CN106190840B - Embryo Culture device and its bracket - Google Patents
Embryo Culture device and its bracket Download PDFInfo
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- CN106190840B CN106190840B CN201610707688.6A CN201610707688A CN106190840B CN 106190840 B CN106190840 B CN 106190840B CN 201610707688 A CN201610707688 A CN 201610707688A CN 106190840 B CN106190840 B CN 106190840B
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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
- C12M21/00—Bioreactors or fermenters specially adapted for specific uses
- C12M21/06—Bioreactors or fermenters specially adapted for specific uses for in vitro fertilization
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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
- C12M23/38—Caps; Covers; Plugs; Pouring means
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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
- C12M23/48—Holding appliances; Racks; Supports
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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
- C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
- C12M41/12—Means for regulation, monitoring, measurement or control, e.g. flow regulation of temperature
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- 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
- C12N5/06—Animal cells or tissues; Human cells or tissues
- C12N5/0602—Vertebrate cells
- C12N5/0603—Embryonic cells ; Embryoid bodies
- C12N5/0604—Whole embryos; Culture medium therefor
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Abstract
The present invention relates to a kind of Embryo Culture devices, including a culture tank, culture tank is equipped with lid, culture tank and lid thread fitting, and the lid of culture tank, which closes, sets air hole at screw thread, bracket placement platform is equipped in the culture tank, the position of bracket placement platform is higher than the position of air hole, and bracket is placed on bracket placement platform, and the bracket is in hollow out cellular, the bracket includes one block of porous plate, and bulge loop is set on porous plate.After apparatus of the present invention, quantity required of the Test-Tube Baby Lab to carbon dioxide incubator is significantly reduced, significantly saves the space in laboratory and the usage amount of carbon dioxide and nitrogen;Improve safety, stability and the embryo quality of human embryos in vitro culture and the safety in laboratory.
Description
Technical field
The present invention relates to a kind of Embryo Culture device and its brackets, belong to IVF-ET cycle technical field.
Background technology
The mankind IVF-ET cycle technology (InVitro-FertilizationandEmbryoTransfer,
IVF-ET) developed more than 30 years, had been achieved for larger progress.Current inseminatio externalis and Embryo Culture are in titanium dioxide
It is completed in carbon incubator, relies on the chemical composition in the carbon dioxide and culture medium of stationary temperature and constant density in incubator
It interacts and exchanges, to maintain smart ovum insemination and the physiological environment needed for embryo growth.But in existing culture systems no matter
It is on hardware device (culture apparatus or incubator) or all there are some defects or deficiencies in terms of culture medium.With regard to hardware
Speech, existing culture apparatus (incubator) are an open systems, the gas in carbon dioxide incubator need constantly with
Outside case i.e. embryo experiments indoor air exchange could maintain incubator interior or device in gas balance and stability, therefore in this way
Open system there are following clearly disadvantageous or defects:First, the gas concentration and temperature in device are actually in one
The dynamic equilibrium state of kind, gas concentration and temperature fluctuate in certain range, and the size of fluctuation then depends on device or training
Support the enabling frequency of the model of case, quality and incubator;Second is that test indoor volatile organic matter, harmful physical and chemical factor such as
Ozone etc. can be entered by this open gas communication system in carbon dioxide incubator, lead to the damage of embryo,
It is even dead;The third is when carbon dioxide incubator opens the door, the gas in case largely overflows and inevitably results in culture
It fluctuates to gas concentration and high temperature in case, with the increase of door opening times, this big ups and downs cause what embryo injured
Degree also increases therewith.
To solve this technical problem, I discloses one in the patent of invention CN201410284830.1 applied before
Kind is inseminated for mankind's ovum with sperm in vitro and the culture apparatus of early embryo development, achieves extraordinary Embryo Culture effect
Fruit has simultaneously obtained practical application.But this equipment is more expensive, the poor hospital of condition is more difficult to undertake equipment purchase expense,
And at present age for marriage gradually increase, the above the average age for marriage Mr. and Mrs success rate that is pregnant naturally is relatively low, infertile to solve the problems, such as, satisfaction
Each woman does the dream of mother, and it is necessary to promote for IVF-ET cycle technology.
Invention content
Above-mentioned condition based on the prior art, the purpose of the present invention is to provide a kind of Embryo Culture devices, which can
It is closed after inflation, it is then then transferred in common Embryo Culture case and carries out Embryo Culture, ensure Embryo Culture quality, and should
Apparatus structure is simple, easily make, low manufacture cost is not only easy to use, and the culture effect for embryo, safety and
Stability has highly significant raising.It can wide popularization and application.
To achieve these goals, present invention employs following technical proposals.
A kind of Embryo Culture device, which is characterized in that including a culture tank, culture tank is equipped with lid, culture tank and lid
Sub- thread fitting, the lid of culture tank close and air hole are set at screw thread, and bracket placement platform is equipped in the culture tank, and bracket is placed
Bracket is placed on platform, the bracket is in hollow out cellular.
Further preferably, the culture tank is assembled by outer tank and inner canister, there are heat-insulation chamber between outer tank and inner canister, every
Hot chamber keeps low pressure, i.e., with certain vacuum degree or vacuumizes, such culture tank can play the work of heat-insulation and heat-preservation
With;Similarly, it is also provided with heat-insulation chamber in the lid.The culture tank and the shape of lid are regular polygon prism, are screwed in this way
It is convenient.
Further preferably, outer tank and inner canister are threadedly coupled and set sealing ring.
Further preferably, the bracket includes one block of porous plate, and bulge loop is set on porous plate.
Further preferably, it is distributed hole on the side wall of the bulge loop.
Further preferably, the porous plate is one piece of Multi-angular square, and the center of Multi-angular square opens up central through hole, Multi-angular square it is each
A angle connects an annulus jointly, is distributed hole on Multi-angular square.
Further preferably, the culture tank and the material of bracket are stainless steel or other metals or the material nontoxic to embryo
Material.
During Embryo Culture, bracket is placed in culture tank, (rear referred to as to cultivate equipped with reproduction cell, fertilized eggs or embryo
Object) culture dish be placed on bracket, cover the lid of culture tank, Gai Heshi, lid not cover the air hole of culture tank, so
After be positioned in carbon dioxide incubator and balance 60~120 minutes, screw lid, closed air hole is transferred to the device general
It is cultivated in logical Embryo Culture case, until developing until the required stage.
The present invention also provides a kind of Embryo Culture brackets, and including one block of porous plate, bulge loop is set on porous plate, described convex
Hole is distributed on the side wall of ring.
Further preferably, one annulus of periphery connection of porous plate.
Further preferably, the porous plate is one piece of Multi-angular square, such as Square consisting of two isosceles right-angled triangles, five gussets, the center of Multi-angular square open up
Central through hole, each angle of Multi-angular square connect an annulus, are distributed hole on each gusset jointly.Why using Multi-angular square and
Center opens up central through hole, is because such bracket not only makes the gas circulation of two sheaf spaces above and below bracket splendid, moreover it is possible to
Facilitate placement and taking-up, and bracket from falling or overturning can also be prevented, improve the safety of culture.
The bracket of the present invention, after the culture dish equipped with culture is placed on the bulge loop of bracket, the gas in culture tank can
Across the hole free flow of porous plate so that Embryo Culture environment is more uniformly distributed stabilization, ensures that embryonic development is good.
11. the present invention also provides a kind of new Embryo Culture methods, it is characterised in that:Culture is the two of suitable concentration
In the gaseous environment of carbonoxide or/and nitrogen gas concn after short time balance, independent space is confined to, then need not reuse two
Carbonoxide or/and nitrogen, it is only necessary under conditions of steady temperature (such as 36.8-37 degrees Celsius) is provided, continue pedestrian's class fertilized eggs or
The in vitro culture of embryo;
Wherein:The device carbon dioxide incubator that is of gaseous environment equilibrium condition is provided or can be filled with the two of suitable concentration
Carbonoxide or the container with nitrogen;
Wherein:The device that closed separate space is provided be Embryo Culture device described in claim 1-4 any one or
Other sealable containers.
This method has jumped out the mindset that embryo can only cultivate in carbon dioxide incubator in Traditional Thinking, and rises
Good culture effect is arrived.
Advantages of the present invention:First, culture or standing time of the culture in carbon dioxide incubator is greatly reduced, from
And the quantity of carbon dioxide incubator and the usage amount of carbon dioxide and nitrogen can be greatly decreased, not only save laboratory sky
Between, also improve the safety of embryo experiments room.2nd, compared to carbon dioxide incubator cultivation, the present invention is because using this
It is positioned in a manner of closed culture in common insulating box after the kind good carbon dioxide of mode short time inner equilibrium and continues to cultivate, embryo
Gas and temperature in culture apparatus are capable of the constancy of height, will not change, different cultures as incubator opens the door
Between will not interfere with each other, except except culturing room for a long time power off in addition to external environment any variation or accident (such as laboratory disappears
Ultraviolet light of poison etc.) cultivation is not interfered with as a result, Embryo Culture quality is not interfered with, so as to be greatly improved embryo's
The safety and stability of training quality and in vitro culture.
The common Embryo Culture case of Embryo Culture device cost of manufacture low combination of the present invention can play splendid embryo's training
Effect is supported, suitable for promoting.
Description of the drawings
Fig. 1 is the schematic diagram of culture tank.
Fig. 2 is the schematic diagram of bracket.
Specific embodiment
In order to make it easy to understand, the present invention is further clarified below in conjunction with the accompanying drawings.
With reference to Fig. 1, a kind of Embryo Culture device, including a culture tank 1, culture tank 1 is equipped with lid 3, culture tank 1 and lid
Sub 3 thread fittings, the lid of culture tank 1 close and air hole 12 are set at screw thread, and bracket placement platform 11 is equipped in the culture tank 1,
Bracket 2 is placed on bracket placement platform 11, the bracket 2 is in hollow out cellular.The culture tank 1 and the shape of lid 2 is just
Polygon-prism conveniently screws.
With reference to Fig. 2, the bracket includes one piece of five gusset 21, and the center of five gussets 21 opens up central through hole, five gussets 21
Five angles, one annulus 23 of connection jointly, be distributed hole on five gussets 21.Bulge loop 22, the bulge loop 22 are set on five gussets 21
Side wall on be distributed hole.The diameter of the bulge loop 22 is bigger than the diameter of central through hole, forms a ring in bulge loop 22 in this way
Shape supporting plate, if using big culture dish, big culture dish is placed on bulge loop 22, and the height of big culture dish is higher than 1 tank mouth of culture tank,
It is convenient that culture dish is picked and placeed in this way;If using small culture dish, small culture dish is placed on the annular supporting plate in bulge loop 22, bulge loop 22
Hole and central through hole on side wall can play permeation functions.Because there is bulge loop 22, and also opened up on the side wall of bulge loop 22
Hole can place the culture dish of two kinds of specifications.
The bracket 2 sets annulus 23, and the shape of annulus 23 coordinates with the internal diameter size of culture tank 1, so that bracket 2
It will not be overturn due to shaking.The bracket 2 is porous plate, and sets bulge loop 22, after culture dish is placed on bulge loop 22, culture
Gas in tank 1 can ensure that culture environment is uniform and stable, in bulge loop by central through hole and other small hole free flows
Hole is also opened up on 22 side wall, further improves gas flow effect.
Further, the culture tank 1 is assembled by outer tank and inner canister, there are heat-insulation chamber between outer tank and inner canister, and
Low pressure is kept, i.e., with certain vacuum degree or is vacuumized, such culture tank 1 can play the role of heat-insulation and heat-preservation,
During Embryo Culture, it is more advantageous to preserving gas and temperature in culture tank 1 and stablizes.During actual fabrication, outer tank and inner canister are threadedly coupled
And sealing ring is set, other modes can also be used to make the culture tank 1 with vacuum chamber, the material of culture tank 1 is using stainless
Steel is other to embryo's innocuous materials.Heat-insulation chamber is also provided in the lid 3, plays the role of heat-insulation and heat-preservation.
During Embryo Culture, bracket 2 is placed in culture tank 1, and the culture dish equipped with culture is placed on carrier 2 or held in the palm
In frame, the lid 3, Gai Heshi of culture tank 1 is covered, Embryo Culture device is placed in carbon dioxide incubator, lid 3 not cover
The firmly air hole 12 of culture tank 1, balance screw lid 3 after 60~120 minutes, then closed air hole 12 shifts the device
It is cultivated into common insulating box, keeps constant temperature.It has the following advantages:First, multiple hairs can be placed in common insulating box
Bright Embryo Culture device, that has saved carbon dioxide incubator uses space and quantity, and improving carbon dioxide incubator makes
With efficiency, reduce carbon dioxide usage amount.2nd, compared to carbon dioxide incubator cultivation, the present invention is because using the short time
The mode of closed culture after the good carbon dioxide of inner equilibrium, gaseous environment and temperature in Embryo Culture device can keep height
Constancy will not change as incubator opens the door, will not be interfered with each other between different cultures, except culturing room is disconnected for a long time
Any variation of external environment other than electricity or accident (such as ultraviolet light of laboratory disinfection) do not interfere with cultivation as a result, not
Embryo Culture quality can be influenced, so as to which the safety of the training quality of embryo and in vitro culture and stabilization be greatly improved
Property.The Embryo Culture device of the present invention, by it was verified that with extremely excellent Embryo Culture effect, can with wide popularization and application,
Convenient for improving the technical merit and service quality of inseminatio externalis art.With
The application method of Embryo Culture device of the present invention and Embryo Culture method:
1st, device is packed into 50-100ml high purity waters using preceding in it, and the super of Embryo Culture rank is preferred in the person of having ready conditions
Pure water, with the saturated humidity in holding meanss;
2nd, using it is preceding be first positioned in carbon dioxide incubator it is spare overnight.It is special that a carbon dioxide incubator can be flowed out
The present apparatus is placed, so as to spare at any time.
3rd, the present apparatus in stand-by state is taken out, twists the lid off, the culture dish equipped with culture is put in a device
On bracket, be twisted lid, but should not closing device side air hole, be then placed in another carbon dioxide incubator and balance not
Less than 60 minutes.Lid is further tightened after balance, to ensure the complete complete closure of side stomata, is fully sealed so that device is in
State, be then transferred directly to continue to cultivate in 36.9-37 DEG C of common insulating box.
4th, each common insulating box can at least prevent the 10-20 present apparatus, be equivalent to the incubator 10-20 of equal volume
The usage amount of platform, and effect is more preferable, but its carbon dioxide for using and nitrogen amount only have the percent of carbon dioxide incubator
One or so.
Using effect data:
During in June, -2016 in July, 2014:Normal human subject fertilized eggs are co-cultured in conventional carbon dioxide incubator
450 pieces of son, culture form excellent spilting of an egg embryo (6-12 cells 1 or 2 grades) 238, more than 6 cells 3 grades of embryos 80 after 66-72 hours
Piece, 5 non-spilting of an egg, excellent spilting of an egg embryogenesis rate 53.5% shares 245 available embryos's (1-3 grades of 6-12 cells) and continues blastaea
Culture, obtains 174 pieces of blastaea, wherein forming 33 pieces of top blastaea (3-6AA grades), Blastocyst formation rate 71.0%, top Blastocyst formation
Rate is 19.0%;Formed excellent after 66-72 hours using 5320 pieces of present apparatus culture normal human subject fertilised egg, culture in the same period
Spilting of an egg embryo (classification is same as above) 3424, more than 6 cells 3 grades of 685 pieces of embryos, 53 non-spilting of an egg, excellent spilting of an egg embryogenesis rate
64.4%, it shares 3987 available embryos's (1-3 grades of 6-12 cells) and continues blastocyst culture, obtain 2910 pieces of blastaea, wherein forming top
1252 pieces of grade blastaea (3-6AA grades), Blastocyst formation rate 73.0%, top Blastocyst formation rate are 43.0%.Preliminary experiment results table
It is bright, compared with traditional carbon dioxide incubator, use its excellent spilting of an egg embryogenesis rate and top Blastocyst formation rate after the present apparatus
There is extremely significant raising.
Claims (3)
1. a kind of Embryo Culture device, which is characterized in that including a culture tank, culture tank is equipped with lid, culture tank and lid
Coordinate, air hole at the lid conjunction of culture tank is set, bracket placement platform is equipped in the culture tank, is placed on bracket placement platform
Bracket, the bracket are in hollow out cellular, and the bracket includes one block of porous plate, bulge loop, the culture tank are set on porous plate
There are heat-insulation chambers for inside;There are cavitys for cover inside, and after culture tank tightens closure with lid, inner space becomes confined space;
The porous plate is one piece of Multi-angular square, and the center of Multi-angular square opens up central through hole, and each angle of Multi-angular square connects a circle jointly
Ring is distributed hole on Multi-angular square;The diameter of the bulge loop is bigger than the diameter of central through hole, and an annular is formed in this way in bulge loop
Supporting plate.
2. Embryo Culture device according to claim 1, it is characterised in that:Hole is distributed on the side wall of the bulge loop.
A kind of 3. Embryo Culture method, it is characterised in that:Culture is in the carbon dioxide of suitable concentration or/and the gas of nitrogen gas concn
In body environment after short time balance, independent space is confined to, then need not reuse carbon dioxide or/and nitrogen, it is only necessary to carry
Under conditions of steady temperature, continue pedestrian's class fertilized eggs or the in vitro culture of embryo;
Wherein:The device for providing gaseous environment equilibrium condition is carbon dioxide incubator or the carbon dioxide that can be filled with suitable concentration
Or container or equipment with nitrogen;
Wherein:The device for providing closed separate space is the Embryo Culture device described in claims 1 or 2.
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CN106190840B true CN106190840B (en) | 2018-07-10 |
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Families Citing this family (5)
Publication number | Priority date | Publication date | Assignee | Title |
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CN107058099B (en) * | 2017-04-24 | 2023-06-27 | 中国农业科学院特产研究所 | Embryo culture and transportation device |
CN107022484A (en) * | 2017-05-27 | 2017-08-08 | 余裕炉 | A kind of Embryo Culture unit and preparation method thereof |
CN109251860A (en) * | 2018-10-17 | 2019-01-22 | 北京大学深圳医院 | Reproductive center laboratory embryo breeding utensil |
CN109401971B (en) * | 2018-12-28 | 2024-01-30 | 江苏省人民医院(南京医科大学第一附属医院) | Multifunctional automatic independent/combined co-culture device |
CN110042057A (en) * | 2019-05-07 | 2019-07-23 | 遵义医学院附属医院 | It is a kind of for external seminal fluid collecting and the device of Embryo Culture |
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