CN212025374U - Novel inoculation device for three-dimensional cell culture - Google Patents
Novel inoculation device for three-dimensional cell culture Download PDFInfo
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- CN212025374U CN212025374U CN202020330752.5U CN202020330752U CN212025374U CN 212025374 U CN212025374 U CN 212025374U CN 202020330752 U CN202020330752 U CN 202020330752U CN 212025374 U CN212025374 U CN 212025374U
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Abstract
The utility model relates to a cell culture, regenerative medicine technique and environmental health research field, concretely relates to a novel inoculation device for three-dimensional cultivation of cell, its characterized in that: including the stock solution bottle, the screen cloth, filler and storage pipe, be equipped with the storage chamber at both ends about running through in the storage pipe, both ends aperture is unanimous about the storage pipe, the equal detachable in both ends is connected with the sleeve pipe about the storage pipe, be equipped with the screen cloth between two sleeve pipes and the storage pipe, it has the filler to fill between two screen cloths in the storage pipe, the one end detachable of keeping away from the storage pipe on the sleeve pipe is connected with the stock solution bottle, set up the ascending open stock solution chamber in the stock solution bottle, the stock solution bottle is linked together through sleeve pipe and storage pipe. The utility model has the advantages of simple and practical overall structure, the simple operation is convenient for realize the three-dimensional cultivation of cell, can avoid contacting many times with the air at the inoculation in-process, ensures simultaneously that the porous microballon in the storage pipe carries out abundant even inoculation to the cell.
Description
Technical Field
The utility model relates to a cell culture, regenerative medicine technique and environmental health research field, specificly relate to a novel inoculation device for three-dimensional cultivation of cell.
Background
In the in vitro cell culture technology, the traditional two-dimensional cell culture has the advantages of simple culture, easy operation, low price and the like, and currently occupies the mainstream position, but the characteristics of proliferation, differentiation, gene expression and the like of a cell line cultured in vitro are greatly different from those of primary cells and tissues in vivo due to the in vitro survival selection pressure. However, for most of microcarriers, the traditional cell inoculation method is to blend cells and microcarriers, so that the cells can only carry out adhesion growth on the outer surface of the microcarriers, and even if the microcarriers are porous microspheres, the cells only adhere to the outer pore surfaces of the microcarriers.
To solve this problem, utility model patent publication No. CN 209923357U provides an inoculating device for three-dimensional cell culture. The utility model provides an inoculation device for three-dimensional cell culture, which comprises a tubular hollow main body, a rubber plug, a sieve plate, a screen and a filler; the upper end and the lower end of the tubular hollow main body are provided with openings, and the opening at the lower end is smaller than the opening at the upper end; the rubber plug is plugged in an upper end opening of the tubular hollow main body; the sieve plate is positioned below the rubber plug and is horizontally fixed in the tubular hollow main body; the screen is positioned at the lower end opening of the tubular hollow main body; the filler is filled between the sieve plate and the sieve mesh, however, in the cell inoculation process of the device, the effluent flowing out from the lower end of the device needs to be collected by a culture dish, and then is added into the upper end opening of the device, and the process is repeated for a plurality of times, so that the cell suspension can be contacted with the air for a plurality of times in the inoculation process, and in addition, the cell suspension is always added from the upper end opening of the device, so that the cell inoculation of the porous microspheres at the lower part of the device is insufficient, and the cell inoculation of the porous microspheres in the device is uneven.
SUMMERY OF THE UTILITY MODEL
The to-be-solved technical problem of the utility model is to provide an overall structure is simple, practical, and the simple operation is convenient for realize the three-dimensional cultivation of cell, can avoid contacting many times with the air at the inoculation in-process, ensures simultaneously that the porous microsphere in the storage pipe carries out the novel inoculation device that is used for three-dimensional cultivation of cell of abundant even inoculation to the cell.
In order to solve the technical problem, the utility model discloses a technical scheme as follows: the utility model provides a novel inoculation device for three-dimensional cultivation of cell, including the stock solution bottle, the screen cloth, filler and storage pipe, be equipped with the storage chamber that runs through both ends from top to bottom in the storage pipe, both ends aperture is unanimous about the storage pipe, the equal detachable in both ends is connected with the sleeve pipe about the storage pipe, be equipped with the screen cloth between two sleeve pipes and the storage pipe, it has the filler to fill between two screen cloths in the storage pipe, the one end detachable who keeps away from the storage pipe on the sleeve pipe is connected with the stock solution bottle, it upwards opens the stock solution chamber to have seted up in the stock solution bottle, the stock solution bottle.
Further, be threaded connection between storage pipe and the sleeve pipe and between sleeve pipe and the stock solution bottle, all be equipped with sealed pad between storage pipe and the sleeve pipe and between sleeve pipe and the stock solution bottle.
Furthermore, the storage pipe is in a shape that the middle is thin and the two ends are thick.
Furthermore, the liquid storage bottle is transparent, and the bottom surface of the liquid storage bottle is a plane.
Furthermore, this novel inoculation device still includes supporting mechanism, and supporting mechanism is used for playing the supporting role and making the storage pipe be vertical layout to the storage pipe.
Further, the supporting mechanism includes the base, fixed block and movable block, the base includes bottom plate and backup pad, the backup pad is two and parallel fixed locates the bottom plate upper end, the fixed block is located between two backup pads, the fixed block both ends are all fixed and are equipped with the fixed axle, the fixed axle passes through the rotatable backup pad of locating of bearing, all set up flutedly on movable block and the fixed block, two recesses merge and be formed with the fixed orifices with storage pipe middle part shape looks adaptation, movable block and fixed block bolted connection and the tight fixed storage pipe of clamp, still be equipped with the stop gear who is used for playing limiting displacement to the rotation of fixed axle in the backup pad of fixed axle and corresponding one side.
Further, stop gear includes the spacer pin, and backup pad lateral surface upper portion is fixed and is equipped with the stopper, has seted up spacing hole on the stopper, and the fixed axle tip is seted up with spacing hole from top to bottom the through-hole that aligns, and the separable inserting of spacer pin is located through-hole and spacing downthehole.
Further, the aperture of the screen is smaller than the particle size of the filler and can allow the cells and the cell culture medium to pass through, and the aperture size of the screen is 50-100 um.
Furthermore, the filler is porous microspheres, the particle size of the porous microspheres is 100-300um, and the pore size of the pores on the porous microspheres is 20-50 um.
Furthermore, the porous microspheres are L-polylactic acid porous microspheres.
As can be seen from the above description, the utility model provides a novel inoculation device for three-dimensional cell culture, the overall structure is simple, practical, convenient to use, through the principle of applying gel chromatography, cells can be inoculated inside porous microspheres, the three-dimensional cell culture is realized, through arranging the liquid storage bottles at the two ends of the storage tube, in the cell inoculation process, no additional culture dish is needed to collect the effluent liquid, the liquid storage bottle at the lower end can collect the effluent liquid, after the collection is completed, the storage tube is rotated, the liquid storage bottle with the effluent liquid is rotated to the upper end, thereby the smooth proceeding of the cell inoculation process is ensured, the whole cell inoculation process does not need to hold the storage tube by hand, the operation is easier and more convenient, through adopting the arrangement, the cell inoculation process is all in the closed space, the multiple contact with the air can be avoided, in addition, in the inoculation process of cells, the storage tube is overturned for multiple times, so that the porous microspheres in the liquid storage bottle can be ensured to be fully and uniformly inoculated to the cells.
Drawings
Fig. 1 is a schematic view of the three-dimensional structure of the novel inoculation device for three-dimensional cell culture of the present invention.
Fig. 2 is an exploded perspective view of fig. 1.
Fig. 3 is a partially enlarged schematic view of a portion a in fig. 2.
Fig. 4 is a partially enlarged schematic view of a portion B in fig. 2.
Fig. 5 is a left side view of fig. 1.
Fig. 6 is a schematic cross-sectional view of aspect C-C of fig. 5.
Fig. 7 is a partially enlarged view of fig. 6 at D.
Fig. 8 is a schematic diagram of the state that the novel inoculation device for three-dimensional cell culture of the present invention is filled with filler.
In the figure: 1-liquid storage bottle; 11-a liquid storage cavity; 2-screening a screen; 3-a filler; 4-a storage pipe; 41-a material storage cavity; 5-sleeving a pipe; 51-annular bumps; 61-a first thread turn; 62-a second thread turn; 7-a sealing gasket; 8-a support mechanism; 81-base; 811-a backplane; 812-a support plate; 82-fixing blocks; 83-a movable block; 84-a fixed shaft; 841-a via hole; 85-a bearing; 86-fixing holes; 87-a limit pin; 88-a limiting block; 881 limit holes.
Detailed Description
The present invention will be further described with reference to the following detailed description.
As shown in fig. 1, fig. 2, fig. 5, fig. 6 and fig. 8, a novel inoculation device for three-dimensional cell culture, including stock solution bottle 1, screen cloth 2, filler 3 and storage pipe 4, be equipped with the storage cavity 41 that runs through both ends from top to bottom in the storage pipe 4, the both ends open pore diameter is unanimous about storage pipe 4, the equal detachable of both ends is connected with sleeve pipe 5 about storage pipe 4, two sleeve pipe 5 with be equipped with between the storage pipe 4 screen cloth 2, in two in the storage pipe 4 it has filler 3 to fill between the screen cloth 2, the sleeve pipe 5 is gone up in keeping away from the one end detachable of storage pipe 4 is connected with stock solution bottle 1, upwards open stock solution chamber 11 has been seted up in the stock solution bottle 1, stock solution bottle 1 pass through sleeve pipe 5 with the storage pipe 4 is linked together.
As shown in fig. 3 and 7, the storage tube 4 and the sleeve 5 and the liquid storage bottle 1 are both in threaded connection, in this embodiment, a first thread ring 61 is fixedly arranged at an opening of the liquid storage bottle 1, a second thread ring 62 is fixedly arranged at openings at two ends of the sleeve 5, external threads are arranged on outer surfaces of the first thread ring 61 and the second thread ring 62, internal threads adapted to the external threads are arranged on inner surfaces of openings at two ends of the sleeve 5, so as to facilitate detachable connection between the storage tube 4 and the sleeve 5 and between the sleeve 5 and the liquid storage bottle 1, in order to facilitate fixing the screen 2, an annular convex block 51 is fixedly arranged in the sleeve 5, the screen 2 is located between the annular convex block 51 and the end of the discharge tube, and sealing gaskets 7 are arranged between the storage tube 4 and the sleeve 5 and between the sleeve 5 and the liquid storage bottle 1, the sealing effect of the joint is ensured, and leakage is avoided.
As shown in fig. 6 and 8, the storage tube 4 is thin in the middle and thick at two ends, the liquid storage bottle 1 is transparent, so that the dripping condition of the cell suspension in the liquid storage bottle 1 can be observed conveniently, the bottom surface of the liquid storage bottle 1 is a plane, and the liquid storage bottle 1 can be placed conveniently when being taken down.
As shown in fig. 1, 2 and 4, the novel inoculation device further comprises a supporting mechanism 8, the supporting mechanism 8 is used for supporting the storage tube 4 and enabling the storage tube 4 to be vertically arranged, the supporting mechanism 8 comprises a base 81, a fixed block 82 and a movable block 83, the base 81 comprises a bottom plate 811 and two supporting plates 812, the two supporting plates 812 are parallelly and fixedly arranged at the upper end of the bottom plate 811, the fixed block 82 is arranged between the two supporting plates 812, two ends of the fixed block 82 are respectively and fixedly provided with a fixed shaft 84, the fixed shafts 84 are rotatably arranged on the supporting plates 812 through bearings 85, the movable block 83 and the fixed block 82 are respectively provided with a groove, the two grooves are combined to form a fixed hole 86 matched with the middle shape of the storage tube 4, the movable block 83 is in bolted connection with the fixed block 82 and clamps and fixes the storage tube 4, the fixing shaft 84 and the supporting plate 812 corresponding to one side of the fixing shaft 84 are further provided with a limiting mechanism for limiting rotation of the fixing shaft 84, and due to the arrangement of the supporting mechanism 8, the cell inoculation process is more labor-saving and easy, the storage pipe 4 does not need to be held by hands, and the cell inoculation process can be smoothly performed by rotating the storage pipe 4.
As shown in fig. 4, the limiting mechanism includes a limiting pin 87, a limiting block 88 is fixedly arranged on the upper portion of the outer side surface of the supporting plate 812, a limiting hole 881 is formed in the limiting block 88, a through hole 841 aligned with the limiting hole 881 is formed in the end portion of the fixing shaft 84, the limiting pin 87 is detachably inserted into the through hole 841 and the limiting hole 881, the structure is simple, practical and reliable, when the limiting pin 87 is taken out, the storage pipe 4 can be rotated and the upper and lower ends can be exchanged, after the rotation is completed, when the limiting pin 87 is inserted, the storage pipe 4 can be ensured to be stably in a vertical state, and the cell inoculation process can be smoothly performed.
The aperture of screen cloth 2 is less than the particle diameter of filler 3 and can make cell and cell culture medium pass through, the aperture size of screen cloth 2 is 50-100um, and preferred the aperture size of screen cloth 2 is 75 um.
The filler 3 is porous microspheres, the particle size of the porous microspheres is 100-300um, the pore size of the pores on the porous microspheres is 20-50um, and the porous microspheres are L-polylactic acid porous microspheres.
By applying the principle of gel chromatography, cells can be inoculated in the porous microspheres to realize the three-dimensional culture of the cells, the two ends of the storage tube 4 are provided with the storage bottles 1, in the process of cell inoculation, no other culture dish is needed to collect effluent, the effluent can be collected by the liquid storage bottle 1 at the lower end, after the collection is finished, then the storage tube 4 is rotated to ensure that the liquid storage bottle 1 filled with effluent liquid rotates to the upper end, thereby ensuring the smooth proceeding of the cell inoculation process, the whole cell inoculation process does not need to hold the storage tube 4 by hand, the operation is easier and more convenient, by adopting the arrangement, thereby ensuring that the cell inoculation process is in a closed space and can avoid multiple contact with air, in addition, in the process of inoculating cells, the storage tube 4 is overturned for multiple times, so that the porous microspheres in the storage bottle 1 can be ensured to sufficiently and uniformly inoculate the cells.
The use method of the novel inoculation device for three-dimensional cell culture comprises the following steps: when in use, the device is sterilized and placed on a super clean bench, firstly, a storage pipe 4 is fixedly arranged on a fixed block 82 through a movable block 83, the storage pipe 4 is vertically arranged, a culture medium is added from the upper end of the storage pipe 4 for infiltration, a culture dish is placed below the storage pipe 4 to collect the flowing culture medium, repeated rinsing is carried out for 3-5 times, the culture medium is added for the first time, the opening at the lower end of the storage pipe 4 can be plugged and infiltrated for 5-10 min to ensure that a filler 3 is in a wet state, the prepared cell suspension is added into one storage bottle, the storage bottle filled with the cell suspension is arranged at the lower end of the storage pipe 4, then, the other empty storage bottle is arranged at the upper end of the storage pipe 4, at the moment, a limiting pin 87 is taken out, the storage pipe 4 is turned over, so that the upper end and the lower end of the storage pipe 4 are exchanged, then the limit pin 87 is inserted again for fixing, at this time, the storage bottle filled with the cell suspension is positioned at the upper end of the storage tube 4, the storage bottle is kept in a standing state, the cell suspension flows through the storage tube 4 under the action of gravity and flows into the storage bottle 1 at the lower end, after the collection is completed, the limit pin 87 is taken out again, and the storage pipe 4 is turned over, so that the upper end and the lower end of the storage pipe 4 are exchanged again, then the limit pin 87 is inserted again for fixing, the cell suspension flows through the storage tube 4 again under the action of gravity and flows into the liquid storage bottle 1 at the lower end, the operation is repeated for 3-5 times, then, the liquid storage bottle 1 with the effluent liquid at the lower end of the storage pipe 4 is taken down, the sleeve 5 at the lower end of the storage pipe 4 is taken down, so that the screen 2 is taken down, then, the filler 3 in the storage tube 4 is gently flushed into the culture dish by using the culture medium for culture.
The aforesaid is only a plurality of concrete implementation manners of the utility model, nevertheless the utility model discloses a design concept is not limited to this, and the ordinary use of this design is right the utility model discloses carry out immaterial change, all should belong to the act of infringement the protection scope of the utility model.
Claims (10)
1. A novel inoculation device for three-dimensional cell culture is characterized in that: including stock solution bottle, screen cloth, filler and storage pipe, be equipped with the storage chamber that runs through both ends from top to bottom in the storage pipe, both ends aperture diameter is unanimous about the storage pipe, the equal detachable in both ends is connected with the sleeve pipe about the storage pipe, two the sleeve pipe with be equipped with between the storage pipe the screen cloth, storage intraductal in two it has the filler to fill between the screen cloth, on the sleeve pipe in keeping away from the one end detachable of storage pipe is connected with the stock solution bottle, upwards open stock solution chamber has been seted up in the stock solution bottle, the stock solution bottle passes through the sleeve pipe with the storage pipe is linked together.
2. A novel inoculating device for three-dimensional culture of cells as claimed in claim 1, wherein: the storage tube with be threaded connection between the sleeve pipe and between the sleeve pipe with the stock solution bottle, the storage tube with between the sleeve pipe and the sleeve pipe with all be equipped with sealed pad between the stock solution bottle.
3. A novel inoculating device for three-dimensional culture of cells as claimed in claim 1, wherein: the storage pipe is in a shape that the middle is thin and the two ends are thick.
4. A novel inoculating device for three-dimensional culture of cells as claimed in claim 1, wherein: the liquid storage bottle is transparent, and the bottom surface of the liquid storage bottle is a plane.
5. A novel inoculating device for three-dimensional culture of cells as claimed in claim 1, wherein: the novel inoculation device further comprises a supporting mechanism, and the supporting mechanism is used for supporting the storage pipe and enabling the storage pipe to be vertically arranged.
6. A novel inoculation device for three-dimensional cell culture according to claim 5, characterized in that: the supporting mechanism comprises a base, a fixed block and a movable block, the base comprises a bottom plate and a supporting plate, the supporting plate is two and is parallelly fixed to be located the bottom plate upper end, the fixed block is located two between the supporting plate, the fixed block both ends are all fixed and are provided with the fixed shaft, the fixed shaft passes through the rotatable locating of bearing in the supporting plate, the movable block with all set up flutedly on the fixed block, two the recess merge be formed with the fixed orifices of storage pipe middle part shape looks adaptation, the movable block with fixed block bolted connection presss from both sides tightly fixedly the storage pipe, fixed shaft and corresponding one side still be equipped with in the supporting plate be used for right the rotation of fixed shaft plays limiting displacement's stop gear.
7. A novel inoculation device for three-dimensional cell culture according to claim 6, characterized in that: the limiting mechanism comprises a limiting pin, a limiting block is fixedly arranged on the upper portion of the outer side face of the supporting plate, a limiting hole is formed in the limiting block, a through hole which is aligned with the limiting hole vertically is formed in the end portion of the fixing shaft, and the limiting pin is detachably inserted into the through hole and the limiting hole.
8. A novel inoculating device for three-dimensional culture of cells as claimed in claim 1, wherein: the aperture of the screen is smaller than the particle size of the filler and can allow cells and cell culture medium to pass through, and the size of the aperture of the screen is 50-100 um.
9. A novel inoculating device for three-dimensional culture of cells as claimed in claim 1, wherein: the filler is porous microspheres, the particle size of the porous microspheres is 100-300um, and the pore size of pores on the porous microspheres is 20-50 um.
10. A novel inoculating device for three-dimensional culture of cells as claimed in claim 9, wherein: the porous microspheres are L-polylactic acid porous microspheres.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202020330752.5U CN212025374U (en) | 2020-03-17 | 2020-03-17 | Novel inoculation device for three-dimensional cell culture |
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| Application Number | Priority Date | Filing Date | Title |
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| CN202020330752.5U CN212025374U (en) | 2020-03-17 | 2020-03-17 | Novel inoculation device for three-dimensional cell culture |
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| CN212025374U true CN212025374U (en) | 2020-11-27 |
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| CN202020330752.5U Active CN212025374U (en) | 2020-03-17 | 2020-03-17 | Novel inoculation device for three-dimensional cell culture |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112592812A (en) * | 2020-12-15 | 2021-04-02 | 山东省农业科学院畜牧兽医研究所 | Inoculation device for muscle cell culture |
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2020
- 2020-03-17 CN CN202020330752.5U patent/CN212025374U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112592812A (en) * | 2020-12-15 | 2021-04-02 | 山东省农业科学院畜牧兽医研究所 | Inoculation device for muscle cell culture |
| CN112592812B (en) * | 2020-12-15 | 2022-08-02 | 山东省农业科学院畜牧兽医研究所 | Inoculation device for muscle cell culture |
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