CN222099996U - A cell culture plate with a filter device for chondrogenic differentiation of stem cells - Google Patents
A cell culture plate with a filter device for chondrogenic differentiation of stem cells Download PDFInfo
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- CN222099996U CN222099996U CN202420664189.3U CN202420664189U CN222099996U CN 222099996 U CN222099996 U CN 222099996U CN 202420664189 U CN202420664189 U CN 202420664189U CN 222099996 U CN222099996 U CN 222099996U
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Abstract
A cell culture plate with a filtering device for stem cell chondrogenic differentiation relates to the technical field of cell culture containers and solves the problems that in the prior art, liquid exchange operation is difficult, newly formed cartilage balls are easily damaged by touching of a pipette tip, or liquid exchange is incomplete, and the growth state of cells is affected. A cell culture plate with a filtering device for chondrogenic differentiation of stem cells comprises a bottom plate and a plate cover detachably connected to the bottom plate, wherein a plurality of culture holes are formed in the bottom plate, a plastic plate is connected to the bottom plate, holes are formed in the plastic plate corresponding to the culture holes, tangible screens are respectively connected in the holes, the plastic plate is clamped with the bottom plate through clamping blocks, and clamping grooves are formed in the bottom plate corresponding to the clamping blocks. The liquid exchange device has the advantages that the liquid exchange is more complete, the liquid exchange of all culture holes can be completed by one-time operation, time and labor are saved, and the cultured cell cartilage balls can not be sucked out by misoperation.
Description
Technical Field
The utility model relates to the technical field of cell culture containers, in particular to a cell culture plate with a filtering device for stem cell chondrogenic differentiation.
Background
In most stem cell experimental studies, induction of differentiation into bone cells, fat cells, cartilage cells has become the gold standard for identifying their differentiation potential. The process of chondrogenic differentiation of mesenchymal stem cells includes that mesenchymal stem cells firstly withdraw the protuberance and aggregate into a cluster, cells in the middle of the cluster are divided and differentiated into a large round cell, and the large round cell is a chondroblast, and the chondroblast generates matrix and fiber (mainly type II collagen), and when the amount of the matrix is increased to a certain extent, the chondroblast is separated in a trap and differentiated into mature chondrocyte. The identification of in vitro induced chondrogenic differentiation is mainly carried out by morphological observation, aliskirin blue staining and detection of cartilage-forming markers, namely type II collagen, wherein the most commonly used aliskirin blue staining mainly detects acidic mucopolysaccharides in chondroblasts.
Cell culture plates are laboratory equipment for cell culture and are generally made of transparent plastic or glass materials, and comprise a plate and a cover, wherein the plate is provided with a plurality of small holes for accommodating cell growth. Cell culture plates are of many kinds, and there are 6, 12, 24, 48, 96, 384 and the like depending on the number of wells. The round bottom can be divided into a U-shaped bottom and a V-shaped bottom according to the shape of the bottom.
When the mesenchymal stem cells are subjected to cartilage forming differentiation culture, a 96-hole U-shaped culture plate is generally used, a certain concentration of mesenchymal stem cell suspension is added into a culture medium, after the cells are aggregated, the differentiation culture medium is replaced every 3 days, and then the differentiation culture medium is replaced every 3 days until the cartilage balls are fully differentiated. The cartilage balls were then stained for identification using the reagents in the aliskirin blue staining kit.
The cell culture plate structure of present cell culture plate structure generally its structure is as generally disclosed in patent application number "CN202321828638.5", including device main part and apron, the side fixed block is installed at the rear of device main part, the apron is installed in the top of device main part, the internally mounted of apron has the fixed screw, the experiment book is installed to the below of fixed clamp plate, the string bag is installed in the left side of draw-in groove, the surface mounting of dead lever has the stopper, the culture lid is installed to the top of culture box, and the top of culture lid is provided with the handle to the surface mounting of culture lid has the magnifying glass, device main part bottommost part is provided with the bottom plate, the inside of ventilation mouth is provided with the filter vent, the surface mounting of device main part has the bleeder vent. However, the utility model is used as a cell culture plate in the chondrogenic differentiation process of stem cells to perform a mesenchymal stem cell chondrogenic differentiation experiment, and the liquid exchange operation is very difficult when the suspension cartilage balls are cultured.
When the cells are changed, the old culture medium in the culture hole is sucked out, and then fresh culture medium is added. The existing liquid changing method only can be used for carefully sucking the liquid in the 96-well plate by using a liquid-transfering gun with a proper suction head.
The volume of cartilage balls formed by the mesenchymal stem cells at the beginning of differentiation is very small, the diameter is only less than 1mm, the color is white, and the cartilage balls are not easy to distinguish on a transparent culture plate. Because the culture hole is open, and the caliber of the used pipette tip is larger than the diameter of the cartilage ball, the cartilage ball is very easy to be sucked out and discarded together with the old culture medium under the action of suction force, and the experiment is failed. Moreover, since the cartilage balls are suspended in the medium, touching of the pipette tip may destroy newly formed cartilage balls, affecting the growth state thereof. In the dyeing identification step, the fixing solution, the dyeing solution and the cleaning solution are added and discarded for a plurality of times. Also, the cartilage balls are easily lost or destroyed. Therefore, the operator must take great care in changing the liquid.
Therefore, in order to prevent the suspended cartilage balls from being sucked, only the upper liquid is sucked as much as possible, and a part of liquid needs to be remained in the pore plate, so that the liquid change is incomplete, and the cell differentiation process and the staining result are influenced to a certain extent.
Moreover, the operation of each hole consumes a long time, when the number of holes used for culturing chondrocytes in the experiment is large, the workload of an experimenter can be greatly increased, the cells are in an exposed environment for a long time, the risk of cell pollution is easily increased, and the growth state of the cells is also influenced by the long-time operation.
Accordingly, the present utility model proposes a cell culture plate with a filter device for chondrogenic differentiation of stem cells, for solving the above-mentioned problems.
Disclosure of utility model
The utility model aims to provide a cell culture plate with a filtering device for stem cell chondrogenic differentiation, which is used for solving the problems that in the prior art, the operation of liquid exchange is difficult, newly formed cartilage balls are easily damaged by touching a suction head of a liquid-transfering gun, or the liquid exchange is incomplete, and the growth state of cells is influenced.
The technical scheme adopted for solving the technical problems is as follows:
A cell culture plate with a filtering device for chondrogenic differentiation of stem cells comprises a bottom plate and a plate cover detachably connected to the bottom plate, wherein a plurality of culture holes are formed in the bottom plate, a plastic plate is connected to the bottom plate, holes are formed in the plastic plate corresponding to the culture holes, tangible screens are respectively connected in the holes, the plastic plate is clamped with the bottom plate through clamping blocks, and clamping grooves are formed in the bottom plate corresponding to the clamping blocks.
By adopting the technical scheme, the culture medium can be completely abandoned as much as possible, the liquid exchange is more complete, and the liquid exchange of all culture holes can be completed by one-time operation, so that the time and the labor are saved.
Further, a plurality of anti-skid grooves are uniformly formed in the bottom of the clamping block.
By adopting the technical scheme, the plastic plate can be conveniently detached.
Further, the plastic plate comprises an upper plate and a lower plate, the upper plate is arranged above the lower plate, a shape screen is clamped between the upper plate and the lower plate, and two clamping blocks are symmetrically connected to two sides of the upper plate.
Through adopting above-mentioned technical scheme, can change the screen cloth, the screen cloth of different cell culture correspondence different mesh numbers, both can hold back the cartilage ball that experimental culture obtained, avoided the screen cloth too closely to lead to the difficult outflow of liquid again.
Further, a plurality of fixing nails are connected to the positions, corresponding to the shape screen mesh, of the bottoms of the upper plates, and nail grooves are correspondingly formed in the lower plates.
Through adopting above-mentioned technical scheme, can fix the screen cloth, guarantee the fixity of screen cloth, accurate entrapment experiment cultivates the cartilage ball that obtains.
Further, the bottom of lower plate is connected with the sealing strip.
By adopting the technical scheme, the compactness of the lower plate and the bottom plate can be ensured.
Further, a water suction groove is formed in the top of the upper plate.
By adopting the technical scheme, mixed flow of culture medium, staining solution and the like between holes can be avoided when the bottom plate is reversely buckled to discard the solution, and cross contamination is avoided.
In summary, compared with the prior art, the utility model has the beneficial effects that:
1. The utility model has simple operation of changing liquid, and can easily discard the liquid when intercepting the cartilage ball by only installing the screen and reversely buckling the bottom plate on the water absorbing paper. The cultured cell cartilage ball is not easy to be sucked out by misoperation, and the phenomenon that the suction head of the liquid-transfering gun touches the cartilage ball to damage the structure of the cartilage ball and influence the differentiation effect of the finished cartilage is avoided. The pipette is not required to be used for sucking liquid and discarding liquid, and the suction head is frequently replaced, so that liquid replacement of a plurality of culture holes can be realized at one time, and time and labor are saved.
2. The mesh number and the pore diameter of the screen mesh can be changed according to the size of the cultured cell mass, and the target cell mass can be trapped by the proper pore diameter of the screen mesh without increasing the flowing pressure of the liquid. And a baffle plate is designed between the screen holes, so that when the liquid is filtered by the back-off, the liquid in each hole is not mixed, and cross contamination is caused in culture.
Drawings
FIG. 1 is a schematic diagram of an explosive structure of the present utility model;
FIG. 2 is a schematic view of the working state of the present utility model;
FIG. 3 is a top view of FIG. 2;
FIG. 4 is a schematic view in section A-A of FIG. 3;
FIG. 5 is an enlarged partial schematic view of portion A of FIG. 4;
In the figure, 1, a bottom plate; 2, a plate cover, 3, a culture hole, 4, a hole site, 5, a screen, 6, a clamping block, 7, a clamping groove, 8, an anti-slip groove, 9, an upper plate, 10, a lower plate, 11, a fixing nail, 12, a nail groove, 13, a sealing strip, 14 and a water absorbing groove.
Detailed Description
For the purpose of making the objects, technical solutions and advantages of the embodiments of the present utility model more apparent, the technical solutions of the embodiments of the present utility model will be clearly and completely described below with reference to the accompanying drawings in the embodiments of the present utility model, and it is apparent that the described embodiments are some embodiments of the present utility model, but not all embodiments of the present utility model. All other embodiments, which can be made by those skilled in the art based on the embodiments of the utility model without making any inventive effort, are intended to be within the scope of the utility model.
In the present application, the azimuth or positional relationship indicated by the terms "upper", "inner", "outer", "middle", and the like are based on the azimuth or positional relationship shown in the drawings. These terms are only used to better describe the present application and its embodiments and are not intended to limit the scope of the indicated devices, elements or components to the particular orientations or to configure and operate in the particular orientations.
As shown in figures 1 and 2, a cell bottom plate 1 with a filtering device for chondrogenic differentiation of stem cells comprises a bottom plate 1 and a plate cover 2 detachably connected to the bottom plate 1, wherein a plurality of culture holes 3 are formed in the bottom plate 1, a plastic plate is connected to the bottom plate 1, holes 4 are formed in the plastic plate corresponding to the culture holes 3, and a screen 5 is respectively connected in the holes 4, so that each culture hole 3 can be detached. The plastic plate is clamped with the bottom plate 1 through the clamping blocks 6, and the bottom plate 1 is provided with clamping grooves 7 corresponding to the clamping blocks 6. After the cell culture is completed, the plate cover 2 is opened, the screen 5 in the independent sterile package is taken out, the screen 5 is installed on the bottom plate 1 through the plastic plate in a clamping manner, the bottom plate 1 can be further reversely buckled on sterile absorbent paper, the culture medium which needs to be abandoned is filtered, the cartilage balls are trapped in the screen 5, the culture medium can be completely abandoned as much as possible, the liquid exchange is more complete, the liquid exchange of all culture holes 3 can be completed through one-time operation, and the time and labor are saved. Wherein a plurality of anti-skid grooves 8 are uniformly formed in the bottom of the clamping block 6. The plastic plate is convenient to detach.
As shown in fig. 3 and 4, the plastic plate comprises an upper plate 9 and a lower plate 10, the upper plate 9 is arranged above the lower plate 10, a screen 5 is clamped between the upper plate 9 and the lower plate 10, and two clamping blocks 6 are symmetrically connected to two sides of the upper plate 9. Furthermore, the screen 5 can be replaced, and different cell cultures correspond to the screen 5 with different meshes, so that not only can cartilage balls obtained by experimental culture be retained, but also liquid is prevented from flowing out due to too dense screen 5.
As shown in fig. 4, the top of the upper plate 9 is provided with a water suction groove 14. Further, the sterile absorbent paper is placed in the absorbent tank 14, and then the bottom plate 1 with the screen 5 is inverted on the sterile absorbent paper, so that mixed flow of culture medium, staining solution and the like between holes can be avoided when the bottom plate 1 is reversely buckled to discard the solution, and cross contamination is avoided.
As shown in fig. 5, a plurality of fixing nails 11 are connected to the bottom of the upper plate 9 corresponding to the screen 5, and nail grooves 12 are correspondingly formed in the lower plate 10. Can fix screen cloth 5, guarantee screen cloth 5's fixity, accurate entrapment experiment cultivates the cartilage ball that obtains. The bottom of the lower plate 10 is connected with a sealing strip 13. The compactness of the lower plate 10 and the bottom plate 1 is ensured, and mixed flow of culture medium, staining solution and the like among all holes is avoided when the bottom plate 1 is reversely buckled to discard the solution.
The working process of the utility model is as follows:
The mesenchymal stem cells were first suspended in the chondrogenic differentiation medium in the culture well 3, and covered with the plate cover 2. The bottom plate 1 was placed in an incubator for culture when aggregation of cells was observed. The screen 5 in an independent sterile package is opened, the screen 5 is fixed through the upper plate 9 and the lower plate 10, the screen 5 is clamped and installed on the bottom plate 1 through the clamping blocks, then the sterile water absorbing paper is placed in the water absorbing groove 14, and then the bottom plate 1 with the screen 5 is inverted on the sterile water absorbing paper, so that the culture medium in each culture hole 3 flows out. After the complete outflow of the medium, the bottom plate 1 is placed upright and the cartilage balls are then trapped in the culture holes 3 by the filter screen 5. The screen 5 was removed, and the chondrogenic differentiation medium was continuously added to each culture well 3, covered with the plate cover 2, and placed in an incubator for culturing for 14 days, during which the chondrogenic differentiation medium was replaced in the same manner every 3 days. After the culture is completed, the cartilage ball is subjected to the next work.
Claims (6)
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202420664189.3U CN222099996U (en) | 2024-04-02 | 2024-04-02 | A cell culture plate with a filter device for chondrogenic differentiation of stem cells |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202420664189.3U CN222099996U (en) | 2024-04-02 | 2024-04-02 | A cell culture plate with a filter device for chondrogenic differentiation of stem cells |
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| Publication Number | Publication Date |
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| CN222099996U true CN222099996U (en) | 2024-12-03 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN202420664189.3U Active CN222099996U (en) | 2024-04-02 | 2024-04-02 | A cell culture plate with a filter device for chondrogenic differentiation of stem cells |
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| CN (1) | CN222099996U (en) |
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- 2024-04-02 CN CN202420664189.3U patent/CN222099996U/en active Active
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