CN109055218A - A kind of cell circle scar Healing Experiments device for six orifice plates - Google Patents
A kind of cell circle scar Healing Experiments device for six orifice plates Download PDFInfo
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- CN109055218A CN109055218A CN201811137312.1A CN201811137312A CN109055218A CN 109055218 A CN109055218 A CN 109055218A CN 201811137312 A CN201811137312 A CN 201811137312A CN 109055218 A CN109055218 A CN 109055218A
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- cell
- orifice plates
- scar
- scar healing
- criss
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- 231100000241 scar Toxicity 0.000 title claims abstract description 43
- 230000035876 healing Effects 0.000 title claims abstract description 32
- 238000002474 experimental method Methods 0.000 title claims abstract description 26
- 238000010276 construction Methods 0.000 claims abstract description 28
- 238000000034 method Methods 0.000 claims description 4
- 239000000463 material Substances 0.000 claims description 3
- 229910001220 stainless steel Inorganic materials 0.000 claims description 3
- 239000010935 stainless steel Substances 0.000 claims description 3
- 230000000007 visual effect Effects 0.000 abstract description 3
- 210000004027 cell Anatomy 0.000 description 41
- 230000005012 migration Effects 0.000 description 6
- 238000013508 migration Methods 0.000 description 6
- SHGAZHPCJJPHSC-YCNIQYBTSA-N all-trans-retinoic acid Chemical compound OC(=O)\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C SHGAZHPCJJPHSC-YCNIQYBTSA-N 0.000 description 5
- 229930002330 retinoic acid Natural products 0.000 description 5
- 230000012292 cell migration Effects 0.000 description 4
- 230000010261 cell growth Effects 0.000 description 3
- 230000000694 effects Effects 0.000 description 3
- 230000029663 wound healing Effects 0.000 description 3
- 238000003556 assay Methods 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 210000004185 liver Anatomy 0.000 description 2
- 238000012545 processing Methods 0.000 description 2
- 239000002356 single layer Substances 0.000 description 2
- 210000004881 tumor cell Anatomy 0.000 description 2
- 208000032544 Cicatrix Diseases 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 230000001464 adherent effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 239000013553 cell monolayer Substances 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 238000012258 culturing Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 230000003252 repetitive effect Effects 0.000 description 1
- 230000037387 scars Effects 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 238000010998 test method Methods 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
Classifications
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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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/12—Well or multiwell plates
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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)
- Clinical Laboratory Science (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
Abstract
The present invention provides a kind of cell scar Healing Experiments devices suitable for six orifice plates, including tablet (1), tablet bottom to be provided with criss-cross construction (2)；The cross end bottom of criss-cross construction is provided with cylindrical structure (3).The case where present invention provides a kind of round cell scar Healing Experiments device suitable for six orifice plates, facilitates the scar healing in observation whole visual field, avoids current cell scratch experiment that from can only observing the healing of scratch regional area.
Description
Technical field
The present invention relates to cell biologies, disclose a kind of cell scar Healing Experiments dress for preparing round scar
It sets.
Technical background
Cell wound healing assay is a kind of most common ill vitro test method for studying cell migration ability.Its base
Present principles are: when cell growth is fused to single layer state, one " scar " is artificially manufactured on single-layer culturing cell, edge
Cell, which can progress into white space, makes " scratch " to heal.By calculating the healing rate of cell scar, the migration energy of cell is assessed
Power simulates the process of internal cell migration to a certain extent.The factor of external change cell migration and/or growth can increase
Add or reduce wound healing rate, therefore, this experiment is usually used in studying various experiment conditions such as gene regulation and drug in cell
Effect in migration.This experimental implementation is simple, and expense is cheap.
Traditional cell wound healing assay method be with pipette tips gently on cell monolayer between scratch, scratch crosses via hole
Plate, pipette tips are vertical with the bottom of orifice plate as far as possible, and the distance of scratch can select the pipette tips of different model to adjust.Scratch is in same side
To in alignment.First: made " scratch " it is difficult to ensure that consistency, experimental error is big, disturbing factor is more, and manually
Scratch may scratch the coating on culture vessel surface, and then influence experimental result.Secondly, cell scar Healing Experiments need to connect
Effect of the continuous observation with assessment processing factor to cell migration, conventional method are in culture back preparatory marking observation position
It sets, but cell growth status can not be predicted before experiment, each observation point is also impossible to accomplish complete one under the microscope
Cause, and conventional method calculate be some position migration distance, can only obtain local cells migration data.
Summary of the invention:
The present invention is directed to the deficiency of existing cell scratch experiment technology, devises a kind of cell scar healing suitable for six orifice plates
Experimental provision can get circular cell scar, from the bulk migration situation of two dimension angular assessment cell.
Realize that technical scheme is as follows:
A kind of cell scar Healing Experiments device suitable for six orifice plates, including tablet (1), tablet top surface are provided with cross
Intersection construction (2), tablet (1) bottom surface are provided with cylindrical body (3), and cylindrical body (3) is correspondingly arranged in 4 of criss-cross construction
Below cross end (6).
Above-mentioned tablet is circle, and the crosspoint (5) of criss-cross construction (2) is located at the center location of round sheet object.
Above-mentioned criss-cross construction is length (L) 3.48cm, height (H) 1.0cm, thickness (T) 0.2cm；Round sheet object
Diameter (D) 1.88cm, thickness (T) 0.1cm.
Above-mentioned cylindrical body (3) height (H) 0.3cm, basal diameter (D) 0.2cm.
Above-mentioned roundel (1) is fixed on criss-cross construction (2), and cylindrical body (3) is fixed on roundel (1)；Cylinder
Body structure (3) is 4, is respectively corresponded below 4 cross end (6) midpoints of intersection construction (2).
Above-mentioned tablet (1) and criss-cross construction (2), cylindrical structure (3) are stainless steel material.
Beneficial effect
1. the present apparatus provides a kind of cell scar Healing Experiments suitable for six orifice plates, it is straight that 4 bottoms are obtained in a 6 orifice plates
Diameter is the round cell scar of 0.2cm, facilitates the cell scar healing state in observation whole visual field, Ke Yitong under the microscope
The gross area for calculating scar is crossed, cell scar healing rate is obtained.Avoid the current cell scratch experiment of cell that from can only observing scratch office
The healing state in portion region.(0.2cm is the observable maximum gauge under 40 × visual field of microscope).
2. the present apparatus is free of toxic effects to cell culture, can high temperature and pressure sterilize repeatedly, reuse, economical and practical ring
It protects.
3. the device that the present apparatus provides is suitable for 6 orifice plates, it is placed in 6 orifice plates before cell culture, cell is added, to cell
After adherent, after removing device, the round cell scar of neat in edge can be seen, can under the microscope dynamic observation cell healing
Overall condition can manufacture 4 round cell scars by once testing in six orifice plates, can get multiple experiment repeat numbers
According to helping to assess experimental result, greatly save experimental cost, save experimental period.
4. the scar Healing Experiments of various cells can be used for well using the present apparatus, and cell can be kept living well
Property.
Detailed description of the invention
Fig. 1 present apparatus side view (1- tablet, 2- criss-cross construction, 3- cylindrical structure)
Fig. 2 present apparatus is placed in top view in six orifice plates (1- roundel, 2- criss-cross construction, 3- cylindrical structure, 4- six
Orifice plate, 5- criss-cross construction crosspoint, 6- criss-cross construction cross end)
Fig. 3 present apparatus is placed in bottom view in six orifice plates (1- roundel, 2- criss-cross construction, 3- cylindrical structure, 4-
Six orifice plates, 6- criss-cross construction cross end)
Schematic diagram (3- round cell scar, six orifice plate of 4-) after Fig. 4 present apparatus is taken out
Fig. 5 pipette tips scratch is compared with the round scar of this cell scar device for healing
The healing state of 1,3,5 day round scar after Fig. 6 ATRA processing
Fig. 7 cell scar healing rate (*: p < 0.05)
Specific embodiment
The present invention is specifically described below by embodiment, it is necessary to which indicated herein to be, following embodiment is only used
It is further detailed in the present invention, should not be understood as limiting the scope of the invention, which is skilled in technique
Personnel can make some nonessential modifications and adaptations to the present invention according to foregoing invention content.
Embodiment 1
A kind of cell scar Healing Experiments device suitable for six orifice plates, including tablet (1), tablet top surface are provided with cross
Intersection construction (2), tablet (1) bottom surface are provided with cylindrical body (3), and cylindrical body (3) is correspondingly arranged in the ten of criss-cross construction
Below word end (6)；The tablet is circle, and the crosspoint (5) of criss-cross construction (2) is located at the center of circle position of round sheet object
It sets；The criss-cross construction is length (L) 3.48cm, height (H) 1.0cm, thickness (T) 0.2cm；The diameter of round sheet object
(D) 1.88cm, thickness (T) 0.1cm；Described cylindrical body (3) height (H) 0.3cm, basal diameter (D) 0.2cm；The disc-shaped
Object (1) is fixed on criss-cross construction (2), and cylindrical body (3) is fixed on roundel (1)；Cylindrical structure (3) is 4, respectively
Below corresponding 4 cross ends (6) for being located at intersection construction (2)；The tablet (1) and criss-cross construction (2), cylindrical knot
Structure (3) is stainless steel material.
This device is used for cell scar Healing Experiments detection all-trans retinoic acid (ATRA) to mouse tumor cell of liver
The effect of Hepal-6:
(1) it will be put into 6 orifice plates after present apparatus autoclave sterilization.
(2) by Hepal-6 cell with 2 × 106/ hole is seeded to 6 orifice plates, after 10 μm of ol/L concentration ATRA handle 3d, cell
Convergence degree takes out cell scar device close to 100%, the round cell scar that four diameters are 0.2cm is obtained, with manual pipette tips
Scratch compares, and the cell scar obtained under device is neat, and edge is uniform (Fig. 1).
(3) respectively at 0,1,3,5 day healing state in same position observation cell scar, measurement different parts scar is wide
Degree calculates healing rate, scar healing rate (%)=(0h scar area-X h scar area)/0h scratch area × 100%, experiment
It is repeated 3 times, as a result prompts ATRA that can effectively inhibit the migration of mouse tumor cell of liver Hepal-6, and the scar that this method calculates
Healing rate standard deviation is small, and experiment repetitive rate is high.
Claims (6)
1. a kind of cell scar Healing Experiments device suitable for six orifice plates, including tablet (1), tablet top surface are provided with ten
Word intersection construction (2), tablet (1) bottom surface are provided with cylindrical body (3), and cylindrical body (3) is correspondingly arranged in criss-cross construction
Below cross end (6).
2. being suitable for the cell scar Healing Experiments device of six orifice plates as described in claim 1, the tablet is round, ten
The crosspoint (5) of word intersection construction (2) is located at the center location of round sheet object.
3. being suitable for the cell scar Healing Experiments device of six orifice plates as claimed in claim 2, the criss-cross construction is
Length (L) 3.48cm, height (H) 1.0cm, thickness (T) 0.2cm；Diameter (D) 1.88cm of round sheet object, thickness (T)
0.1cm。
4. being suitable for the cell scar Healing Experiments device of six orifice plates as described in claim 1, cylindrical body (3) height
(H) 0.3cm, basal diameter (D) 0.2cm.
5. being suitable for the cell scar Healing Experiments device of six orifice plates as described in claim 1, the roundel (1) is fixed
In criss-cross construction (2), cylindrical body (3) is fixed on roundel (1)；Cylindrical body (3) is 4, respectively corresponds to be located at and intersect
Below 4 cross end (6) midpoints of structure (2).
6. the cell scar Healing Experiments device a method as claimed in any one of claims 1 to 5 for being suitable for six orifice plates, tablet (1) and ten
Word intersection construction (2), cylindrical structure (3) are stainless steel material.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811137312.1A CN109055218A (en) | 2018-09-17 | 2018-09-17 | A kind of cell circle scar Healing Experiments device for six orifice plates |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811137312.1A CN109055218A (en) | 2018-09-17 | 2018-09-17 | A kind of cell circle scar Healing Experiments device for six orifice plates |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN109055218A true CN109055218A (en) | 2018-12-21 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201811137312.1A Pending CN109055218A (en) | 2018-09-17 | 2018-09-17 | A kind of cell circle scar Healing Experiments device for six orifice plates |
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| CN (1) | CN109055218A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109055219A (en) * | 2018-09-17 | 2018-12-21 | 重庆医科大学附属儿童医院 | A kind of cell scar Healing Experiments device |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN203048919U (en) * | 2013-01-08 | 2013-07-10 | 复旦大学 | Stamp type experimental device for cell migration experiments |
| KR20180057157A (en) * | 2016-11-22 | 2018-05-30 | 경희대학교 산학협력단 | Method for forming wound and in vitro wound healing assay |
| CN209854173U (en) * | 2018-09-17 | 2019-12-27 | 重庆医科大学附属儿童医院 | Cell circular scar healing experimental device for six-hole plate |
| CN217173759U (en) * | 2022-03-19 | 2022-08-12 | 重庆医科大学附属儿童医院 | Culture container for transporting cells |
-
2018
- 2018-09-17 CN CN201811137312.1A patent/CN109055218A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN203048919U (en) * | 2013-01-08 | 2013-07-10 | 复旦大学 | Stamp type experimental device for cell migration experiments |
| KR20180057157A (en) * | 2016-11-22 | 2018-05-30 | 경희대학교 산학협력단 | Method for forming wound and in vitro wound healing assay |
| CN209854173U (en) * | 2018-09-17 | 2019-12-27 | 重庆医科大学附属儿童医院 | Cell circular scar healing experimental device for six-hole plate |
| CN217173759U (en) * | 2022-03-19 | 2022-08-12 | 重庆医科大学附属儿童医院 | Culture container for transporting cells |
Non-Patent Citations (1)
| Title |
|---|
| 顾其胜主编: "海藻酸盐基生物医用材料与临床医学", 上海：上海科学技术出版社, pages: 325 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109055219A (en) * | 2018-09-17 | 2018-12-21 | 重庆医科大学附属儿童医院 | A kind of cell scar Healing Experiments device |
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