CN105838607A - Cell cultivation insertion component for scratch experiments and application of cell cultivation insertion component - Google Patents
Cell cultivation insertion component for scratch experiments and application of cell cultivation insertion component Download PDFInfo
- Publication number
- CN105838607A CN105838607A CN201610364871.0A CN201610364871A CN105838607A CN 105838607 A CN105838607 A CN 105838607A CN 201610364871 A CN201610364871 A CN 201610364871A CN 105838607 A CN105838607 A CN 105838607A
- Authority
- CN
- China
- Prior art keywords
- cell
- cut
- chamber
- cell culture
- culture insert
- Prior art date
- 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.)
- Pending
Links
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
- C12M23/00—Constructional details, e.g. recesses, hinges
- C12M23/02—Form or structure of the vessel
- C12M23/10—Petri dish
-
- 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
-
- 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
- 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
- C12M41/46—Means for regulation, monitoring, measurement or control, e.g. flow regulation of cellular or enzymatic activity or functionality, e.g. cell viability
-
- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N33/00—Investigating or analysing materials by specific methods not covered by groups G01N1/00 - G01N31/00
- G01N33/48—Biological material, e.g. blood, urine; Haemocytometers
- G01N33/50—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing
- G01N33/5005—Chemical analysis of biological material, e.g. blood, urine; Testing involving biospecific ligand binding methods; Immunological testing involving human or animal cells
Abstract
The invention discloses a device for scratch experiments. The device comprises a cultivation dish or a cultivation plate or a glass slide with chambers and a cell cultivation insertion component. The cultivation dish or the cultivation plate or the glass slide with the chambers and the cell cultivation insertion component are used for cultivating cells. The cell cultivation insertion component is of an 'H'-shaped structure and is provided with at least one scratch barrier and four support barriers, the scratch barriers are used for forming scratch blank zones, the four support barriers are used for supporting the cell cultivation insertion component, clinging contact surfaces are formed by the bottom surface of the cell cultivation insertion component and cultivation surfaces of the cultivation dish or cultivation surfaces of the chambers, and the scratch blank zones can be formed in the scratch experiments and correspond to clinging contact surfaces of the scratch barriers. The device has the advantages that the neat scratch blank zones can be easily and conveniently formed in the scratch experiments by the cell cultivation insertion component in a special design, accordingly, operation steps of the scratch experiments can be greatly simplified, and the operability and the repeatability of the scratch experiments can be greatly enhanced; the cell cultivation insertion component, the cultivation device with the cell cultivation insertion component and application of the cell cultivation insertion component are implemented on the basis.
Description
Technical field
The present invention relates to cell cultivate and cell function analysis field, thin more particularly to for scratch experiment
Born of the same parents cultivate plug-in unit, culture apparatus containing described plug-in unit and application thereof.
Background technology
Scratch experiment is also wound healing assay, is used to analyze the experiment that cell migration, motor capacity are the most frequently used
Method.It is usually used to study movement cell such as podocyte etc., and the transfer ability of various tumor cell.
The traditional method doing scratch experiment is after cell confluent cultures ware, draws with rifle head, wipes cell, shape off
Become blank region.But, there is obvious shortcoming in the method, specifically includes that (1) scoring operation relies on operation
The experience of person；(2) clear area that cut is formed is the most neat, it is impossible to carry out determining quantitative analysis；(3) repeatability
Difference, the diversity (variation) of operation is bigger every time；(4) draw with rifle head, the bag bottom culture dish can be damaged
By thing, even destroy the flatness of lower surface so that some adherent cells can not grow in clear area, causes
The negative findings that cell migration ability is weak.
Due to existing cut method, to there is many deficiencies, such as data the most beautiful, scored area zigzag,
Irregular, poor repeatability, it is not easy to obtain quantitative result, more can not obtain statistical discrepancy, therefore, a lot of scientific researches
Worker, when doing related experiment, is typically only capable to do preliminary experiment with scratch experiment, or can only be by scratch experiment result
It is placed in supplementary data.
Therefore, this area is in the urgent need to developing height easy and simple to handle, repeated, even can be used for quantitative analysis and draw
The apparatus and method of trace experiment.
Summary of the invention
It is an object of the invention to provide a kind of height easy and simple to handle, repeated, even can be used for quantitative analysis cut
The apparatus and method of experiment.
In a first aspect of the present invention, it is provided that a kind of device for scratch experiment, described device includes:
(a) for cultivating the culture dish of cell or culture plate or chamber slide, wherein said culture plate with
And chamber slide has one or more chamber for cultivating cell；
B () cell culture insert, described cell culture insert has dies, and has flat
Smooth bottom surface, wherein said cell culture insert is provided with at least one and keeps off for the cut forming cut clear area
Plate and for supporting four supporting baffle of described cell culture insert, wherein the first supporting baffle and second
Supporting baffle is fixed on one end of described cut baffle plate, and the 3rd supporting baffle and the 4th supporting baffle are fixed on
The other end of described cut baffle plate, thus form " work " type structure；
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d；
Further, when described cell culture insert is placed in described culture dish or is placed in described chamber, described
The bottom surface of cell culture insert forms laminating with the cultivation face cultivating face or described chamber of described culture dish
Contact surface, and the laminating contact surface corresponding to described cut baffle plate forms cut blank in scratch experiment
District.
Preferably, described l is 0.5-10cm, and described d is 100-1000 μm.
In another preference, the angle a between described first supporting baffle and the second supporting baffle and institute
The angle b stated between the 3rd supporting baffle and the 4th supporting baffle is 30-330 degree.
In another preference, the absorption affinity between described cell culture insert and described culture dish is more than institute
State the gravity of cell culture insert.
In another preference, the cross section of described each supporting baffle is long strip type, arc or rectangle.
In another preference, the area Sz of the laminating contact surface that described each supporting baffle and the face of cultivation are formed
For 0.5-100mm2。
In another preference, described each supporting baffle is identical or different.
In another preference, the cross section of described one or more supporting baffle is rectangle, and corresponds to
The laminating contact surface of described one or more supporting baffle forms auxiliary cut clear area in scratch experiment.
In another preference, described culture dish is a diameter of 3.5cm, 6cm, 7cm, 9cm, 10cm, 15cm
Culture dish Deng different culture surface sizes.
In another preference, described chamber slide has 2,3,4,6,8,10,12 for training
Support the chamber of cell.
In another preference, described culture plate has 2,4,6,8,16,24,48 and 96 use
In the chamber cultivating cell.
In another preference, described cell culture insert is the silica gel plug-in unit of bio-compatibility.
In another preference, described cell culture insert has rough surface at an upper portion thereof.
In another preference, described cell culture insert has smooth planar surface in its underpart.
In a second aspect of the present invention, it is provided that a kind of consumptive material for scratch experiment is set with, described suit bag
Include:
(a) first container, and it is positioned at the culture dish of described first container or culture plate or chamber carries glass
Sheet, wherein said culture plate and described chamber slide have one or more chamber for cultivating cell
Room；
(b) second container, and it is positioned at the cell culture insert of described second container, described cell is trained
Foster plug-in unit has dies, and has smooth bottom surface, and wherein said cell culture insert sets
There is at least one for forming the cut baffle plate of cut clear area and for supporting described cell culture insert
Four supporting baffle, wherein the first supporting baffle and the second supporting baffle are fixed on the one of described cut baffle plate
End, and the 3rd supporting baffle and the 4th supporting baffle are fixed on the other end of described cut baffle plate, thus formed
" work " type structure；
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d, and described l is 0.5
-10cm, described d are 100-1000 μm；
Further, when described cell culture insert is placed in described culture dish or is placed in described chamber, described
The bottom surface of cell culture insert forms laminating with the cultivation face cultivating face or described chamber of described culture dish
Contact surface, and the laminating contact surface corresponding to described cut baffle plate forms cut blank in scratch experiment
District；
C operation instructions that () is optional.
In a third aspect of the present invention, it is provided that a kind of consumptive material for scratch experiment is set with, described suit bag
Include:
(a) first container, and it is positioned at the culture dish of described first container or culture plate or chamber carries glass
Sheet, wherein said culture plate and described chamber slide have one or more chamber for cultivating cell
Room, and in the chamber of each cultivation cell, preset cell culture insert, described cell culture insert has
Having dies, and have smooth bottom surface, wherein said cell culture insert is provided with at least one
Individual cut baffle plate for forming cut clear area and for supporting four of described cell culture insert
Spacer plate, wherein the first supporting baffle and the second supporting baffle are fixed on one end of described cut baffle plate, and
Three supporting baffle and the 4th supporting baffle are fixed on the other end of described cut baffle plate, thus form " work " type
Structure；
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d, and described l is 0.5
-10cm, described d are 100-1000 μm；
Further, when described cell culture insert is placed in described culture dish or is placed in described chamber, described
The bottom surface of cell culture insert forms laminating with the cultivation face cultivating face or described chamber of described culture dish
Contact surface, and the laminating contact surface corresponding to described cut baffle plate forms cut blank in scratch experiment
District；
B operation instructions that () is optional.
Preferably, described consumptive material is set with possibly together with second container, and is positioned at the thin of described second container
Born of the same parents cultivate the cell preset in plug-in unit, wherein said cell culture insert and described first container and cultivate slotting
The specification of part is same or different, and described specification is by the size and shape of the sectional area of described cut baffle plate
Divide.
In a fourth aspect of the present invention, it is provided that the mensuration cell migration energy of a kind of nondiagnostic and non-therapeutic
The method of power, including step:
A () provides the device described in first aspect, wherein said cell culture insert is placed on described training
Support in the chamber of ware or the chamber of described culture plate or described chamber slide so that described cell culture insert
Fit in cultivation face or described culture plate, the cultivation face of described chamber slide of described culture dish；
B () adds appropriate cell suspension in described device；
(c) cultivate described in cell, until described cell attachment and be paved with or be substantially paved with bottom cultivation
Face；
D () removes described cell culture insert, thus form cut clear area；
E () continues the cell described in cultivation, and observe and analyze the transfer ability of described cell.
Preferably, in step (d), after removing described cell culture insert, form smooth cut blank
District；
In step (e), by the described cell culture insert of same size in the described chamber of formed objects
The each cut clear area formed, carries out qualitative and/or quantitative and/or statistical analysis.
In another preference, in step (d), after removing described cell culture insert, form smooth drawing
Trace clear area, does not destroy the chamber of described culture dish or the chamber of described culture plate or described chamber slide
Bottom culture surface, cell can be at described culture surface adherent growth, travel motion.
In another preference, described cultivation face is slide surface.
In another preference, in step (b), the addition of described cell suspension is 50-15000 microlitre
/ chamber, according to culture dish used, culture plate, chamber slide concrete chamber size depending on.
In another preference, in step (b), the concentration of the middle cell of described cell suspension is 1 × 104
-1 × 106Individual cell/ml, depending on the cell type that concrete foundation is cultivated.
In another preference, the width d of described cut clear area is 100-1000 μm, and length l is
0.5-10cm.
In another preference, described culture plate includes the chamber of n identical or different size.
In another preference, described chamber slide includes the chamber of n identical or different size.
In another preference, in step (e), described analysis is quantitative analysis.
In another preference, in step (e), in formed objects chamber with same size described carefully
Born of the same parents cultivate each cut clear area that plug-in unit is formed, and carry out quantitatively and/or statistical analysis.
In should be understood that within the scope of the present invention, above-mentioned each technical characteristic of the present invention and below (such as embodiment)
Can be combined with each other between each technical characteristic of middle specific descriptions, thus constitute new or preferred technical scheme.
As space is limited, the most tired at this state.
Accompanying drawing explanation
Fig. 1 is the structural representation of scuffing experimental device in embodiment 1；
The structural representation of the cut clear area formed when Fig. 2 is scratch experiment in embodiment 1；
Fig. 3 is the structural representation of scuffing experimental device in embodiment 2；
Fig. 4 is the structural representation of scuffing experimental device in embodiment 3.
Detailed description of the invention
The present inventor, through extensively in-depth study, develops a kind of cell for scratch experiment first and trains
Support plug-in unit.By the biocompatible silica gel plug-in unit of this particular design, can simplicity be formed whole in scratch experiment
Neat cut clear area, thus enormously simplify the operating procedure of scratch experiment, greatly improve this experiment
Ease for operation and repeatability.Complete the present invention on this basis.
Term
As used herein, term " cell culture insert of the present invention ", " present invention cultivates plug-in unit ", " this
Invention plug-in unit " it is used interchangeably, refer to the cell culture insert of the present invention
As used herein, term " I shape " " H type " is used interchangeably, and phalangeal cell cultivates the shape of plug-in unit,
Supporting baffle can be linear type, shaped form, broken line type or a combination thereof, and such as, two baffle plates are (with first gear
As a example by plate and second baffle), when it is all linear type, a straight line (i.e. angle α=180 °) can be collectively formed,
Can also be constituted other arbitrarily angled, preferably 30-320 degree, 60-300 degree, 90-270 degree, 120-240 degree,
150-210 degree, thus constitute "<", ">" (for H type), " V " or inverted " V " (for " work "
For type) or other similar angle, such as " U ", " C " etc..
Scuffing experimental device
In one embodiment, it is provided that a kind of device for scratch experiment, described device includes:
(a) for cultivating the culture dish of cell or culture plate or chamber slide, wherein said culture plate with
And chamber slide has one or more chamber for cultivating cell；
B () cell culture insert, described cell culture insert has dies, and has flat
Smooth bottom surface, wherein said cell culture insert is provided with at least one and keeps off for the cut forming cut clear area
Plate and for supporting four supporting baffle of described cell culture insert, wherein the first supporting baffle and second
Supporting baffle is fixed on one end of described cut baffle plate, and the 3rd supporting baffle and the 4th supporting baffle are fixed on
The other end of described cut baffle plate, thus form " work " type structure.
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d.
Further, when described cell culture insert is placed in described culture dish or is placed in described chamber, described
The bottom surface of cell culture insert forms laminating with the cultivation face cultivating face or described chamber of described culture dish
Contact surface, and the laminating contact surface corresponding to described cut baffle plate forms cut blank in scratch experiment
District.
Preferably, described l is 0.5-10cm, and described d is 100-1000 μm.
In another preference, the angle a between described first supporting baffle and the second supporting baffle and institute
The angle b stated between the 3rd supporting baffle and the 4th supporting baffle is 30-330 degree.
In another preference, the absorption affinity between described cell culture insert and described culture dish is more than institute
State the gravity of cell culture insert.
In another preference, the cross section of described each supporting baffle is long strip type, arc or rectangle.
In another preference, the area Sz of the laminating contact surface that described each supporting baffle and the face of cultivation are formed
For 0.5-100mm2。
In another preference, described each supporting baffle is identical or different.
In another preference, the cross section of described one or more supporting baffle is rectangle, and corresponds to
The laminating contact surface of described one or more supporting baffle forms auxiliary cut clear area in scratch experiment.
In another preference, described culture dish is a diameter of 3.5cm, 6cm, 7cm, 9cm, 10cm, 15cm
Culture dish Deng different culture surface sizes.
In another preference, described chamber slide has 2,3,4,6,8,10,12 for training
Support the chamber of cell.
In another preference, described culture plate has 2,4,6,8,16,24,48 and 96 use
In the chamber cultivating cell.
In another preference, described cell culture insert is the silica gel plug-in unit of bio-compatibility.
In another preference, described cell culture insert has rough surface at an upper portion thereof.
In another preference, described cell culture insert has smooth planar surface in its underpart.
In another preference, described silica gel plug-in unit can make it be attached to chamber slide by electrostatic interaction
Bottom, the bottom fixed width of silica gel plug-in unit is 500 μm.
In another preference, the bottom (contact surface i.e. contacted with chamber slide surface) of described silica gel plug-in unit
It is flat, smooth, thus is possible not only to form good laminating with chamber slide surface and contacts, thus suction is provided
Attached power (suction), additionally, described silica gel plug-in unit can make it solid by active forces such as electrostatic interactions further
Determine or be adsorbed in chamber slide.
In another preference, the upper and lower of described silica gel plug-in unit is constructed from the same material.
In another preference, the upper and lower of described silica gel plug-in unit is made up of different materials.Such as,
It is coarse material that top uses, and bottom uses finer and smoother material.
In another preference, although the upper and lower of described silica gel plug-in unit is by identical or essentially identical material
Material is constituted, but top has the most coarse surface, and bottom has smooth surface.So, it is simple to plug-in unit
It is firmly fixed to culture dish, culture plate, the cultivation face of chamber slide；It addition, when carrying out removing operation,
Easily remove plug-in unit of the present invention, impact without cell migration experiment.
Consumptive material is set with
In another embodiment, it is provided that a kind of consumptive material for scratch experiment is set with, and described suit includes:
(a) first container, and it is positioned at the culture dish of described first container or culture plate or chamber carries glass
Sheet, wherein said culture plate and described chamber slide have one or more chamber for cultivating cell
Room；
(b) second container, and it is positioned at the cell culture insert of described second container, described cell is trained
Foster plug-in unit has dies, and has smooth bottom surface, and wherein said cell culture insert sets
There is at least one for forming the cut baffle plate of cut clear area and for supporting described cell culture insert
Four supporting baffle, wherein the first supporting baffle and the second supporting baffle are fixed on the one of described cut baffle plate
End, and the 3rd supporting baffle and the 4th supporting baffle are fixed on the other end of described cut baffle plate, thus formed
" work " type structure；
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d, and described l is 0.5
-10cm, described d are 100-1000 μm；
Further, when described cell culture insert is placed in described culture dish or is placed in described chamber, described
The bottom surface of cell culture insert forms laminating with the cultivation face cultivating face or described chamber of described culture dish
Contact surface, and the laminating contact surface corresponding to described cut baffle plate forms cut blank in scratch experiment
District；
C operation instructions that () is optional.
Consumptive material is set with
In another embodiment, it is provided that a kind of consumptive material for scratch experiment is set with, and described suit includes:
(a) first container, and it is positioned at the culture dish of described first container or culture plate or chamber carries glass
Sheet, wherein said culture plate and described chamber slide have one or more chamber for cultivating cell
Room, and in the chamber of each cultivation cell, preset cell culture insert, described cell culture insert has
Having dies, and have smooth bottom surface, wherein said cell culture insert is provided with at least one
Individual cut baffle plate for forming cut clear area and for supporting four of described cell culture insert
Spacer plate, wherein the first supporting baffle and the second supporting baffle are fixed on one end of described cut baffle plate, and
Three supporting baffle and the 4th supporting baffle are fixed on the other end of described cut baffle plate, thus form " work " type
Structure；
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d, and described l is 0.5
-10cm, described d are 100-1000 μm；
Further, when described cell culture insert is placed in described culture dish or is placed in described chamber, described
The bottom surface of cell culture insert forms laminating with the cultivation face cultivating face or described chamber of described culture dish
Contact surface, and the laminating contact surface corresponding to described cut baffle plate forms cut blank in scratch experiment
District；
B operation instructions that () is optional.
Preferably, described consumptive material is set with possibly together with second container, and is positioned at the thin of described second container
Born of the same parents cultivate the cell preset in plug-in unit, wherein said cell culture insert and described first container and cultivate slotting
The specification of part is same or different, and described specification is by the size and shape of the sectional area of described cut baffle plate
Divide.
The method measuring cell migration ability
In another embodiment, it is provided that the mensuration cell migration ability of a kind of nondiagnostic and non-therapeutic
Method, including step:
A () provides the device described in scuffing experimental device, wherein said cell culture insert is placed on institute
State in the chamber of culture dish or the chamber of described culture plate or described chamber slide so that described cell is cultivated
Plug-in unit fits in cultivation face or described culture plate, the cultivation face of described chamber slide of described culture dish；
B () adds appropriate cell suspension in described device；
(c) cultivate described in cell, until described cell attachment and be paved with or be substantially paved with bottom cultivation
Face；
D () removes described cell culture insert, thus form cut clear area；
E () continues the cell described in cultivation, and observe and analyze the transfer ability of described cell.
Preferably, in step (d), after removing described cell culture insert, form smooth cut blank
District；
In step (e), by the described cell culture insert of same size in the described chamber of formed objects
The each cut clear area formed, carries out qualitative and/or quantitative and/or statistical analysis.
In another preference, in step (d), after removing described cell culture insert, form smooth drawing
Trace clear area, does not destroy the chamber of described culture dish or the chamber of described culture plate or described chamber slide
Bottom culture surface, cell can be at described culture surface adherent growth, travel motion.
In another preference, described cultivation face is slide surface.
In another preference, in step (b), the addition of described cell suspension be 50-5000 milliliter/
Chamber, according to culture dish used, culture plate, chamber slide concrete chamber size depending on.
In another preference, in step (b), the concentration of the middle cell of described cell suspension is 1 × 104
-1 × 106Individual cell/ml, depending on the cell type that concrete foundation is cultivated.
In another preference, the width d of described cut clear area is 100-1000 μm, and length l is
0.5-10cm.
In another preference, described culture plate includes the chamber of n identical or different size.
In another preference, described chamber slide includes the chamber of n identical or different size.
In another preference, in step (e), described analysis is quantitative analysis.
In another preference, in step (e), in formed objects chamber with same size described carefully
Born of the same parents cultivate each cut clear area that plug-in unit is formed, and carry out quantitatively and/or statistical analysis.
In the present embodiment, for the cut clear area of two plug-in units formation of same specification, it differs amplitude
Can be calculated as follows:
Difference amplitude F (%)=100% × | d1-d2 |/((d1+d2)/2)
In formula, d1 is the width (or span) of first cut clear area；
D2 is the width (or span) of second cut clear area.
The cut clear area that n plug-in unit of same specification is formed, its difference amplitude can be calculated as follows:
Difference amplitude F (%)=100% × | dmax-dmin|/daverage
In formula,
dmaxWidth (or span) for the cut clear area of width maximum；
dminWidth (or span) for the cut clear area of width minimum；
daverageGenerally, mean breadth (or average span) (d for n cut clear areaaverageWith same specification plug-in unit
Design width identical or essentially identical).
Material and universal method
Cell: Human umbilical vein endothelial cells (HUVEC).
Culture medium: Endothelial cell culture base, 10% hyclone.
Cultural method: 37 DEG C, 5%CO2, cultivate in cell culture incubator.
Cell migration assay: microscope obtains experimental image, ImageJ software or WimScratch software
The area coverage in analysis margin region.
It should be noted that in the claim and description of this patent, such as first and second or the like
Relational terms is used merely to separate an entity or operation with another entity or operating space, and is not necessarily intended to
Seek or imply relation or the order that there is any this reality between these entities or operation.And, term " bag
Include ", " comprising " or its any other variant be intended to comprising of nonexcludability, so that include that one is
The process of row key element, method, article or equipment not only include those key elements, but also include being not expressly set out
Other key elements, or also include the key element intrinsic for this process, method, article or equipment.Do not having
In the case of having more restriction, statement the key element " including " and limiting, it is not excluded that including described key element
Process, method, article or equipment in there is also other identical element.
Main advantages of the present invention include:
A () cell culture insert of the present invention rational in infrastructure, is highly suitable for scratch experiment.
When () carries out scratch experiment by cell culture insert of the present invention b, simple to operate, the saving time.
C () present invention can form width certain (such as 200 microns, 300 microns, 500 microns etc.), region
Significantly clear area (being i.e. similar to the cut district of routine), it is easy to carry out quantitative analysis.
D scratch experiment that () is carried out by cell culture insert of the present invention, repeatability is the highest.
E the cell culture insert of () present invention is based on excellent fastness: in drop test, and plug-in unit is solid
Tissue Culture Dish after Ding is inverted, and plug-in unit of the present invention will not fall；Additionally, after addition cell suspension, having
In the case of solution, after cultivating 48 hours (meeting the duration needed for scratch experiment), leakage will not occur.
F () cell culture insert of the present invention is fixed in Tissue Culture Dish, culture plate or chamber slide after,
After outer package processes, can transport for long-distance, keep gnotobasis, plug-in unit does not comes off.
Below in conjunction with specific embodiment, the present invention is expanded on further.Should be understood that these embodiments are only used for
The bright present invention rather than restriction the scope of the present invention.The experiment side of unreceipted actual conditions in the following example
Method, generally according to normal condition, or according to the condition proposed by manufacturer.Unless otherwise indicated, otherwise
Percentage ratio and number are percentage by weight and parts by weight.
Embodiment 1
Culture dish is used to carry out scratch experiment
Fig. 1 is the structural representation of scuffing experimental device in the present embodiment, and Fig. 2 is that in the present embodiment, cut is real
The structural representation of the cut clear area formed when testing.
As it is shown in figure 1, present embodiments provide a kind of device for scratch experiment, described device includes:
A () is for cultivating the culture dish 7 of cell；
B () cell culture insert 8, described cell culture insert 8 has dies, and has
Smooth bottom surface, wherein said cell culture insert 8 is had to be provided with one for forming drawing of cut clear area 9
Trace baffle plate 6 and for supporting four supporting baffle, wherein the first support blocks of described cell culture insert 8
Plate 1 and the second supporting baffle 2 are fixed on one end of described cut baffle plate 6, and the 3rd supporting baffle 3 and
Four supporting baffle 4 are fixed on the other end of described cut baffle plate 6, thus form " work " type structure.
Wherein, the cross section of described cut baffle plate 6 is length l, the rectangle of width d.
Further, in described cell culture insert 8 is placed in described culture dish 7, described cell culture insert
The bottom surface of 8 forms laminating contact surface with the cultivation face of described culture dish 7, and corresponding to described cut gear
The laminating contact surface of plate 6 forms cut clear area 9 in scratch experiment.
In the present embodiment, described l is 3cm, and described d is 500 μm.
Angle a between described first supporting baffle 1 and the second supporting baffle 2 and described 3rd support block
Angle b between plate 3 and the 4th supporting baffle 4 is 180 degree.
Described cell culture insert 8 makes its cultivation face being attached to culture dish 7, and institute by electrostatic interaction
Absorption affinity between cell culture insert 8 and the described culture dish 7 stated is more than described cell culture insert 8
Gravity.
Described each supporting baffle is identical, and the cross section of each supporting baffle is long strip type.
Described each supporting baffle with the area Sz of the laminating contact surface of the cultivation face formation of culture dish 7 is
10mm2。
The cross section of described supporting baffle is rectangle, and the laminating contact surface corresponding to described supporting baffle exists
Scratch experiment is formed auxiliary cut clear area 9.
Described culture dish 7 is the culture dish 7 of a diameter of 6cm culture surface size.
The described silica gel plug-in unit 8 that cell culture insert 8 is bio-compatibility, and the end of described silica gel plug-in unit 8
Portion's fixed width is 500 μm.
The upper and lower of described cell culture insert 8 is made up of different materials: it is coarse material that top uses
Material, and bottom uses finer and smoother material.
Device in embodiment 1 is tested as follows:
Tissue Culture Dish 7 a cell culture insert 8 is fixed by () after is inverted；
(b) in culture dish 7 disposable add needed for cell suspension, addition is 1000 microlitres,
The concentration of cell is 1 × 105Individual cell/ml, after cultivating 48 hours (meeting the duration needed for scratch experiment),
Remove cell culture insert 8；
C cell culture insert 8 is fixed on culture dish 7 by (), after outer package processes, transport for long-distance.
Test through related experiment personnel tracking, obtain following result:
A Tissue Culture Dish 7 is inverted by (), cell culture insert 8 will not be fallen；
B () cultivates 48 hours after, cell culture insert 8 will not occur leakage, removes cell and cultivates slotting
Part 8, cut clear area 9 is obvious, and cut clear area 9 width is unanimously 500 microns (as shown in Figure 2),
Cell growth state is good；
C, during () long-distance transport, cell culture insert 8 does not comes off.
Comparative example 1
Using the device (include material, specification) identical with embodiment 1, difference is: comparative example
Cell culture insert in 1 is " day " type structure；Cell culture insert in comparative example 1 is processed by glue
Fit with the cultivation face of culture dish.
Device in comparative example 1 is tested as follows:
(a ') cell culture insert is fixed on Tissue Culture Dish, then remove；
(b ') adds the cell suspension of pretreatment in culture dish, after cultivating 48 hours, removes cell
Cultivate plug-in unit, add cell culture fluid；
(c ') cell culture insert is fixed on culture dish, after outer package processes, transport for long-distance.
Test through related experiment personnel tracking, obtain following result:
(a ') processes the cultivation face that plug-in unit is fixed on culture dish with glue, is not easy to operation when removing plug-in unit,
The characteristics such as the hydrophilic in cultivation face, smoothness can be destroyed, be unfavorable for the growth of attached cell, and then easily make
Become the negative findings of cell migration ability；
After (b ') cultivates 48 hours, cell poor growth, remove cell culture insert, add cell
After culture fluid, the growth conditions of cell is bad；
During (c ') long-distance transport, cell culture insert can come off.
The above results shows: plug-in unit 8 structure of the I-shaped of the present invention is very beneficial for the growth of cell；
The plug-in unit 8 of the present invention can form that width is certain, obvious cut clear area, region 9, it is easy to quantitatively divides
Analysis；The cell culture insert 8 of the present invention is fixed on Tissue Culture Dish 7 by electrostatic interaction, will not during inversion
Falling, during long-distance transport, plug-in unit 8 does not comes off；The cell culture insert 8 of the present invention is fixed on cell training
Support ware 7, in the case of having solution, after cultivating 48 hours, leakage, and cell growth state will not occur
Well.
Embodiment 2
Culture dish is used to carry out scratch experiment
Fig. 3 is the structural representation of scuffing experimental device in the present embodiment.
As it is shown on figure 3, in the present embodiment, using device identical in embodiment 1, difference is:
Angle in the present embodiment, between the first supporting baffle 1 and second supporting baffle 2 of cell culture insert 8
Angle b between a and described 3rd supporting baffle 3 and the 4th supporting baffle 4 is 60 degree；Cut baffle plate
Cross-sectional length l of 6 is 4cm, and width d is 350 μm.
Cell is cultivated 6-8 hour, after cell is paved with cell cultivation face substantially, removes described plug-in unit 8.
Then, continuing under the same conditions to cultivate 6-10 hour, microscope obtains experimental image, and software divides
The area coverage of analysis cut clear area.
Result: after plug-in unit 8 removes, forms the cut clear area that obvious size (width) is 350 microns.
Embodiment 3
The microscope slide using 2 vestibule rooms carries out scratch experiment
Fig. 4 is the structural representation of scuffing experimental device in the present embodiment.
As shown in Figure 4, in the present embodiment, cell culture insert 8 identical in embodiment 1 is used, no
It is with point, uses the culture dish 7 in microscope slide 11 alternate embodiment 1 with 2 vestibule rooms 10, and
Placing " work " type plug-in unit 8 in each chamber 10 respectively, the size of " work " type plug-in unit 8 is with implementing
Example 1.
Cell is cultivated 6-8 hour, after cell is paved with cell cultivation face substantially, removes described " work " type
Plug-in unit 8.
Then, continuing under the same conditions to cultivate 6-10 hour, microscope obtains experimental image, and software divides
The area coverage of analysis cut clear area.
Result: after the plug-in unit 8 in 2 vestibule rooms 10 removes, being respectively formed obvious size (width) is 500
The cut clear area of micron, and width d1 and d2 difference amplitude≤1% of two cut clear areas.
Embodiment 4
The microscope slide using 4 vestibule rooms carries out scratch experiment
In the present embodiment, using cell culture insert identical in embodiment 1, difference is, uses
There is the culture dish in the microscope slide alternate embodiment 1 of 4 vestibule rooms, and in each chamber, place one respectively
Individual " work " type plug-in unit, the size of " work " type plug-in unit is with embodiment 1.
Cell is cultivated 6-8 hour, after cell is paved with cell cultivation face substantially, removes described " work " type
Plug-in unit.
Then, continuing under the same conditions to cultivate 6-10 hour, microscope obtains experimental image, and software divides
The area coverage of analysis cut clear area.
Result: after the plug-in unit in 4 vestibules removes, being respectively formed obvious size (width) is 500 microns
Difference amplitude≤1% between cut clear area, and width d1, d2, d3 and d4 of 4 cut clear areas.
Being put in by " work " type plug-in unit in the different chamber microscope slides such as 2 holes, 4 holes, 8 holes, benefit is as follows:
Save cell (cell concentration that culture dish needs is many, waste)；Conveniently do control experiment, repeat experiment；Same
On one microscope slide, it is easy to operation；Cost-effective；An equal amount of insert design makes to carry at different chamber
Slide is tested, when carrying out the experiment of different batches, not by the limitation and restriction of cultivation consumptive material, real
Test result and there is comparability.
Embodiment 5
Present embodiments providing a kind of consumptive material for scratch experiment to be set with, described suit includes:
(a) first container, and it is positioned at the culture dish of described first container；
(b) second container, and it is positioned at the cell culture insert of described second container, described cell is trained
Foster plug-in unit has dies, and has smooth bottom surface, and wherein said cell culture insert sets
There is one for forming the cut baffle plate of cut clear area and for supporting the four of described cell culture insert
Individual supporting baffle, wherein the first supporting baffle and the second supporting baffle are fixed on one end of described cut baffle plate,
And the 3rd supporting baffle and the 4th supporting baffle are fixed on the other end of described cut baffle plate, thus form " work "
Type structure.
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d, and described l is 3cm,
Described d is 500 μm.
Further, in described cell culture insert is placed in described culture dish, the end of described cell culture insert
Face forms laminating contact surface with the cultivation face of described culture dish, and corresponding to the laminating of described cut baffle plate
Contact surface forms cut clear area in scratch experiment.
Embodiment 6
Present embodiments providing a kind of consumptive material for scratch experiment to be set with, described suit includes:
(a) first container, and it is positioned at the culture plate of described first container, wherein said culture plate
There is multiple chamber for cultivating cell, and in the chamber of each cultivation cell, preset cell cultivation
Plug-in unit, described cell culture insert has dies, and has smooth bottom surface, Qi Zhongsuo
State cell culture insert and be provided with one for forming the cut baffle plate of cut clear area and described for supporting
Four supporting baffle of cell culture insert, wherein the first supporting baffle and the second supporting baffle are fixed on described
One end of cut baffle plate, and the 3rd supporting baffle and the 4th supporting baffle are fixed on another of described cut baffle plate
End, thus form " work " type structure；
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d, and described l is 8cm,
Described d is 800 μm；
Further, in described cell culture insert is placed in described chamber, the bottom surface of described cell culture insert
Form laminating contact surface with the cultivation face of described chamber, and the laminating corresponding to described cut baffle plate connects
Contacting surface forms cut clear area in scratch experiment；
(b) second container, and it is positioned at the cell culture insert of described second container, wherein said
The specification of the cell culture insert preset in cell culture insert and described first container is identical, described
Specification is divided by the size and shape of the sectional area of described cut baffle plate.
The all documents mentioned in the present invention are incorporated as reference the most in this application, just as each document
It is individually recited as with reference to like that.In addition, it is to be understood that after the above-mentioned teachings having read the present invention,
The present invention can be made various changes or modifications by those skilled in the art, and these equivalent form of values fall within this Shen equally
Please appended claims limited range.
Claims (10)
1. the device for scratch experiment, it is characterised in that described device includes:
(a) for cultivating the culture dish of cell or culture plate or chamber slide, wherein said culture plate with
And chamber slide has one or more chamber for cultivating cell；
B () cell culture insert, described cell culture insert has dies, and has flat
Smooth bottom surface, wherein said cell culture insert is provided with at least one and keeps off for the cut forming cut clear area
Plate and for supporting four supporting baffle of described cell culture insert, wherein the first supporting baffle and second
Supporting baffle is fixed on one end of described cut baffle plate, and the 3rd supporting baffle and the 4th supporting baffle are fixed on
The other end of described cut baffle plate, thus form " work " type structure；
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d；
Further, when described cell culture insert is placed in described culture dish or is placed in described chamber, described
The bottom surface of cell culture insert forms laminating with the cultivation face cultivating face or described chamber of described culture dish
Contact surface, and the laminating contact surface corresponding to described cut baffle plate forms cut blank in scratch experiment
District.
Device the most according to claim 1, it is characterised in that described l is 0.5-10cm, described
D is 100-1000 μm.
Device the most according to claim 1, it is characterised in that described first supporting baffle and second
The angle b between angle a and described 3rd supporting baffle and the 4th supporting baffle between supporting baffle is
30-330 degree.
Device the most according to claim 1, it is characterised in that described cell culture insert and institute
State the gravity more than described cell culture insert of the absorption affinity between culture dish.
5. the consumptive material suit for scratch experiment, it is characterised in that described suit includes:
(a) first container, and it is positioned at the culture dish of described first container or culture plate or chamber carries glass
Sheet, wherein said culture plate and described chamber slide have one or more chamber for cultivating cell
Room；
(b) second container, and it is positioned at the cell culture insert of described second container, described cell is trained
Foster plug-in unit has dies, and has smooth bottom surface, and wherein said cell culture insert sets
There is at least one for forming the cut baffle plate of cut clear area and for supporting described cell culture insert
Four supporting baffle, wherein the first supporting baffle and the second supporting baffle are fixed on the one of described cut baffle plate
End, and the 3rd supporting baffle and the 4th supporting baffle are fixed on the other end of described cut baffle plate, thus formed
" work " type structure；
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d, and described l is 0.5
-10cm, described d are 100-1000 μm；
Further, when described cell culture insert is placed in described culture dish or is placed in described chamber, described
The bottom surface of cell culture insert forms laminating with the cultivation face cultivating face or described chamber of described culture dish
Contact surface, and the laminating contact surface corresponding to described cut baffle plate forms cut blank in scratch experiment
District；
C operation instructions that () is optional.
6. the consumptive material suit for scratch experiment, it is characterised in that described suit includes:
(a) first container, and it is positioned at the culture dish of described first container or culture plate or chamber carries glass
Sheet, wherein said culture plate and described chamber slide have one or more chamber for cultivating cell
Room, and in the chamber of each cultivation cell, preset cell culture insert, described cell culture insert has
Having dies, and have smooth bottom surface, wherein said cell culture insert is provided with at least one
Individual cut baffle plate for forming cut clear area and for supporting four of described cell culture insert
Spacer plate, wherein the first supporting baffle and the second supporting baffle are fixed on one end of described cut baffle plate, and
Three supporting baffle and the 4th supporting baffle are fixed on the other end of described cut baffle plate, thus form " work " type
Structure；
Wherein, the cross section of described cut baffle plate is length l, the rectangle of width d, and described l is 0.5
-10cm, described d are 100-1000 μm；
Further, when described cell culture insert is placed in described culture dish or is placed in described chamber, described
The bottom surface of cell culture insert forms laminating with the cultivation face cultivating face or described chamber of described culture dish
Contact surface, and the laminating contact surface corresponding to described cut baffle plate forms cut blank in scratch experiment
District；
B operation instructions that () is optional.
Consumptive material the most according to claim 6 is set with, it is characterised in that described consumptive material suit also contains
Having a second container, and be positioned at the cell culture insert of described second container, wherein said cell is cultivated
The specification of the cell culture insert preset in plug-in unit and described first container is same or different, described
Specification is divided by the size and shape of the sectional area of described cut baffle plate.
8. a nondiagnostic and the method measuring cell migration ability of non-therapeutic, it is characterised in that
Including step:
A () provides the device described in claim 1, wherein said cell culture insert is placed on described
In the chamber of culture dish or the chamber of described culture plate or described chamber slide so that described cell is cultivated and inserted
Part fits in cultivation face or described culture plate, the cultivation face of described chamber slide of described culture dish；
B () adds appropriate cell suspension in described device；
(c) cultivate described in cell, until described cell attachment and be paved with or be substantially paved with bottom cultivation
Face；
D () removes described cell culture insert, thus form cut clear area；
E () continues the cell described in cultivation, and observe and analyze the transfer ability of described cell.
Method the most according to claim 8, it is characterised in that in step (d), remove described carefully
After born of the same parents cultivate plug-in unit, form smooth cut clear area；
In step (e), by the described cell culture insert of same size in the described chamber of formed objects
The each cut clear area formed, carries out qualitative and/or quantitative and/or statistical analysis.
Method the most according to claim 8, it is characterised in that in step (d), remove described carefully
After born of the same parents cultivate plug-in unit, form smooth cut clear area, do not destroy described culture dish or the chamber of described culture plate
The bottom culture surface of the chamber of room or described chamber slide, cell can be in the adherent life of described culture surface
Long, travel motion.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610364871.0A CN105838607A (en) | 2016-05-27 | 2016-05-27 | Cell cultivation insertion component for scratch experiments and application of cell cultivation insertion component |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610364871.0A CN105838607A (en) | 2016-05-27 | 2016-05-27 | Cell cultivation insertion component for scratch experiments and application of cell cultivation insertion component |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105838607A true CN105838607A (en) | 2016-08-10 |
Family
ID=56594829
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610364871.0A Pending CN105838607A (en) | 2016-05-27 | 2016-05-27 | Cell cultivation insertion component for scratch experiments and application of cell cultivation insertion component |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105838607A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520519A (en) * | 2016-11-11 | 2017-03-22 | 四川大学华西医院 | Ruler scratching device used for wound healing cell experiments |
| CN108384715A (en) * | 2018-04-27 | 2018-08-10 | 上海长海医院 | A kind of cell scratch experiment culture dish and its application method |
| 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 |
| CN110408536A (en) * | 2019-06-12 | 2019-11-05 | 上海大学 | Cell migration assay device and method |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120156773A1 (en) * | 2010-12-16 | 2012-06-21 | General Electric Company | Cell carrier, methods of making and use |
| CN204222410U (en) * | 2014-11-17 | 2015-03-25 | 中南大学湘雅二医院 | A kind of seal for doing scratch test to cell |
| CN204509360U (en) * | 2014-12-18 | 2015-07-29 | 华东医院 | A kind of cell scratch experiment incubator |
| CN205635642U (en) * | 2016-05-27 | 2016-10-12 | 上海臻和生物科技有限公司 | A device and consumptive material suit for mar experiment |
-
2016
- 2016-05-27 CN CN201610364871.0A patent/CN105838607A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120156773A1 (en) * | 2010-12-16 | 2012-06-21 | General Electric Company | Cell carrier, methods of making and use |
| CN204222410U (en) * | 2014-11-17 | 2015-03-25 | 中南大学湘雅二医院 | A kind of seal for doing scratch test to cell |
| CN204509360U (en) * | 2014-12-18 | 2015-07-29 | 华东医院 | A kind of cell scratch experiment incubator |
| CN205635642U (en) * | 2016-05-27 | 2016-10-12 | 上海臻和生物科技有限公司 | A device and consumptive material suit for mar experiment |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106520519A (en) * | 2016-11-11 | 2017-03-22 | 四川大学华西医院 | Ruler scratching device used for wound healing cell experiments |
| CN108384715A (en) * | 2018-04-27 | 2018-08-10 | 上海长海医院 | A kind of cell scratch experiment culture dish and its application method |
| CN108384715B (en) * | 2018-04-27 | 2023-04-25 | 上海长海医院 | Culture dish for cell scratch experiment and use method thereof |
| 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 |
| CN110408536A (en) * | 2019-06-12 | 2019-11-05 | 上海大学 | Cell migration assay device and method |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105838607A (en) | Cell cultivation insertion component for scratch experiments and application of cell cultivation insertion component | |
| Anselme et al. | Role of the nucleus as a sensor of cell environment topography | |
| Vedel et al. | Migration of cells in a social context | |
| ATE553187T1 (en) | CONDITIONED CELL CULTURE MEDIUM, METHOD FOR PREPARATION AND USE THEREOF FOR MAINTAINING, PROLIFERATION AND DIFFERENTIATION OF MAMMAL CELLS | |
| CN205635642U (en) | A device and consumptive material suit for mar experiment | |
| CN202658162U (en) | Multi-chamber co-culture device | |
| CN203048919U (en) | Stamp type experimental device for cell migration experiments | |
| CN108384718B (en) | Living single cell in-situ cutting method and device | |
| CN108384715B (en) | Culture dish for cell scratch experiment and use method thereof | |
| CN206486548U (en) | The culture dish of umbilical cord mesenchymal stem cells | |
| Nakao et al. | A method for collecting single cell suspensions using an ultrasonic pump | |
| JP2018143168A (en) | Cell culture carrier | |
| CN104357542A (en) | Method for determining activity of trichophyton rubrum keratinase | |
| JP5868244B2 (en) | Cell culture carrier | |
| CN209854172U (en) | Cell scar healing experimental device | |
| CN2830408Y (en) | Cell culture plate | |
| CN208577725U (en) | A kind of cell scratch experiment culture dish | |
| CN201896176U (en) | Sheet making type cell culture plate | |
| CN207552301U (en) | A kind of removable instrument rack for superclean bench | |
| CN207175974U (en) | A kind of multi-functional cell cultivation equipment | |
| CN204779610U (en) | Cover glass | |
| CN207891327U (en) | A kind of many cells co-cultivation ware frame | |
| CN203593747U (en) | Minisize biological shaking table for cell suspending liquid inoculation | |
| CN103396935A (en) | Biological scaffold material fixing rack and using method thereof | |
| CN219772146U (en) | Cell culture plate |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20160810 |