CN212102826U - Device for dispersing cells and quantitatively subpackaging cell sap - Google Patents
Device for dispersing cells and quantitatively subpackaging cell sap Download PDFInfo
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- CN212102826U CN212102826U CN202020227594.0U CN202020227594U CN212102826U CN 212102826 U CN212102826 U CN 212102826U CN 202020227594 U CN202020227594 U CN 202020227594U CN 212102826 U CN212102826 U CN 212102826U
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Abstract
The utility model discloses a device for cell dispersion and quantitative cell sap subpackage, a barrel cover is fixed on the top of a cell dispersion barrel, an air respirator, a liquid inlet interface and a liquid outlet interface are fixed on the top of the barrel cover, a dispersion pipe is fixed on the bottom end of the liquid inlet interface, an extension pipe is fixed on the bottom end of the liquid outlet interface, the device also comprises a three-way valve, a quantitative subpackage peristaltic pump and a circulating peristaltic pump, the top end of the liquid outlet interface, the liquid inlet end of the quantitative subpackage peristaltic pump and the liquid inlet end of the circulating peristaltic pump are respectively connected with one end of the three-way valve through connecting pipes, the liquid outlet end of the circulating peristaltic pump is connected with the top end of the liquid inlet interface through connecting pipes, the liquid outlet end of the quantitative subpackage peristaltic pump is connected with a subpackaging device through connecting pipes, a branch pipe is also arranged on the connecting pipe between the circulating peristaltic pump and the three, the utility model discloses cell dispersion effect is good, cell passage is efficient, easy operation, pollution risk are little.
Description
Technical Field
The utility model relates to a device that is used for cell dispersion and cell sap ration partial shipment.
Background
In the traditional rotary bottle culture process, after pancreas is added into a cell bottle, after cells are digested from the bottle wall, cell suspension is added, a bottle cover is covered, an operator holds the cell bottle by two hands, and shakes the cell bottle violently to digest the cells; or after adding the suspension, the dispersed cells were blown through a pipette. After one vial of cells was dispersed as above, a second vial of cells was run in the same manner. Thus, the labor intensity is high when all the cell bottles are repeatedly performed. Moreover, the serum contained in the cell sap is likely to destroy a large number of cells by vigorous shaking. After the cells are well dispersed, according to the passage ratio (assumed to be 1:3 passage in the expression, each bottle is filled with 400ml of liquid finally), each mother bottle is filled with 1200ml of culture solution by manual pouring, the mother bottle is sealed, the cells are uniformly mixed by shaking, and then the cells are poured into 3 finally cultured cell bottles on average. When liquid is poured manually, the liquid volume is determined according to the scales of the rotary bottle, the error is large, the number of cells added into each cell bottle is inconsistent, and the cell consistency is poor. The traditional method has the defects of extensive production operation, poor cell uniformity, large cell loss, large action range of personnel during operation, high pollution risk of an operation mode of transferring liquid from a bottle to the bottle, and unfavorable improvement on production efficiency and vaccine quality.
The glass rotary bottle needs to be cleaned and sterilized repeatedly when being used repeatedly, the labor capacity is large, and cells on the rotary bottle after being used for many times often have poor growth vigor due to the fact that the inner surface of the rotary bottle is not cleaned in place or the inner surface of the rotary bottle is scratched when being cleaned, and production is directly influenced. Based on the problems, enterprises begin to use disposable cell culture bottles which are made of PP materials and are disposable, the problem of repeated use does not exist, and the disposable cell culture bottles are more favorable for cell growth than traditional glass rotary bottles, but the cells grow well and are firmly adhered to the bottles, and cell masses digested by pancreatin are difficult to disperse by adopting two common traditional dispersing processes such as a mode of violently shaking the cell bottles or a mode of blowing and beating by suction pipes, so that the wide application of the disposable cell culture bottles in large production is limited.
SUMMERY OF THE UTILITY MODEL
The utility model aims to overcome the defects of the prior art and provide a device for dispersing cells and quantitatively subpackaging cell sap.
The technical scheme of the utility model is that: a device for cell dispersion and quantitative cell sap subpackaging comprises a cell dispersion barrel and a plurality of cell bottles, wherein a barrel cover is fixed at the top of the cell dispersion barrel, an air respirator, a liquid inlet interface and a liquid outlet interface are fixed at the top of the barrel cover, a dispersion pipe is fixed at the bottom end of the liquid inlet interface, penetrates through the bottom of the barrel cover and is a stainless steel pipe body with a thick upper part and a thin lower part, micropores are densely distributed at the bottom end of the dispersion pipe, an extension pipe is fixed at the bottom end of the liquid outlet interface, the bottom of the extension pipe is inserted into the cell dispersion barrel and is abutted against the bottom of the cell dispersion barrel, the device further comprises a three-way valve, a quantitative subpackaging peristaltic pump and a circulating peristaltic pump, the top end of the liquid outlet interface, the liquid inlet end of the quantitative subpackaging peristaltic pump and the liquid inlet end of the circulating peristaltic pump are respectively connected with one end of the three-way valve through connecting pipes, and the liquid outlet end of, the liquid outlet end of the quantitative subpackaging peristaltic pump is connected with the subpackaging device through a connecting pipe, a branch pipe is further arranged on the connecting pipe between the circulating peristaltic pump and the three-way valve, a liquid sucking device is connected to the end portion of the branch pipe, and a valve is further arranged on the branch pipe.
Furthermore, the inner diameter of the lower end of the dispersion pipe is 0.1cm, the length of the dispersion pipe is about 20cm from the bottom to the top, small holes with the size of 0.5-1mm are punched on the section of the pipe, and the distance between the holes is about 5 mm.
Furthermore, a bottle cap is fixed on the cell bottle, a sub-packaging device interface, a liquid suction device interface and a ventilation opening are arranged on the bottle cap, and sealing covers are buckled on the sub-packaging device interface, the liquid suction device interface and the ventilation opening.
Furthermore, the connecting pipe is a silicone tube.
The utility model has the advantages that:
the utility model discloses when carrying out the cell dispersion, silicone tube cooperation dispersion tube clearance shrink under the action of circulating peristaltic pump shears the cell block piece in the liquid, and the cell is spouted by each utmost point pore of dispersion tube under the high pressure effect simultaneously and is extruded, can effectively disperse the firm cell block piece that bonds, and the cell dispersion is effectual.
The utility model is simple in operation, because there is not long-time uncovered operation, the cell passage operation pollution risk has obtained very big reduction.
The quantitative peristaltic pump is adopted to determine the subpackage volume for moving out or in the liquid from the bottle, the quantification is accurate, and the pollution risk is small.
Compared with the traditional passage of 1:3, the utility model can disperse more next generation cell suspension liquid with more bottles at one time, and the efficiency is greatly improved. And all the mother cells are collected in a barrel and then quantitatively subpackaged in cell bottles, and because the number of the cells in each bottle is consistent, the uniformity of the cell growth in each bottle is ensured, and the cells in the later period are convenient to use.
Drawings
Fig. 1 is a schematic structural diagram of the present invention;
fig. 2 is a plan view of the bottle cap of the present invention;
fig. 3 is a schematic structural diagram of the bottom end of the dispersion pipe of the present invention.
Detailed Description
The technical solution of the present invention is further specifically described below by way of examples and with reference to the accompanying drawings.
With reference to fig. 1 and 3, a device for cell dispersion and quantitative cell sap split charging includes a cell dispersion barrel 1 and a plurality of cell bottles 2, the cell dispersion barrel 1 is a transparent PP barrel with a volume of 20-50L, a barrel cover 3 is fixed on the top of the cell dispersion barrel 1, an air respirator 4, a liquid inlet port 5 and a liquid outlet port 6 are fixed on the top of the barrel cover 3, the bottom end of the liquid inlet port 5 penetrates through the bottom of the barrel cover 3 and is fixed with a dispersion pipe 7, the dispersion pipe 7 is a stainless steel tube with a thick upper part and a thin lower part, the bottom end of the dispersion pipe 7 is densely distributed with fine pores, the inner diameter of the lower end of the dispersion pipe is 0.1cm, the bottom end of the liquid outlet port 6 is fixed with an extension pipe 8, and the bottom of the extension pipe 8 is inserted into the cell dispersion barrel 1 and abutted against the bottom of the cell dispersion barrel 1;
the quantitative subpackaging device is characterized by further comprising a three-way valve 9, a quantitative subpackaging peristaltic pump 10 and a circulating peristaltic pump 11, wherein the top end of the liquid outlet interface 6, the liquid inlet end of the quantitative subpackaging peristaltic pump 10 and the liquid inlet end of the circulating peristaltic pump 11 are respectively connected with one end of the three-way valve 9 through silicone tubes, the liquid outlet end of the circulating peristaltic pump 11 is connected with the top end of the liquid inlet interface 5 through a silicone tube, and the liquid outlet end of the quantitative subpackaging peristaltic pump 10 is connected with the subpackaging device 12 through a silicone tube;
a branch pipe 13 is further arranged on the silicone tube between the circulating peristaltic pump 11 and the three-way valve 9, a liquid suction device 14 is connected to the end of the branch pipe 13, and a valve 15 is further arranged on the branch pipe 13;
as shown in fig. 2, a cap 16 is fixed to the cell bottle 2, the cap 16 has a dispenser interface 17, a pipette interface 18, and a vent 19, and in a state where liquid is not sucked and dispensed, the dispenser interface 17, the pipette interface 18, and the vent 19 are all fitted with a sealing cap.
The utility model discloses an application step is:
step one, adding a cell suspension into the cell bottle 2: the cell bottle 2 after cell digestion is placed on an operation table after surface disinfection, a three-way valve 9 is adjusted to enable a cell dispersing barrel 1 and a quantitative subpackaging peristaltic pump 10 to be communicated and disconnect the cell dispersing barrel 1 and a circulating peristaltic pump 11, the quantitative subpackaging peristaltic pump 10 is started, a subpackaging device 12 is sequentially connected with a subpackaging device interface 17 of each cell bottle 2, and cell suspension liquid is injected into each cell bottle 2. (in the process of injection, the sealing cover on the ventilation opening 19 is opened, and the injection is finished and then the sealing cover is buckled.)
Step two, collecting the cell suspension in the cell bottle 2 to a cell dispersion barrel 1: gently rotating the cell bottle 2, washing the cells into the suspension, sterilizing the surface of the cell rotating bottle, and then placing the cell rotating bottle on an operation table; keeping the disconnection state of the cell dispersing barrel 1 and the circulating peristaltic pump 11, opening the valve 15 on the branch pipe 13, starting the circulating peristaltic pump 11, connecting the pipette 14 with the pipette connector 18 of each cell bottle 2, and pumping the cell suspension from each cell bottle 2 into the cell dispersing barrel 1. (in the process of drawing liquid, the sealing cover on the ventilating opening 19 is opened and then buckled after drawing is finished.)
Step three, dispersing cells: adjusting the three-way valve 9 to communicate the cell dispersing barrel 1 with the circulating peristaltic pump 11, disconnecting the cell dispersing barrel 1 from the quantitative subpackaging peristaltic pump 10, closing the valve 15 on the branch pipe 13, and circulating the cell suspension in the cell dispersing barrel 1 through the circulating peristaltic pump 11; in the circulation process, the silicone tube shrinks at intervals under the action of the circulating peristaltic pump 11 to cut the cells which are bonded into clusters or sheets, so that the dispersion efficiency is improved, after the cell suspension enters the dispersion tube 7, the cells are sprayed and extruded from each extremely fine hole of the dispersion tube under the action of high pressure, and the cell clusters are successfully dispersed after a certain period of circulation; wherein, the rotating speed of the circulating peristaltic pump is set to 3000r/min, when the total cell suspension is less than or equal to 10000ml, the dispersion time is 35-40min, and when the total cell suspension is more than 10000ml, the dispersion time is 45-55 min.
Step four, quantitative subpackaging: after the dispersion ends, the operation as the step one is carried out, and the cell sap is quantitatively dispensed into the cell bottle 2 by using the quantitative dispensing peristaltic pump 10.
It will be understood that when an element is referred to as being "secured to" another element, it can be directly on the other element or intervening elements may also be present. When an element is referred to as being "connected" to another element, it can be directly connected to the other element or intervening elements may also be present. The terms "vertical," "horizontal," "left," "right," and the like as used herein are for illustrative purposes only and do not represent the only embodiments.
Unless defined otherwise, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. The terminology used herein in the description of the invention is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. As used herein, the term "and/or" includes any and all combinations of one or more of the associated listed items.
The above description is only a preferred embodiment of the present invention, and should not be taken as limiting the invention, and any modifications, equivalent replacements, improvements, etc. made within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (4)
1. The utility model provides a device that is used for cell dispersion and cell sap ration partial shipment, includes a plurality of cell bottles which characterized in that: the cell dispersing device is characterized by further comprising a cell dispersing barrel, a barrel cover is fixed to the top of the cell dispersing barrel, an air respirator, a liquid inlet interface and a liquid outlet interface are fixed to the top of the barrel cover, a dispersing pipe is fixed to the bottom of the liquid inlet interface, the bottom of the liquid inlet interface penetrates through the bottom of the barrel cover and is fixed to the bottom of the barrel cover, the dispersing pipe is a stainless steel pipe body with a thick upper portion and a thin lower portion, extremely fine holes are densely distributed in the bottom of the dispersing pipe, an extension pipe is fixed to the bottom of the liquid outlet interface, the bottom of the extension pipe is inserted into the cell dispersing barrel and is abutted to the bottom of the cell dispersing barrel, the cell dispersing device further comprises a three-way valve, a quantitative subpackaging peristaltic pump and a circulating peristaltic pump, the top end of the liquid outlet interface, the liquid inlet end of the quantitative subpackaging peristaltic pump and the liquid inlet end of the circulating peristaltic pump are respectively connected with one end of the three, a branch pipe is further arranged on a connecting pipe between the circulating peristaltic pump and the three-way valve, a liquid suction device is connected to the end of the branch pipe, and a valve is further arranged on the branch pipe.
2. The apparatus according to claim 1, wherein the apparatus comprises: the inner diameter of the lower end of the dispersion pipe is 0.1 cm.
3. The apparatus according to claim 1, wherein the apparatus comprises: the cell bottle is fixed with a bottle cap, the bottle cap is provided with a dispenser interface, a liquid sucker interface and a ventilation opening, and the dispenser interface, the liquid sucker interface and the ventilation opening are all buckled with sealing covers.
4. The apparatus according to claim 1, wherein the apparatus comprises: the connecting pipe is a silicone tube.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202020227594.0U CN212102826U (en) | 2020-02-28 | 2020-02-28 | Device for dispersing cells and quantitatively subpackaging cell sap |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202020227594.0U CN212102826U (en) | 2020-02-28 | 2020-02-28 | Device for dispersing cells and quantitatively subpackaging cell sap |
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| CN212102826U true CN212102826U (en) | 2020-12-08 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113697146A (en) * | 2021-08-20 | 2021-11-26 | 苏州博腾生物制药有限公司 | Cell subpackaging method in cell bank building process |
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2020
- 2020-02-28 CN CN202020227594.0U patent/CN212102826U/en active Active
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN113697146A (en) * | 2021-08-20 | 2021-11-26 | 苏州博腾生物制药有限公司 | Cell subpackaging method in cell bank building process |
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