CN111154649A - Seal for manufacturing multi-concave gel sheet and using method - Google Patents
Seal for manufacturing multi-concave gel sheet and using method Download PDFInfo
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- CN111154649A CN111154649A CN202010127336.XA CN202010127336A CN111154649A CN 111154649 A CN111154649 A CN 111154649A CN 202010127336 A CN202010127336 A CN 202010127336A CN 111154649 A CN111154649 A CN 111154649A
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- 238000004519 manufacturing process Methods 0.000 title claims abstract description 20
- 238000000034 method Methods 0.000 title claims abstract description 13
- 238000004113 cell culture Methods 0.000 claims abstract description 36
- 238000004891 communication Methods 0.000 claims description 3
- 238000001816 cooling Methods 0.000 claims description 3
- 230000008901 benefit Effects 0.000 abstract description 4
- 230000004907 flux Effects 0.000 abstract description 4
- 239000000499 gel Substances 0.000 description 39
- 239000000243 solution Substances 0.000 description 19
- 210000004027 cell Anatomy 0.000 description 15
- 238000001727 in vivo Methods 0.000 description 3
- 206010028980 Neoplasm Diseases 0.000 description 2
- 239000000090 biomarker Substances 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- 238000012136 culture method Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 239000000463 material Substances 0.000 description 2
- 239000008188 pellet Substances 0.000 description 2
- 229920000936 Agarose Polymers 0.000 description 1
- 102000009027 Albumins Human genes 0.000 description 1
- 108010088751 Albumins Proteins 0.000 description 1
- 206010019851 Hepatotoxicity Diseases 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- XSQUKJJJFZCRTK-UHFFFAOYSA-N Urea Chemical compound NC(N)=O XSQUKJJJFZCRTK-UHFFFAOYSA-N 0.000 description 1
- 238000004115 adherent culture Methods 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 239000004202 carbamide Substances 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 210000003494 hepatocyte Anatomy 0.000 description 1
- 230000007686 hepatotoxicity Effects 0.000 description 1
- 231100000304 hepatotoxicity Toxicity 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 210000005229 liver cell Anatomy 0.000 description 1
- 230000003908 liver function Effects 0.000 description 1
- 108020004999 messenger RNA Proteins 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 238000002331 protein detection Methods 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 230000003248 secreting effect Effects 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 238000010186 staining Methods 0.000 description 1
- 229910001220 stainless steel Inorganic materials 0.000 description 1
- 239000010935 stainless steel Substances 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
- C12M25/00—Means for supporting, enclosing or fixing the microorganisms, e.g. immunocoatings
- C12M25/14—Scaffolds; Matrices
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- Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Sustainable Development (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Immunology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention relates to a seal for manufacturing a multi-concave gel sheet and a using method thereof, the seal comprises a seal die and a handle detachably connected with the seal die, the seal die is provided with an upper surface and a lower surface which are oppositely arranged, and the lower surface is provided with a plurality of bulges; and a pouring gate is further arranged on the die and communicated with the lower surface. Through be provided with a plurality of evenly distributed's arch on the lower surface, and be provided with on the die with the runner of lower surface intercommunication, when placing the die in the cell culture apparatus, gel solution pours into and flows into to the space of lower surface and cell culture apparatus between in order to form the gel piece through this runner, and fashioned gel piece has a plurality of concave surface in order to obtain a plurality of cell balls, and its simple structure and cost are lower, still has the advantage of high flux simultaneously.
Description
Technical Field
The invention relates to a seal for a multi-concave-surface gel sheet and a using method thereof.
Background
The conventional cell culture is an adherent two-dimensional culture mode, but because the communication among cells is very little, the cell group cannot have the structure and physiological and biochemical characteristics similar to those in vivo. For example, liver cells lose the hepatic ductuli and function of secreting albumin/urea several days later in adherent culture, or cancer cells lack biomarkers in vivo in two-dimensional culture. In contrast, the three-dimensional culture of the cells can effectively improve and simulate the in vivo environment, for example, the three-dimensional culture of the hepatocytes has the three-dimensional structure and the liver function of a natural liver, and more accurate hepatotoxicity data is obtained; the cancer cells can reproduce some biomarkers and certain metastasis and infiltration functions under three-dimensional culture. Therefore, the importance of three-dimensional cell culture is self evident.
However, the current three-dimensional culture method is too complex and expensive (such as a scaffold method), so that the three-dimensional culture method cannot be widely accepted by ordinary researchers. Or lack of high throughput features, such as the use of agarose or ultra-low adsorption plates in 96-well plates, where only one cell three-dimensional sphere can be formed in one well; since each cell pellet has only about 1000 cells, the criteria for protein detection, qPCR quantification of mRNA, cell pellet staining, and the lowest cell number of most kits cannot be met.
Disclosure of Invention
The invention aims to provide a seal for manufacturing a multi-concave-surface gel sheet and a using method thereof, which are used in a cell culture tool so that the cell culture tool has the advantage of high flux, and is low in cost and simple in structure.
In order to achieve the purpose, the invention provides the following technical scheme: a seal for manufacturing a multi-concave gel sheet comprises a seal die and a handle detachably connected with the seal die, wherein the seal die is provided with an upper surface and a lower surface which are oppositely arranged, and a plurality of bulges are arranged on the lower surface; and a pouring gate is further arranged on the die and communicated with the lower surface.
Further, the protrusion comprises a first bump, and the shape of the cross section of the first bump is semicircular, conical or rectangular.
Further, the bump further includes a second bump for connecting the first bump and the die, and a cross section of the second bump is rectangular.
Furthermore, a groove formed by inwards recessing the upper surface is further formed in the die, and the pouring gate is communicated with the groove.
Further, the handle is provided with a clamping block clamped with the groove.
Further, the cross-section of the runner is tapered in shape.
Further, the diameter range of the protrusions is 0.4-5mm, and the depth of the protrusions is 0.2-2.5 mm.
The invention also provides a method for manufacturing the multi-concave surface gel sheet, which adopts the seal for manufacturing the multi-concave surface gel sheet, and comprises the following steps:
preparing a gel solution;
placing the stamp into a cell culture tool;
flowing a thermal gel solution into a gap between the lower surface of the stamp and the cell culture tool through the runner until the thermal gel solution fills the gap;
cooling the hot gel solution until the hot gel solution is completely solidified;
inserting a handle to enable the handle to be clamped with the die, enabling the die to be separated from the solidified multi-concave gel sheet, and removing the die out of the cell culture tool.
Further, the method further comprises:
preheating the stamp before placing the stamp in the cell culture tool.
Further, the cell culture tool is any one of a 24-well plate, a 12-well plate, a 6-well plate, a 35mm cell culture dish, and a 60mm cell culture dish.
The invention has the beneficial effects that: through be provided with a plurality of evenly distributed's arch on the lower surface, and be provided with on the die with the runner of lower surface intercommunication, when placing the die in the cell culture apparatus, gel solution pours into and flows into to the space of lower surface and cell culture apparatus between in order to form the gel piece through this runner, and fashioned gel piece has a plurality of concave surface in order to obtain a plurality of cell balls, and its simple structure and cost are lower, still has the advantage of high flux simultaneously.
The foregoing description is only an overview of the technical solutions of the present invention, and in order to make the technical solutions of the present invention more clearly understood and to implement them in accordance with the contents of the description, the following detailed description is given with reference to the preferred embodiments of the present invention and the accompanying drawings.
Drawings
Fig. 1 is a schematic structural diagram of a stamp for manufacturing a multi-concave gel sheet according to the present invention.
Fig. 2 is a schematic view of the structure of the stamp.
Fig. 3 is another schematic view of the stamp.
Fig. 4 is a schematic cross-sectional view of a stamp.
Fig. 5 is a schematic view of the structure of the protrusion.
Fig. 6 is a schematic structural view of the handle.
Detailed Description
The following detailed description of embodiments of the present invention is provided in connection with the accompanying drawings and examples. The following examples are intended to illustrate the invention but are not intended to limit the scope of the invention.
Referring to fig. 1 to 6, a stamp for manufacturing a multi-concave gel sheet according to a preferred embodiment of the present invention is used in a cell culture tool, and in this embodiment, the cell culture tool may be any one of a 24-well plate, a 12-well plate, a 6-well plate, a 35mm cell culture dish, and a 60mm cell culture dish. However, in other embodiments, the cell culture tool may be other, and is not particularly limited herein, depending on the actual situation. If the seal for manufacturing the multi-concave-surface gel sheet is applied to cell culture tools of different models, the size of the seal is correspondingly changed, and the embodiment does not specifically limit the size of the seal for manufacturing the multi-concave-surface gel sheet and is determined according to actual conditions.
The seal for manufacturing the multi-concave gel sheet comprises a stamp 1 and a handle 2 detachably connected with the stamp 1, wherein the stamp 1 and the handle 2 are made of stainless steel materials, and the seal is high in hardness, wear-resistant, high-temperature-resistant and sterilizable. Wherein the smoothness of the stamp 1 is up to the nanometer level in order to improve the smoothness of the molded gel sheet. Indeed, in other embodiments, the impression 1 and the handle 2 may be made of other materials, which are determined by the actual situation, and are not specifically limited herein.
The lower surface 12 is provided with a plurality of protrusions 15, and the plurality of protrusions 15 are uniformly distributed on the lower surface 12. Indeed, in other embodiments, the plurality of protrusions 15 may be distributed unevenly, and are not limited herein, depending on the actual situation. In this embodiment, the number of the protrusions 15 is 566. Indeed, in other embodiments, the number of the protrusions 15 may be other, and is not limited in particular, according to the actual situation.
The diameter range of the protrusion 15 is 0.4-5mm, and the depth of the protrusion 15 is 0.2-2.5mm, but in other embodiments, the diameter range and the depth range of the protrusion 15 may be other, and are not limited herein according to the actual situation. The protrusion 15 includes a first protrusion 151, and the shape of the cross section of the first protrusion 151 is a semicircle, a cone, a rectangle, or the like, and is not particularly limited herein, and the shape of the cross section of the first protrusion 151 is preferably a semicircle, which is better to form a cell sphere. The bump 15 further includes a second bump 152 for connecting the first bump 151 and the die 1, and the cross section of the second bump 152 is rectangular, and the like, which is not limited herein. The second bump 152 is disposed according to actual requirements, that is, the second bump 152 may or may not be disposed.
The die 1 is further provided with a pouring channel 14, and the pouring channel 14 is communicated with the lower surface 12. In order to facilitate the injection of the gel solution and to reduce the difficulty of the manufacturing process of the stamp 1, the runner 14 is disposed in communication with the recess 13 in this embodiment. And injecting the gel solution into the groove 13, and then flowing to the lower surface 12 of the stamp 1 through the pouring channel 14 until the gap between the lower surface 12 and the pore plate is fully paved, so that the method is convenient and quick. Indeed, in other embodiments, the runner 14 may also communicate with the side wall of the stamp 1, but this is less convenient.
After the gel sheet is formed in the cell culture tool, the gel sheet has a plurality of concave surfaces matching with the projections 15, and after the cell suspension is added into the cell culture tool, the cells will slowly gather in each concave surface of the multi-concave surface gel sheet, and the cell can be formed into a ball after three days.
The invention also provides a method for manufacturing the multi-concave surface gel sheet, which adopts the seal for manufacturing the multi-concave surface gel sheet, and comprises the following steps:
preparing a gel solution; the gel solution is a hydrogel solution or others, and is not particularly limited herein, depending on the actual situation.
Placing the stamp 1 into a cell culture tool;
flowing a thermal gel solution through the runner 14 into the gap between the lower surface 12 of the stamp 1 and the cell culture tool until the gap is filled with the thermal gel solution;
cooling the hot gel solution until the hot gel solution is completely solidified;
inserting a handle 2 to engage the handle 2 with the stamp 1, separating the stamp 1 from the solidified multi-concave gel sheet, and removing the stamp 1 from the cell culture tool.
Wherein, the method of the invention also comprises:
before placing the stamp 1 in the cell culture tool, the stamp 1 is preheated. The purpose of preheating is to prevent the added gel solution from being cooled and shaped too quickly, thereby affecting the final shaped product.
In summary, the following steps: through being provided with a plurality of evenly distributed's arch on lower surface 12, and be provided with runner 14 with lower surface 12 intercommunication on the impression 1, when placing impression 1 in the cell culture apparatus, the gel solution pours into and flows into through this runner 14 and to form the gel piece between the space of lower surface and cell culture apparatus, and the fashioned gel piece has a plurality of concave surfaces in order to obtain a plurality of cell balls, and its simple structure and cost are lower, still has the advantage of high flux simultaneously.
The technical features of the embodiments described above may be arbitrarily combined, and for the sake of brevity, all possible combinations of the technical features in the embodiments described above are not described, but should be considered as being within the scope of the present specification as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is more specific and detailed, but not construed as limiting the scope of the invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present patent shall be subject to the appended claims.
Claims (10)
1. A seal for manufacturing a multi-concave gel sheet is characterized by comprising a seal die and a handle detachably connected with the seal die, wherein the seal die is provided with an upper surface and a lower surface which are oppositely arranged, and a plurality of bulges are arranged on the lower surface; and a pouring gate is further arranged on the die and communicated with the lower surface.
2. The stamp for making a multi-concave gel sheet according to claim 1, wherein the protrusions comprise first protrusions, and the cross-section of the first protrusions has a shape of a semicircle, a cone, or a rectangle.
3. The stamp for making multi-concave gel sheets according to claim 2, wherein the protrusions further comprise second protrusions for connecting the first protrusions and the stamp, the second protrusions having a rectangular cross-sectional shape.
4. The stamp for making multiple concave gel sheets according to claim 1, wherein the stamp further has a recess formed by recessing the upper surface inward, and the runner is in communication with the recess.
5. The stamp for making multiple concave gel sheets according to claim 4, wherein said handle has a latch for engaging with said recess.
6. The stamp for making multiple concave gel sheets according to claim 1, wherein the cross-section of the runner is tapered in shape.
7. The stamp for making multi-concave gel sheets according to claim 1, wherein the diameter of the protrusions ranges from 0.4 to 5mm, and the depth of the protrusions ranges from 0.2 to 2.5 mm.
8. A method for manufacturing a multi-concave gel sheet, wherein the stamp for manufacturing a multi-concave gel sheet according to any one of claims 1 to 5 is used, the method comprising:
preparing a gel solution;
placing the stamp into a cell culture tool;
flowing a thermal gel solution into a gap between the lower surface of the stamp and the cell culture tool through the runner until the thermal gel solution fills the gap;
cooling the hot gel solution until the hot gel solution is completely solidified;
inserting a handle to enable the handle to be clamped with the die, enabling the die to be separated from the solidified multi-concave gel sheet, and removing the die out of the cell culture tool.
9. The method of making a multi-concave gel sheet of claim 8, further comprising:
preheating the stamp before placing the stamp in the cell culture tool.
10. The method for producing a multi-concave gel sheet according to claim 8, wherein the cell culture tool is any one of a 24-well plate, a 12-well plate, a 6-well plate, a 35mm cell culture dish, and a 60mm cell culture dish.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010127336.XA CN111154649A (en) | 2020-02-28 | 2020-02-28 | Seal for manufacturing multi-concave gel sheet and using method |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202010127336.XA CN111154649A (en) | 2020-02-28 | 2020-02-28 | Seal for manufacturing multi-concave gel sheet and using method |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN111154649A true CN111154649A (en) | 2020-05-15 |
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|---|---|---|---|
| CN202010127336.XA Pending CN111154649A (en) | 2020-02-28 | 2020-02-28 | Seal for manufacturing multi-concave gel sheet and using method |
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Application publication date: 20200515 |
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