CN109486677A - A kind of Embryo Culture position limiting fence - Google Patents

A kind of Embryo Culture position limiting fence Download PDF

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Publication number
CN109486677A
CN109486677A CN201811513114.0A CN201811513114A CN109486677A CN 109486677 A CN109486677 A CN 109486677A CN 201811513114 A CN201811513114 A CN 201811513114A CN 109486677 A CN109486677 A CN 109486677A
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CN
China
Prior art keywords
position limiting
embryo
limiting fence
culture
embryo culture
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CN201811513114.0A
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Chinese (zh)
Inventor
郭镇华
吕蕾
杨云春
付博
刘自广
汪亮
彭福刚
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Institute of Animal Husbandry of Heilongjiang Academy of Agricultural Sciences
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Institute of Animal Husbandry of Heilongjiang Academy of Agricultural Sciences
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Priority to CN201811513114.0A priority Critical patent/CN109486677A/en
Publication of CN109486677A publication Critical patent/CN109486677A/en
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    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS 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
    • C12M21/00Bioreactors or fermenters specially adapted for specific uses
    • C12M21/06Bioreactors or fermenters specially adapted for specific uses for in vitro fertilization
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS 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
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/02Form or structure of the vessel
    • C12M23/12Well or multiwell plates
    • CCHEMISTRY; METALLURGY
    • C12BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12MAPPARATUS 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
    • C12M23/00Constructional details, e.g. recesses, hinges
    • C12M23/34Internal compartments or partitions

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  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Chemical & Material Sciences (AREA)
  • Zoology (AREA)
  • Genetics & Genomics (AREA)
  • Sustainable Development (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Biomedical Technology (AREA)
  • Clinical Laboratory Science (AREA)
  • Molecular Biology (AREA)
  • Apparatus Associated With Microorganisms And Enzymes (AREA)

Abstract

The invention discloses a kind of Embryo Culture position limiting fences, the production method of the Embryo Culture position limiting fence is specifically includes the following steps: S1, the material of Embryo Culture position limiting fence selects, it chooses polystyrene or other medical materials carries out the production of Embryo Culture position limiting fence, S2, the size design of Embryo Culture position limiting fence, according to embryo's diameter within the scope of 180-220um, each separation long 300um of lattice inside dimension can be designed, wide 300um, high 200um, each separation lattice thickness 50um, four frame thickness 0.6mm of edge, latticed Embryo Culture position limiting fence can be obtained, S3, the logo design of Embryo Culture position limiting fence, the present invention relates to Embryo engineering technology fields.The Embryo Culture position limiting fence, the good Embryo Culture position limiting fence of production can be achieved effectively to overcome single embryo in vitro in incubation the phenomenon that depauperation, simplify operating procedure simultaneously, achieve the purpose that ensure to distinguish embryonic origin well, to greatly facilitate the Embryo Culture work of people.

Description

A kind of Embryo Culture position limiting fence
Technical field
The present invention relates to Embryo engineering technology field, specially a kind of Embryo Culture position limiting fence.
Background technique
Embryo engineering refers to a variety of micromanipulations carried out to animal body early embryo or gamete and processing technique, including body Technologies, many technologies of embryo engineering such as outer fertilization, embryo transfer, embryo separating transplanting, embryonic stem cell culture really exist Under conditions in vitro, to the simulated operation that animal natural fertilization and early embryonic development condition carry out, embryo engineering can further be dug The breeding potential of animal is dug, is the mass propagation of superior animals, the race of rare animal, which continues, provides effective solution, The fields such as animal husbandry and pharmacy industry play an important role, and there is bright application prospect to commonly use embryo in embryology research Tire in vitro culture, but it is commonly encountered single embryo during the test in vitro in incubation the phenomenon that depauperation, cause to send out Termination is educated, has research to think, mainly since single embryo can not exchange signal factor, polyembryony tire co-incubation with other embryos Although can overcome the problems, such as this, embryonic origin can not be distinguished after culture.
It has been reported that at present and is first laid with one layer of agar inside culture plate, several duck eyes are then dug out on agar, are divided Every culture, because agar, which must be heated to 100 degrees Celsius, to be melted, culture solution is otherwise therein no more than 50 degrees Celsius Irreversible variation occurs at branch, the disadvantage is that it is complicated for operation, and the PBS solution for configuring agar influences whether culture solution Ingredient, and the present invention can effectively overcome single embryo in vitro in incubation the phenomenon that depauperation, simplify operation step Suddenly, it is ensured that distinguish embryonic origin.
Summary of the invention
(1) the technical issues of solving
In view of the deficiencies of the prior art, the present invention provides a kind of Embryo Culture position limiting fences, solve existing embryo's training It supports complicated for operation, cannot effectively overcome single embryo in vitro in incubation the phenomenon that depauperation, being unable to reach ensures point The problem of clear embryonic origin purpose.
(2) technical solution
In order to achieve the above object, the present invention is achieved by the following technical programs: a kind of Embryo Culture position limiting fence, it is described The production method of Embryo Culture position limiting fence specifically includes the following steps:
The material selection of S1, Embryo Culture position limiting fence, choose polystyrene or other medical materials carry out Embryo Culture limit The production of bit field;
The size design of S2, Embryo Culture position limiting fence can design each according to embryo's diameter within the scope of 180-220um A separation lattice inside dimension long 300um, wide 300um, high 200um, each separation lattice thickness 50um, four frame thickness of edge Latticed Embryo Culture position limiting fence can be obtained in 0.6mm;
The logo design of S3, Embryo Culture position limiting fence mark English alphabet, indulge and sit at each culture lattice abscissa Arabic numerals are marked at mark, so each culture lattice has unique double figures to identify;
The liquid fluidity design of S4, Embryo Culture position limiting fence, four walls of each culture lattice, using laser or plasma Technology is punched, and each bore dia is 50um, and the quantity and size in hole can be adjusted according to the size of embryo;
S5, manufactured Embryo Culture position limiting fence will be surface-treated, and Embryo Culture position limiting fence is passed through process of surface treatment Making the surface of Embryo Culture position limiting fence has hydrophilic or hydrophobicity;
S6, manufactured Embryo Culture position limiting fence will just can be used by ethylene oxide sterilizing or radiosterilization, make embryo Tire culture position limiting fence surface is in germ-free condition;
Embryo Culture position limiting fence is put into the hole of four well culture plates by the use of S7, Embryo Culture position limiting fence, adjusts position Set the center for occuping hole;
S8, then 500ul culture solution is injected into the hole of four well culture plates, pays attention to not making Embryo Culture position limiting fence floating It rises, if having air in the four walls cavity of culture lattice, micropipettor pressure-vaccum repeatedly can be used, be removed;
S9, will be above the culture solution in the careful hole for being injected into four well culture plates of 300ul mineral oil;
S10, by the cumulus oocyte compound of acquirement either fertilized embryo, cleaned three times with culture solution, be put into embryo It is cultivated in the culture lattice of tire culture position limiting fence.
Preferably, the size of Embryo Culture position limiting fence can be adjusted according to the stage of development of embryo in the step S2, expanded The long 600um of Embryo Culture position limiting fence, wide 600um, high 400um need to be designed by opening blastocyst stage.
Preferably, the thickness of Embryo Culture position limiting fence separation lattice can be adjusted according to the intensity of material in the step S2, and The thickness of Embryo Culture position limiting fence edge frame can be adjusted according to the intensity of material.
Preferably, the four walls punching of each culture lattice of Embryo Culture position limiting fence can be according to material in the step S4 Intensity and the adjustment of embryo's size.
(3) beneficial effect
The present invention provides a kind of Embryo Culture position limiting fences.Have compared with prior art following the utility model has the advantages that the embryo Position limiting fence is cultivated, by the production method in Embryo Culture position limiting fence specifically includes the following steps: S1, Embryo Culture position limiting fence Material selection, chooses polystyrene or other medical materials carry out the production of Embryo Culture position limiting fence, S2, Embryo Culture position limiting fence Size design, according to embryo's diameter within the scope of 180-220um, can design each separation the long 300um of lattice inside dimension, it is wide Latticed Embryo Culture limit can be obtained in 300um, high 200um, each separation lattice thickness 50um, four frame thickness 0.6mm of edge Column, the logo design of S3, Embryo Culture position limiting fence mark English alphabet, mark at ordinate at each culture lattice abscissa Remember Arabic numerals, so each culture lattice has unique double figures to identify, the liquid flowing of S4, Embryo Culture position limiting fence Property design, each culture lattice four walls, punched using laser or plasma technology, each bore dia be 50um, hole Quantity and size can be adjusted according to the size of embryo, and S5, manufactured Embryo Culture position limiting fence will be surface-treated, and embryo is trained Support position limiting fence makes the surface of Embryo Culture position limiting fence have hydrophilic or hydrophobicity, S6, manufactured embryo by process of surface treatment Culture position limiting fence will just can be used by ethylene oxide sterilizing or radiosterilization, and Embryo Culture position limiting fence surface is made to be in nothing Embryo Culture position limiting fence is put into the hole of four well culture plates by bacterium state, the use of S7, Embryo Culture position limiting fence, adjusts position The center for occuping hole is set, S8, then 500ul culture solution is injected into the hole of four well culture plates, pays attention to not making Embryo Culture Position limiting fence floats, if having air in the four walls cavity of culture lattice, can use micropipettor pressure-vaccum repeatedly, be removed, S9, general Above culture solution in the careful hole for being injected into four well culture plates of 300ul mineral oil, S10, the cumulus oocyte by acquirement Compound either fertilized embryo, is cleaned in the culture lattice for be put into three times Embryo Culture position limiting fence with culture solution and is cultivated, can It realizes and makes good Embryo Culture position limiting fence effectively to overcome single embryo in vitro in incubation the phenomenon that depauperation, Simplify operating procedure simultaneously, has achieved the purpose that ensure to distinguish embryonic origin well, to greatly facilitate the embryo of people Training.
Detailed description of the invention
Fig. 1 is the plane partial structural diagram of Embryo Culture position limiting fence of the present invention;
Fig. 2 is Embryo Culture position limiting fence 3D partial structural diagram of the present invention.
Specific embodiment
Following will be combined with the drawings in the embodiments of the present invention, and technical solution in the embodiment of the present invention carries out clear, complete Site preparation description, it is clear that described embodiments are only a part of the embodiments of the present invention, instead of all the embodiments.It is based on Embodiment in the present invention, it is obtained by those of ordinary skill in the art without making creative efforts every other Embodiment shall fall within the protection scope of the present invention.
The embodiment of the present invention provides a kind of technical solution referring to FIG. 1-2: a kind of Embryo Culture position limiting fence, Embryo Culture The production method of position limiting fence specifically includes the following steps:
The material selection of S1, Embryo Culture position limiting fence, choose polystyrene or other medical materials carry out Embryo Culture limit The production of bit field;
The size design of S2, Embryo Culture position limiting fence can design each according to embryo's diameter within the scope of 180-220um A separation lattice inside dimension long 300um, wide 300um, high 200um, each separation lattice thickness 50um, four frame thickness of edge Latticed Embryo Culture position limiting fence can be obtained in 0.6mm;
The logo design of S3, Embryo Culture position limiting fence mark English alphabet, indulge and sit at each culture lattice abscissa Arabic numerals are marked at mark, so each culture lattice has unique double figures to identify;
The liquid fluidity design of S4, Embryo Culture position limiting fence, four walls of each culture lattice, using laser or plasma Technology is punched, and each bore dia is 50um, and the quantity and size in hole can be adjusted according to the size of embryo;
S5, manufactured Embryo Culture position limiting fence will be surface-treated, and Embryo Culture position limiting fence is passed through process of surface treatment Making the surface of Embryo Culture position limiting fence has hydrophilic or hydrophobicity;
S6, manufactured Embryo Culture position limiting fence will just can be used by ethylene oxide sterilizing or radiosterilization, make embryo Tire culture position limiting fence surface is in germ-free condition;
Embryo Culture position limiting fence is put into the hole of four well culture plates by the use of S7, Embryo Culture position limiting fence, adjusts position Set the center for occuping hole;
S8, then 500ul culture solution is injected into the hole of four well culture plates, pays attention to not making Embryo Culture position limiting fence floating It rises, if having air in the four walls cavity of culture lattice, micropipettor pressure-vaccum repeatedly can be used, be removed;
S9, will be above the culture solution in the careful hole for being injected into four well culture plates of 300ul mineral oil;
S10, by the cumulus oocyte compound of acquirement either fertilized embryo, cleaned three times with culture solution, be put into embryo It is cultivated in the culture lattice of tire culture position limiting fence.
In the present invention, the size of Embryo Culture position limiting fence can be adjusted according to the stage of development of embryo in step S2, expanded Blastocyst stage need to design the long 600um of Embryo Culture position limiting fence, wide 600um, high 400um.
In the present invention, the thickness of Embryo Culture position limiting fence separation lattice can be adjusted according to the intensity of material in step S2, and embryo The thickness of tire culture position limiting fence edge frame can be adjusted according to the intensity of material.
In the present invention, the four walls punching of each culture lattice of Embryo Culture position limiting fence can be according to the strong of material in step S4 Degree and the adjustment of embryo's size.
It should be noted that, in this document, relational terms such as first and second and the like are used merely to a reality Body or operation are distinguished with another entity or operation, are deposited without necessarily requiring or implying between these entities or operation In any actual relationship or order or sequence.Moreover, the terms "include", "comprise" or its any other variant are intended to Non-exclusive inclusion, so that the process, method, article or equipment including a series of elements is not only wanted including those Element, but also including other elements that are not explicitly listed, or further include for this process, method, article or equipment Intrinsic element.
It although an embodiment of the present invention has been shown and described, for the ordinary skill in the art, can be with A variety of variations, modification, replacement can be carried out to these embodiments without departing from the principles and spirit of the present invention by understanding And modification, the scope of the present invention is defined by the appended.

Claims (4)

1. a kind of Embryo Culture position limiting fence, it is characterised in that: the production method of the Embryo Culture position limiting fence specifically includes following Step:
The material selection of S1, Embryo Culture position limiting fence, choose polystyrene or other medical materials carry out Embryo Culture position limiting fence Production;
The size design of S2, Embryo Culture position limiting fence can design each point according to embryo's diameter within the scope of 180-220um The long 300um of inside dimension, wide 300um, high 200um out of place, each separation lattice thickness 50um, four frame thickness 0.6mm of edge, can Obtain latticed Embryo Culture position limiting fence;
The logo design of S3, Embryo Culture position limiting fence mark English alphabet, at ordinate at each culture lattice abscissa Arabic numerals are marked, so each culture lattice has unique double figures to identify;
The liquid fluidity design of S4, Embryo Culture position limiting fence, four walls of each culture lattice, using laser or plasma technology It is punched, each bore dia is 50um, and the quantity and size in hole can be adjusted according to the size of embryo;
S5, manufactured Embryo Culture position limiting fence will be surface-treated, and Embryo Culture position limiting fence is made embryo by process of surface treatment The surface of tire culture position limiting fence has hydrophilic or hydrophobicity;
S6, manufactured Embryo Culture position limiting fence will just can be used by ethylene oxide sterilizing or radiosterilization, train embryo It supports position limiting fence surface and is in germ-free condition;
Embryo Culture position limiting fence is put into the hole of four well culture plates by the use of S7, Embryo Culture position limiting fence, and adjustment position occupies In the center in hole;
S8, then 500ul culture solution is injected into the hole of four well culture plates, pays attention to not floating Embryo Culture position limiting fence, If having air in the four walls cavity of culture lattice, micropipettor pressure-vaccum repeatedly can be used, be removed;
S9, will be above the culture solution in the careful hole for being injected into four well culture plates of 300ul mineral oil;
S10, by the cumulus oocyte compound of acquirement either fertilized embryo, clean three times with culture solution, be put into embryo train It supports and is cultivated in the culture lattice of position limiting fence.
2. a kind of Embryo Culture position limiting fence according to claim 1, it is characterised in that: Embryo Culture limits in the step S2 The size of bit field can be adjusted according to the stage of development of embryo, need to design the long 600um of Embryo Culture position limiting fence in the Blastocysts stage, Wide 600um, high 400um.
3. a kind of Embryo Culture position limiting fence according to claim 1, it is characterised in that: Embryo Culture limits in the step S2 The thickness of bit field separation lattice can be adjusted according to the intensity of material, and the thickness of Embryo Culture position limiting fence edge frame can be according to material Intensity adjustment.
4. a kind of Embryo Culture position limiting fence according to claim 1, it is characterised in that: Embryo Culture limits in the step S4 The four walls punching of each culture lattice of bit field can be adjusted according to the intensity and embryo's size of material.
CN201811513114.0A 2018-12-11 2018-12-11 A kind of Embryo Culture position limiting fence Pending CN109486677A (en)

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Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5578490A (en) * 1992-09-16 1996-11-26 Universidade De Santiago De Compostela Cell culture plate with a system for lateral diffusion of molecules through a barrier membrane
US6448069B1 (en) * 2000-06-09 2002-09-10 Genx International Corp. Embryo-culturing apparatus and method
US20080003672A1 (en) * 2006-06-29 2008-01-03 Cecchi Michael D Apparatus and method for culturing oocytes, embryos, stem cells and cells
CN202945247U (en) * 2012-12-07 2013-05-22 广西壮族自治区妇幼保健院 Human embryo culture vessel with single culture, co-culture and group culture advantages
CN205258488U (en) * 2015-12-24 2016-05-25 梁晓东 Novel embryo culture ware
CN206418141U (en) * 2017-01-23 2017-08-18 中南大学 A kind of Embryo Culture ware
CN107058455A (en) * 2016-08-10 2017-08-18 黑龙江省农业科学院畜牧研究所 The measuring method of the cell stage volume of pig 2

Patent Citations (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5578490A (en) * 1992-09-16 1996-11-26 Universidade De Santiago De Compostela Cell culture plate with a system for lateral diffusion of molecules through a barrier membrane
US6448069B1 (en) * 2000-06-09 2002-09-10 Genx International Corp. Embryo-culturing apparatus and method
US20080003672A1 (en) * 2006-06-29 2008-01-03 Cecchi Michael D Apparatus and method for culturing oocytes, embryos, stem cells and cells
CN202945247U (en) * 2012-12-07 2013-05-22 广西壮族自治区妇幼保健院 Human embryo culture vessel with single culture, co-culture and group culture advantages
CN205258488U (en) * 2015-12-24 2016-05-25 梁晓东 Novel embryo culture ware
CN107058455A (en) * 2016-08-10 2017-08-18 黑龙江省农业科学院畜牧研究所 The measuring method of the cell stage volume of pig 2
CN206418141U (en) * 2017-01-23 2017-08-18 中南大学 A kind of Embryo Culture ware

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
李卓琦德国: "胚胎单个定位及三维共培养微流控装置的构建", 《清华大学学报》 *

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