CN105199941B - One kind removing cumulus cell device - Google Patents
One kind removing cumulus cell device Download PDFInfo
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- CN105199941B CN105199941B CN201510658335.7A CN201510658335A CN105199941B CN 105199941 B CN105199941 B CN 105199941B CN 201510658335 A CN201510658335 A CN 201510658335A CN 105199941 B CN105199941 B CN 105199941B
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- Prior art keywords
- small opening
- ware
- cumulus
- pin
- druming
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- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Expired - Fee Related
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- 210000001771 cumulus cell Anatomy 0.000 title claims abstract description 51
- 210000000130 stem cell Anatomy 0.000 claims abstract description 36
- 238000010009 beating Methods 0.000 claims abstract description 4
- 238000007664 blowing Methods 0.000 claims abstract description 4
- 150000001875 compounds Chemical class 0.000 claims description 25
- 210000000287 oocyte Anatomy 0.000 claims description 23
- 239000010410 layer Substances 0.000 claims description 17
- 238000004140 cleaning Methods 0.000 claims description 10
- 239000007788 liquid Substances 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 6
- 235000015097 nutrients Nutrition 0.000 claims description 6
- 239000012153 distilled water Substances 0.000 claims description 3
- 239000011521 glass Substances 0.000 claims description 3
- 239000011241 protective layer Substances 0.000 claims description 3
- 238000007790 scraping Methods 0.000 claims description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 3
- 238000000926 separation method Methods 0.000 claims description 2
- 230000006378 damage Effects 0.000 abstract description 3
- 208000027418 Wounds and injury Diseases 0.000 abstract description 2
- 208000014674 injury Diseases 0.000 abstract description 2
- 230000004083 survival effect Effects 0.000 abstract description 2
- 102000002322 Egg Proteins Human genes 0.000 description 7
- 108010000912 Egg Proteins Proteins 0.000 description 7
- 210000004681 ovum Anatomy 0.000 description 7
- 230000002611 ovarian Effects 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 230000018109 developmental process Effects 0.000 description 3
- 108010003272 Hyaluronate lyase Proteins 0.000 description 2
- 102000001974 Hyaluronidases Human genes 0.000 description 2
- 238000013461 design Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 230000000694 effects Effects 0.000 description 2
- 229960002773 hyaluronidase Drugs 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- 241000486406 Trachea Species 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000000903 blocking effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000035800 maturation Effects 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000008182 oocyte development Effects 0.000 description 1
- 230000002093 peripheral effect Effects 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 238000005201 scrubbing Methods 0.000 description 1
- 210000003437 trachea Anatomy 0.000 description 1
Classifications
-
- 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
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/06—Bioreactors or fermenters specially adapted for specific uses for in vitro fertilization
-
- 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
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N5/00—Undifferentiated human, animal or plant cells, e.g. cell lines; Tissues; Cultivation or maintenance thereof; Culture media therefor
- C12N5/0081—Purging biological preparations of unwanted cells
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- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Organic Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Chemical & Material Sciences (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Biomedical Technology (AREA)
- Microbiology (AREA)
- Biochemistry (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Sustainable Development (AREA)
- Molecular Biology (AREA)
- Clinical Laboratory Science (AREA)
- Cell Biology (AREA)
- Apparatus Associated With Microorganisms And Enzymes (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
Abstract
The invention discloses one kind removing cumulus cell device, including small opening ware, piping and druming pin, air bag；Small opening ware is circle, and ware bottom uniformly removes the small opening of cumulus cell, and a diameter of 90 μm~150 μm of small opening is provided with internal thread in hole；Piping and druming pin is provided with handle, and the needle point for blowing and beating pin is round and smooth, and piping and druming pin is connected by tracheae with air bag.Simple to operate, efficiency high of the invention, saves manpower；Mechanical removal cumulus cell, reduces the injury to egg mother cell；It is time-consuming short, improve egg mother cell survival rate.
Description
Technical field
The present invention relates to one kind removing cumulus cell device.
Background technology
Egg mother cell is commonly called as ovum, and egg mother cell is wrapped up during early development by cumulus cell, forms ovarian cumulus ovum female
Cell complexes, after after Oocyte Development maturation, the cumulus cell around egg mother cell completes its physiological function.In reproduction
In order to further investigate the genesis and development mechanism of egg mother cell, it is necessary to remove the ovarian cumulus around egg mother cell in many researchs such as development
Cell.At present, removing egg mother cell around cumulus cell be mainly by operate liquid in add hyaluronidase, hyalomitome
Sour enzyme is used as commercialization biological reagent, and its biologically active labile is higher to storage requirement requirement, easily right during digestion cumulus cell
Egg mother cell is damaged, also, is also required to mechanical artificial piping and druming after addition hyaluronidase, if piping and druming dynamics not enough may
Can cause easily take off cumulus cell, dynamics is excessive and egg mother cell can be caused to injure or dead；Artificial mechanism is taken to blow
Removing cumulus cell is beaten, at present due to lacking professional operation device, is operated than relatively time-consuming and less efficient, the operating time
Egg mother cell is more long in the extraneous operating environment time when long, easily causes egg mother cell mortality or influence ovum is female thin
Born of the same parents' later stage develops.For this problem, the present invention designs a new removing cumulus cell device to solve this problem.
The content of the invention
The technical problem to be solved in the present invention is to provide a kind of removing cumulus cell device.
The invention solves the problems that another technical problem be to provide a kind of removing cumulus cell method.
For removing cumulus cell device, the technical solution adopted by the present invention is that one kind removes cumulus cell device, including
Small opening ware, piping and druming pin, air bag；
Small opening ware is circle, and ware bottom uniformly removes the small opening of cumulus cell, a diameter of 90 μm of small opening~150 μm, small opening
Inside it is provided with internal thread；
Piping and druming pin is provided with handle, and the needle point for blowing and beating pin is round and smooth, and piping and druming pin is connected by tracheae with air bag.
Preferably, small opening diameter is respectively 150 μm, 120 μm, 90 μm, 3 kinds of small openings of different-diameter are from circular ware bottom
The center of circle is uniformly distributed from inside to outside from big to small along radius by diameter.
Preferably, the ware bottom of small opening ware is provided with the circular pit for connecting culture dish.
Preferably, a diameter of 66mm of small opening ware, a diameter of 60mm of circular pit at ware bottom, for connecting 60mm cultures
Ware.
Preferably, the tip diameter of piping and druming pin is set as 120 μm~160 μm.
Preferably, air bag jams on contracting air-generating for pin.
Preferably, tracheae includes outer layer and internal layer, outer layer is protective layer, and internal layer is snorkel, and the length of tracheae is 1.5
~2m.
Preferably, also including Cleaning tool made of plastic；Cleaning tool top is handle, and bottom is piston,
Connection in the middle of hand and piston；Piston diameter is adapted with the internal diameter of small opening ware, and piston ridge is provided with multi-layer groove.
Preferably, small opening ware is made up of glass.
For removing egg mother cell method, the technical solution adopted by the present invention is to comprise the following steps：
(1) nutrient solution is added in small opening ware, the cumulus oocyte compound that will be washed is moved into small opening ware, a front and rear left side
Small opening ware is rocked on the right side, and purpose is uniformly distributed cumulus oocyte compound；
(2) annular groove that will move into the small opening ware ware bottom of cumulus oocyte compound is placed in above a culture dish,
Appropriate operation liquid is added in culture dish；
(3) cumulus oocyte compound is put under microscope, the handle of hand-held piping and druming pin, pin is stepped on air bag, inhaled first
Enter one section of liquid, purpose cumulus oocyte compound is then found under the microscope, piping and druming pin and purpose ovarian cumulus ovum is female thin
Born of the same parents' compound, the small opening three alignment at small opening ware ware bottom, jam on air bag, and be blown into for purpose cumulus oocyte compound by piping and druming pin
The small opening of cumulus cell is removed, by the scraping retardation of concavo-convex screw thread staggeredly on small opening inwall so that be attached to ovum female
The cumulus cell of cell peripheral comes off and is separated from around egg mother cell, and the egg mother cell and part cumulus cell after separation are final
Fall into the culture dish being connected with small opening ware bottom, carry out subsequent operation；
(4) choose the immigration nutrient solution of the egg mother cell after having removed cumulus cell to be cultivated；
(5) drop in small opening ware is outwelled, adds distilled water cleaning standby.
The beneficial effects of the invention are as follows：
1st, simple to operate, efficiency high saves manpower；
2nd, mechanical removal cumulus cell, reduces the injury to egg mother cell；
3rd, short, raising egg mother cell survival rate is taken.
Brief description of the drawings
The present invention is further detailed explanation with reference to the accompanying drawings and detailed description.
Fig. 1 is the small opening ware front view of present invention removing cumulus cell device embodiment.
Fig. 2 is the small opening ware top view of present invention removing cumulus cell device embodiment.
Fig. 3 is the piping and druming needle construction schematic diagram of present invention removing cumulus cell device embodiment.
Fig. 4 is the Cleaning tool structural representation of present invention removing cumulus cell device embodiment.
Fig. 5 is the use schematic diagram of present invention removing cumulus cell device embodiment.
Marked in figure, 1- small opening wares, 2- macropores, 3- mesopores, 4- apertures, 5- syringe needles, 6- handles, 7- tracheaes, 8- air bags, 9-
Piston, 10- grooves, 11- handles, 12- culture dishes.
Specific embodiment
Fig. 5 is a kind of removing cumulus cell device, is made up of small opening ware, piping and druming pin, air bag.
Small opening ware 1 as shown in Figure 1 is the circular glass ware of diameter 66mm, and the ware bottom of small opening ware is provided with circular pit, circle
A diameter of 60mm of shape pit and be the same center of circle with ware bottom, circular pit is used to connect the culture dish of 60mm diameters.In small opening
Ware ware bottom uniformly removes the small opening of cumulus cell, and small opening is respectively the macropore 2 of 150 μm of the diameter, mesopore 3 of 120 μm of diameter, straight
The aperture 4 that 90 μm of footpath, it is equal from inside to outside from big to small that 3 kinds of small openings of different-diameter press diameter from the center of circle at circular ware bottom along radius
Even distribution (Fig. 2).
A diameter of 150 μm of macropore is suitable for egg mother cell diameter (including oolemma around egg mother cell) and exists in small opening
Between 120 μm~150 μm, the diameter that a diameter of 120 μm of mesopore is suitable for egg mother cell is (including transparent around egg mother cell
Band) between 90 μm~120 μm, a diameter of 90 μm diameters (including oolemma around egg mother cell) for being suitable for egg mother cell exist
Between 60 μm~90 μm.At present, common experimental animal egg mother cell diameter (including oolemma around egg mother cell) 60 μm~
Between 150 μm.
Internal thread is provided with small opening, the screw thread in small opening has the work(of mechanical removal egg mother cell surrounding cumulus cells
Can, concavo-convex Design of Screw Thread staggeredly can efficiently remove cumulus cell around egg mother cell by blocking screening mode.
As shown in figure 3, piping and druming pin is provided with handle 6, handle is anterior equipped with piping and druming pin syringe needle 5, and the needle point of syringe needle is round and smooth, prevents
Hurt egg mother cell.Handle 6 is connected by tracheae 7 with air bag 8.
The diameter of syringe needle 5 for blowing and beating pin is set as 120 μm~160 μm, it is therefore an objective in piping and druming cumulus oocyte compound mistake
Cheng Zhong, can completely cover cumulus oocyte compound and blow and beat again, prevent cumulus oocyte compound in the middle of piping and druming process
It is blown to elsewhere.
Air bag 8 jams on contracting air-generating for pin.
Tracheae 7 includes outer layer and internal layer, and outer layer is protective layer, and internal layer is snorkel, and the length of tracheae is 1.5~2m.
In piping and druming pin using preceding, first with piping and druming pin one section of liquid of inspiration, it is therefore an objective to keep cumulus oocyte compound to exist
It is maintained in liquid during piping and druming, reduces and damage.
Cleaning tool as shown in Figure 4 is made of plastic.Cleaning tool top be handle 11, bottom be piston 9, handle with
Connection in the middle of piston.
Piston diameter is 66~68mm, is adapted with the internal diameter with small opening ware, just somewhat firmly just can fill in small opening ware 1
Inner chamber, to be scrubbed.Multi-layer groove 10 is provided with piston periphery, the effect of groove is to reduce piston ridge and leakage
The contact surface of hole ware, to reduce friction, and there is multi-layer groove small contact layer can play the more preferable effect of scrubbing.
The method that cumulus cell is removed using said apparatus, is comprised the following steps：
(1) nutrient solution is added in small opening ware, the cumulus oocyte compound that will be washed is moved into small opening ware, a front and rear left side
Small opening ware is rocked on the right side, and purpose is uniformly distributed cumulus oocyte compound；
(2) the ware bottom that will move into the small opening ware of cumulus oocyte compound is placed in a culture dish of a diameter of 60mm 12
Above, allow the circular pit at the ware bottom of small opening ware 1 to be just stuck in the upper edge of culture dish 12, then added in 60mm culture dishes 12
Appropriate operation liquid；
(3) cumulus oocyte compound is put under microscope, the handle of hand-held piping and druming pin, pin is stepped on air bag, inhaled first
Enter one section of liquid, purpose cumulus oocyte compound is then found under the microscope, piping and druming pin and purpose ovarian cumulus ovum is female thin
Born of the same parents' compound, the small opening three alignment at small opening ware ware bottom, jam on air bag, and be blown into for purpose cumulus oocyte compound by piping and druming pin
Removing egg mother cell small opening, by the scraping retardation of concavo-convex screw thread staggeredly on small opening inwall so that cumulus cell and ovum
Mother cell is separated, and is eventually fallen into the 60mm culture dishes being connected with small opening ware bottom, carries out subsequent operation；
(4) choose the egg mother cell after having removed cumulus cell and move into nutrient solution culture；
(5) drop in small opening ware is outwelled, adds distilled water and cased after being cleaned with Cleaning tool preservation, used time alcohol
Cleaning, dries and can be used.
Invention described above implementation method, is not intended to limit the scope of the present invention..It is any in the present invention
Spirit and principle within modification, equivalent and the improvement made etc., should be included in claim protection model of the invention
Within enclosing.
Claims (10)
- It is 1. a kind of to remove cumulus cell device, it is characterised in that：Including small opening ware, piping and druming pin, air bag；The small opening ware is circle, and ware bottom uniformly removes the small opening of cumulus cell, a diameter of 90 μm~150 μm of the small opening, Internal thread is provided with small opening；The piping and druming pin is provided with handle, and the needle point for blowing and beating pin is round and smooth, and piping and druming pin is connected by tracheae with air bag.
- It is 2. according to claim 1 to remove cumulus cell device, it is characterised in that：The small opening diameter be respectively 150 μm, 120 μm, 90 μm, it is uniform from inside to outside from big to small that 3 kinds of small openings of different-diameter press diameter from the center of circle at circular ware bottom along radius Distribution.
- It is 3. according to claim 1 to remove cumulus cell device, it is characterised in that：The ware bottom of the small opening ware be provided with for Connect the circular pit of culture dish.
- It is 4. according to claim 3 to remove cumulus cell device, it is characterised in that：A diameter of 66mm of the small opening ware, The a diameter of 60mm of circular pit at ware bottom, to facilitate connection 60mm culture dishes.
- It is 5. according to claim 1 to remove cumulus cell device, it is characterised in that：The tip diameter setting of the piping and druming pin It is 120 μm~160 μm.
- It is 6. according to claim 1 to remove cumulus cell device, it is characterised in that：The air bag jams on contracting aerogenesis for pin Formula.
- It is 7. according to claim 1 to remove cumulus cell device, it is characterised in that：The tracheae includes outer layer and internal layer, Outer layer is protective layer, and internal layer is snorkel, and the length of tracheae is 1.5~2m.
- It is 8. according to claim 1 to remove cumulus cell device, it is characterised in that：Also work is scrubbed including made of plastic Tool；The Cleaning tool top is handle, and bottom is piston, connection in the middle of handle and piston；Piston diameter is interior with small opening ware Footpath is adapted, and piston ridge is provided with multi-layer groove.
- It is 9. according to claim 1 to remove cumulus cell device, it is characterised in that：The small opening ware is made up of glass.
- 10. using the removing cumulus cell method of the removing cumulus cell device described in claim 1, it is characterised in that including with Lower step：(1) nutrient solution is added in small opening ware, by washed cumulus oocyte compound immigration small opening ware, is all around shaken Dynamic small opening ware, purpose is uniformly distributed cumulus oocyte compound；(2) annular groove that will move into the small opening ware ware bottom of cumulus oocyte compound is placed in above a culture dish, in culture Appropriate operation liquid is added in ware；(3) cumulus oocyte compound is put under microscope, the handle of hand-held piping and druming pin, pin steps on air bag, one is sucked first Section liquid, then finds purpose cumulus oocyte compound under the microscope, and piping and druming pin is answered with purpose cumulus oocyte Compound, the small opening three alignment at small opening ware ware bottom, jam on air bag, and purpose cumulus oocyte compound is blown into removing by piping and druming pin The small opening of cumulus cell, by the scraping retardation of concavo-convex screw thread staggeredly on small opening inwall so that be attached to egg mother cell The cumulus cell of surrounding comes off and is separated from around egg mother cell, and egg mother cell and part cumulus cell after separation eventually fall into In the culture dish being connected with small opening ware bottom, subsequent operation is carried out；(4) choose the immigration nutrient solution of the egg mother cell after having removed cumulus cell to be cultivated；(5) drop in small opening ware is outwelled, adds distilled water cleaning standby.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510658335.7A CN105199941B (en) | 2015-10-10 | 2015-10-10 | One kind removing cumulus cell device |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510658335.7A CN105199941B (en) | 2015-10-10 | 2015-10-10 | One kind removing cumulus cell device |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105199941A CN105199941A (en) | 2015-12-30 |
| CN105199941B true CN105199941B (en) | 2017-05-31 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201510658335.7A Expired - Fee Related CN105199941B (en) | 2015-10-10 | 2015-10-10 | One kind removing cumulus cell device |
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| Country | Link |
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| CN (1) | CN105199941B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110184174A (en) * | 2019-07-12 | 2019-08-30 | 贵州省人民医院 | A kind of multipurpose cell and particle screen needle construction |
Family Cites Families (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN201150570Y (en) * | 2007-11-30 | 2008-11-19 | 许玉刚 | Mouth saccule type ovum peeling device |
| CN201309925Y (en) * | 2008-04-26 | 2009-09-16 | 刘雨生 | Granular cell dismantling tray used in external fertilization embryo transplantation technology |
| WO2010141320A1 (en) * | 2009-06-01 | 2010-12-09 | Mariposa Biotechnology, Inc. | Device for removing cumulus from oocytes |
| CN104055560B (en) * | 2014-07-06 | 2016-03-02 | 长沙傲图生物科技有限公司 | A kind of electronic stripping ovum device and stripping ovum method thereof |
| CN204246224U (en) * | 2014-11-24 | 2015-04-08 | 遵义医学院 | A kind of novel stripping egg apparatus |
| CN204434608U (en) * | 2015-02-03 | 2015-07-01 | 中国农业科学院特产研究所 | A kind of ovocyte takes off ovarian cumulus pin |
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