CN219752329U - Stem cell culture device for rapid detection - Google Patents
Stem cell culture device for rapid detection Download PDFInfo
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- CN219752329U CN219752329U CN202320457354.3U CN202320457354U CN219752329U CN 219752329 U CN219752329 U CN 219752329U CN 202320457354 U CN202320457354 U CN 202320457354U CN 219752329 U CN219752329 U CN 219752329U
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- 210000000130 stem cell Anatomy 0.000 title claims abstract description 41
- 238000001514 detection method Methods 0.000 title claims abstract description 25
- 238000004113 cell culture Methods 0.000 title claims abstract description 21
- 238000000605 extraction Methods 0.000 claims abstract description 21
- 239000012528 membrane Substances 0.000 claims abstract description 12
- 238000009423 ventilation Methods 0.000 claims abstract description 8
- 238000003556 assay Methods 0.000 claims 5
- 239000000243 solution Substances 0.000 description 29
- 238000000034 method Methods 0.000 description 9
- 239000003153 chemical reaction reagent Substances 0.000 description 8
- 108090000790 Enzymes Proteins 0.000 description 6
- 102000004190 Enzymes Human genes 0.000 description 6
- 229940088598 enzyme Drugs 0.000 description 6
- 238000006243 chemical reaction Methods 0.000 description 5
- 230000000593 degrading effect Effects 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 230000008569 process Effects 0.000 description 5
- 238000012258 culturing Methods 0.000 description 4
- 239000003814 drug Substances 0.000 description 4
- 238000004020 luminiscence type Methods 0.000 description 4
- 230000035945 sensitivity Effects 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 230000003287 optical effect Effects 0.000 description 3
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 2
- 239000005089 Luciferase Substances 0.000 description 2
- 239000012496 blank sample Substances 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 230000001172 regenerating effect Effects 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 description 2
- IGXWBGJHJZYPQS-SSDOTTSWSA-N D-Luciferin Chemical compound OC(=O)[C@H]1CSC(C=2SC3=CC=C(O)C=C3N=2)=N1 IGXWBGJHJZYPQS-SSDOTTSWSA-N 0.000 description 1
- CYCGRDQQIOGCKX-UHFFFAOYSA-N Dehydro-luciferin Natural products OC(=O)C1=CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 CYCGRDQQIOGCKX-UHFFFAOYSA-N 0.000 description 1
- BJGNCJDXODQBOB-UHFFFAOYSA-N Fivefly Luciferin Natural products OC(=O)C1CSC(C=2SC3=CC(O)=CC=C3N=2)=N1 BJGNCJDXODQBOB-UHFFFAOYSA-N 0.000 description 1
- 108060001084 Luciferase Proteins 0.000 description 1
- DDWFXDSYGUXRAY-UHFFFAOYSA-N Luciferin Natural products CCc1c(C)c(CC2NC(=O)C(=C2C=C)C)[nH]c1Cc3[nH]c4C(=C5/NC(CC(=O)O)C(C)C5CC(=O)O)CC(=O)c4c3C DDWFXDSYGUXRAY-UHFFFAOYSA-N 0.000 description 1
- 102000004317 Lyases Human genes 0.000 description 1
- 108090000856 Lyases Proteins 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 108091005804 Peptidases Proteins 0.000 description 1
- 239000004365 Protease Substances 0.000 description 1
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- 239000007864 aqueous solution Substances 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 229960000686 benzalkonium chloride Drugs 0.000 description 1
- CADWTSSKOVRVJC-UHFFFAOYSA-N benzyl(dimethyl)azanium;chloride Chemical compound [Cl-].C[NH+](C)CC1=CC=CC=C1 CADWTSSKOVRVJC-UHFFFAOYSA-N 0.000 description 1
- 238000005415 bioluminescence Methods 0.000 description 1
- 230000029918 bioluminescence Effects 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000002894 multi-fate stem cell Anatomy 0.000 description 1
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- 238000011160 research Methods 0.000 description 1
- 239000000523 sample Substances 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
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- 238000012360 testing method Methods 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- LENZDBCJOHFCAS-UHFFFAOYSA-N tris Chemical compound OCC(N)(CO)CO LENZDBCJOHFCAS-UHFFFAOYSA-N 0.000 description 1
Landscapes
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The utility model belongs to the technical field of cell culture, and discloses a stem cell culture device for rapid detection, which comprises a cylindrical base with an opening at the upper part and a closed lower part, wherein an extraction layer and a culture layer are sequentially spliced and connected at the upper part of the base upwards, the upper parts of the extraction layer and the culture layer are both opened, and the lower parts of the extraction layer and the culture layer are both sealed by a semipermeable membrane; the side wall of the culture layer is provided with a plurality of ventilation holes which are closed by a semipermeable membrane; the upper part of the culture layer is connected with a top cover in a plug-in way. The utility model can culture and detect the stem cells, and meet the requirement of rapid detection of the stem cells.
Description
Technical Field
The utility model belongs to the technical field of cell culture, and particularly relates to a stem cell culture device for rapid detection.
Background
Stem cells are multipotent cells with self-replication capacity, are insufficiently differentiated and immature cells, have potential functions of regenerating various tissues and organs and human bodies, and are called as universal cells in the medical field; under certain conditions, it can differentiate into a variety of functional cells. Therefore, the method has important significance in basic research and transformation medicine application, and has wide application prospect in the fields of regenerative medicine, disease models, medicine screening, accurate medicine and the like.
However, in the stem cell culture process, the culture device is too single, and is difficult to obtain rapid treatment in the process of collection and observation, so that the detection process is complicated and long, and therefore, a culture device capable of being used for rapid detection needs to be designed.
Disclosure of Invention
The technical problems to be solved by the utility model are as follows: the stem cell culture device for rapid detection overcomes the defects of the prior art, can culture and detect the stem cells, and meets the requirement of rapid detection of the stem cells.
In order to solve the technical problems, the technical scheme of the utility model is as follows:
a stem cell culture device for rapid detection comprises a cylindrical base with an upper opening and a lower closed, wherein an extraction layer and a culture layer are sequentially spliced and connected on the upper part of the base upwards, the upper parts of the extraction layer and the culture layer are both opened, and the lower parts of the extraction layer and the culture layer are both sealed by a semipermeable membrane; the side wall of the culture layer is provided with a plurality of ventilation holes which are closed by a semipermeable membrane; the upper part of the culture layer is connected with a top cover in a plug-in way.
Preferably, the closed end of the lower part of the base is transparent. Fluorescent agent is arranged in the base, and the fluorescent agent can be directly detected by being placed on a detecting instrument provided with a light detector after reaction.
Preferably, the base, the extraction layer, the culture layer and the top cover are arranged coaxially and with the same diameter.
Preferably, 4 to 8 ventilation holes are formed and are symmetrically and uniformly formed on the side wall of the culture layer respectively. Ensuring gas exchange.
Preferably, an extraction reagent is arranged in the extraction layer. The stem cell internal substance can be extracted. The subsequent reaction and detection are convenient.
Preferably, a culture solution is arranged in the culture layer.
Preferably, the stem cell culture device is further provided with a scalpel and a syringe.
Preferably, the stem cell culture apparatus performs stem cell detection by a light detector disposed and a computer that calculates the amount of luminescence from a detection signal of the light detector and determines the growth state of stem cells based on the temporal change of the amount of luminescence.
Due to the adoption of the technical scheme, the utility model has the beneficial effects that:
1. the method for detecting the stem cell ATP by bioluminescence through the luciferin-luciferase reaction has the characteristic of high sensitivity, and overcomes the defects of reduced luminous intensity and reduced sensitivity caused by the fact that the luminous reagent serving as the enzyme is inhibited by the ATP extraction reagent under the condition that the luminous reagent of the ATP extraction reagent is pre-mixed by separating the culture solution from the extracting solution and the luminous solution; by culturing stem cells and pre-treating the stem cells before detection, the detection time can be greatly shortened.
In a word, the utility model carries out the pretreatment before the culture and detection of stem cells, thus greatly shortening the detection time; the ATP method has the advantage of high sensitivity to stem cell detection, so that the ATP method has a wide application prospect.
Drawings
FIG. 1 is a schematic diagram of the structure of the present utility model;
in the figure, 1, a base; 2. extracting a layer; 3. a culture layer; 4. a semipermeable membrane; 5. ventilation holes; 6. and a top cover.
Detailed Description
The utility model is further illustrated in the following, in conjunction with the accompanying drawings and examples.
Embodiment one:
as shown in figure 1, the stem cell culture device for rapid detection comprises a cylindrical base 1 with an upper opening and a lower closed, wherein an extraction layer 2 and a culture layer 3 are sequentially spliced and connected on the upper part of the base 1, the upper parts of the extraction layer 2 and the culture layer 3 are respectively opened, and the lower parts of the extraction layer 2 and the culture layer 3 are respectively sealed by a semipermeable membrane 4; four ventilation holes 5 are formed in the side wall of the culture layer 3, and the ventilation holes 5 are closed by a semipermeable membrane (not marked); the upper part of the culture layer 3 is connected with a top cover 6 in a plug-in way.
In actual use, preparing a culture solution, an extracting solution and a luminescent solution, wherein the culture solution is a culture solution added with ATP degrading enzyme in a corresponding commercial stem cell culture solution, and the ATP degrading enzyme can eliminate ATP contained in the culture solution or free ATP contained in a sample so as to improve detection sensitivity; the extracting solution is ATP extracting solution, contains aqueous solution of benzalkonium chloride, trichloroacetic acid (TCA), tris buffer solution, ethanol, lyase with protease activity, lysozyme and other reagents, and can lyse stem cells and release ATP for reaction; the luminescent liquid is a reagent that can be mixed with ATP to emit light, for example, a reagent containing luciferase and luciferin.
The actual culturing process can be carried out in two ways, the first one: the culture layer 3 is inserted onto the base 1, then the culture solution added with the ATP degrading enzyme and the stem cells to be cultured are injected into the culture layer 3, the top cover 6 is covered, the culture solution is placed into the incubator for culturing, the ATP degrading enzyme is added into the culture solution through the injector (not shown) and the air holes 5, the air holes 5 are closed by the semipermeable membrane after the addition, the culture layer 3 and the base 1 are separated after the culture is completed for a certain time, the luminescent solution is injected into the base 1, the ATP extracting solution is injected into the culture layer 3, the extracting solution is completely mixed with the culture solution and the stem cells to be cultured, and after the stem cells are lysed, the mixture is poured into the base 1 for luminescence, and the detection can be carried out through the photodetector and the computer.
Second kind: injecting a luminescent liquid into the base 1, then plugging an extraction layer 2 on the luminescent liquid, injecting an ATP extraction liquid into the extraction layer 2, plugging a culture layer 3 on the extraction layer 2, injecting a culture liquid added with ATP degrading enzyme and stem cells to be cultured into the culture layer 3, covering a top cover 6, and placing the culture layer into an incubator for culturing; during the culture process, the culture solution can be supplemented into the culture layer 3 through the injector, after the culture is carried out for a certain time, the culture layer is taken out, the top cover 6 is opened, the semi-permeable membrane at the bottom of the culture layer 3 is scratched by the scalpel, the culture solution and the stem cells are fully mixed with the ATP extracting solution, after the stem cells are cracked, the semi-permeable membrane at the bottom of the extracting layer 2 is scratched by the scalpel, the reaction solution and the luminous solution are fully mixed, and then the culture device can be placed into a light detector connected with a computer for detection.
In the actual detection process, a photomultiplier, a CCD camera or a photodiode can be selected as a light detector, so that an optical signal is transmitted into a computer, the growth condition of stem cells can be determined by calculating the luminous quantity after the computer receives the detection signal, the luminous quantity is calculated by subtracting the optical signal value of a blank sample from the maximum value of the detected optical signal, and the blank sample is obtained by mixing a culture solution, an ATP extracting solution and a luminous solution; the relation between the amount of luminescence and the number of stem cells can be obtained by the test result of the standard substance.
It is to be understood that these examples are illustrative of the present utility model and are not intended to limit the scope of the present utility model. Furthermore, it should be understood that various changes and modifications can be made by one skilled in the art after reading the teachings of the present utility model, and such equivalents are intended to fall within the scope of the utility model as defined in the appended claims.
Claims (6)
1. A stem cell culture apparatus for rapid detection, characterized in that: the device comprises a cylindrical base with an opening at the upper part and a closed lower part, wherein the upper part of the base is connected with an extraction layer and a culture layer in an upward and sequential inserting manner, the upper parts of the extraction layer and the culture layer are both opened, and the lower parts are both sealed with a semi-permeable membrane; a plurality of ventilation holes are formed in the side wall of the culture layer, and the ventilation holes are closed by a semipermeable membrane; the upper part of the culture layer is connected with a top cover in a plug-in manner.
2. The rapid assay stem cell culture apparatus of claim 1, wherein: the closed end of the lower part of the base is transparent.
3. The rapid assay stem cell culture apparatus of claim 1, wherein: the base, the extraction layer, the culture layer and the top cover are coaxial and arranged with the same diameter.
4. The rapid assay stem cell culture apparatus of claim 1, wherein: 4 to 8 breather holes are formed.
5. The rapid assay stem cell culture apparatus of claim 1, wherein: the stem cell culture device is also provided with a scalpel and a syringe.
6. The rapid assay stem cell culture apparatus of claim 1, wherein: the lower part of the stem cell culture device can be provided with a light detector and a computer.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320457354.3U CN219752329U (en) | 2023-03-09 | 2023-03-09 | Stem cell culture device for rapid detection |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202320457354.3U CN219752329U (en) | 2023-03-09 | 2023-03-09 | Stem cell culture device for rapid detection |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN219752329U true CN219752329U (en) | 2023-09-26 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202320457354.3U Active CN219752329U (en) | 2023-03-09 | 2023-03-09 | Stem cell culture device for rapid detection |
Country Status (1)
| Country | Link |
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| CN (1) | CN219752329U (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117683634A (en) * | 2024-01-31 | 2024-03-12 | 山东格林医学科技有限公司 | Stem cell culture device and method for diabetes treatment |
-
2023
- 2023-03-09 CN CN202320457354.3U patent/CN219752329U/en active Active
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117683634A (en) * | 2024-01-31 | 2024-03-12 | 山东格林医学科技有限公司 | Stem cell culture device and method for diabetes treatment |
| CN117683634B (en) * | 2024-01-31 | 2024-05-07 | 山东格林医学科技有限公司 | Stem cell culture device and method for diabetes treatment |
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