparatus of claim 4, wherein: the fixed plate is a round fixed plate, the culture dishes are fixed on the fixed plate in an annular mode, the tail ends of the connecting pipes are communicated with a plurality of output pipes used for guiding a culture medium containing stem cells into the culture dishes, and the output pipes are fixed at the top of the incubator.

6. The stem cell separation and culture device according to claim 5, wherein: the periphery of the incubator is provided with an observation window and a taking-out port.

7. The stem cell separation and culture device according to claim 1, wherein: the liquid guide pipe is communicated with the culture box through a temporary storage box, a stirring device used for enabling stem cells to be uniformly distributed in the culture medium is arranged in the temporary storage box, the stirring device comprises a second motor and a plurality of stirring rods, an output shaft of the second motor is connected with a stirring shaft, and the stirring rods are connected with the stirring shaft.

8. The stem cell separation and culture device according to claim 1, wherein: the inner periphery of the separation piston is provided with a first magnet, and the outer side of the separation cylinder is provided with a driving device which enables the separation piston to move up and down.

9. The stem cell separation and culture device according to claim 8, wherein: drive arrangement includes second magnet, lead screw, solid fixed ring and third motor, the second magnet is located gu fixed ring's inside, gu fixed ring sliding connection in the outside of cylinder, the output shaft of third motor with the lead screw is connected, the lead screw with gu fixed ring threaded connection, the second magnet with first magnet is mutually supported.