CN109234143A - A kind of cell scoring devices and its application method - Google Patents
A kind of cell scoring devices and its application method Download PDFInfo
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- CN109234143A CN109234143A CN201811288336.7A CN201811288336A CN109234143A CN 109234143 A CN109234143 A CN 109234143A CN 201811288336 A CN201811288336 A CN 201811288336A CN 109234143 A CN109234143 A CN 109234143A
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Abstract
The invention discloses a kind of cell scoring devices and its application methods, the cell scoring devices are made of several piece barrier piece arranged in parallel, the distance between adjacent barrier piece is equal, by being fixedly connected at the top of barrier piece with the vertically disposed several connection straps of barrier piece between barrier piece, position corresponding with connection strap is equipped with groove at the top of barrier piece, the connection strap is placed in groove, is equipped with several through-holes in the lower section of each barrier piece.Cell scoring devices disclosed by the invention design it is simple, low in cost, easy to use, without additional device auxiliary or increase experiment flow and reusable, when carrying out scratch experiment, the svelteness of acquisition is beautiful, experimental error is small, meet requirement of experiment, provides experiment basis for more accurate detection tumor cell invasion ability.
Description
Technical field
The invention belongs to field of cell culture, more particularly to a kind of cell scoring devices and its application method.
Background technique
Cell culture refers to simulated in vivo environment (sterile, preference temperature, pH value and certain nutritional condition etc.) in vitro,
It is allowed to survive, grow, breed and maintain a kind of method of primary structure and function.Cell culture technology day crescent at this stage
It is different, wherein cell scratch experiment is a kind of method of simple and easy detection cell movement, and experimental cost is low, can be used to examine
Survey the invasion transfer ability of adherent growth tumour cell.The method that cell scratch experiment uses manual drawing straight line more at present, i.e.,
By cell seeding in culture dish, anti-tumor drug is added after cell covers with, then draws in culture dish and takes several straight lines, is gone
Except the cell fragment in scratch, the tumour cell after observing drug effect puts the invasion feelings in scored area in different times
Condition, and invasion area of the tumour cell in scratch is calculated, to evaluate the tumor cell invasion energy after drug effect
Power.
But the conventional method is increasingly difficult to meet the requirement of cell scratch experiment, disadvantage shows following side
Face: (1) lines are bent, and the adherent tighter tumour cell of residual is easy in scratch；(2) manual scratch is easy to lead there are blindness
Cause accuracy that is irregular and influencing experimental data of crossing；(3) manual operations is cumbersome, there is a possibility that increasing cell contamination；(4)
If experiment accuracy is under suspicion, a large amount of scientific research human and material resources and financial resources are wasted.In recent years, the scientific research of many heavyweights
Magazine is because the method for manual drawing straight line is easy to influence the accuracy of experimental data, and refuses to receive the section in relation to such experiment
Grind paper.The scientific research personnel for being engaged in tumor research has a profound understanding of cell scratch experiment significance in tumor invasion.
Therefore, it needs to design one kind scoring devices that are novel, practical and meeting requirement of experiment to solve existing skill
The problem of art, this also will be helpful to the development of tumor invasion Journal of Sex Research.
Summary of the invention
In view of the above problems, the present invention is intended to provide a kind of cell scoring devices and its application method, structure letter
It is single, it is easy to use, it is the svelteness of acquisition, beauty, neat, experiment is provided for more accurate detection tumor cell invasion ability
Basis.
To achieve the goals above, the technical solution adopted in the present invention is as follows: a kind of cell scoring devices, the cell
Scoring devices are made of several piece barrier piece arranged in parallel, and the height of barrier piece is lower than the height of culture dish, adjacent barrier piece it
Between be equidistant, by being fixedly connected at the top of barrier piece with the vertically disposed several connection straps of barrier piece between barrier piece, every
Position corresponding with connection strap is equipped with groove at the top of baffle, and the connection strap is placed in groove, sets below each barrier piece
There are several through-holes, the specific location of through-hole can not also do specific limit at middle part, the shape of through-hole in the end of barrier piece yet
It is fixed, it is round, rectangular, as long as using when culture solution can be made i.e. applicable by through-hole.
Further, the production material of the barrier piece and connection strap includes glass, stainless steel or plastics, herein to accessory
Material is not particularly limited, as long as can meet the isolation fixed function of barrier piece and connection strap respectively.
Further, the barrier piece includes glass glue connection with the mode that is fixedly connected between connection strap, and glass cement is fixed
Only one of connection type, other connection types can also be with, as long as being able to achieve being fixedly connected for barrier piece and connection strap
?.
Further, when the barrier piece quantity is odd number, the length of intermediate barrier piece and the inner ring diameter of culture dish
It is adapted, the symmetrical setting of barrier piece of two sides, and the length of barrier piece is sequentially reduced from centre to two sides, every piece of side barrier
The length profile of culture dish inner ring is adapted in the length of piece and the barrier leaf length extending direction.
Further, when the barrier piece quantity is even number, the symmetrical setting of barrier piece, the length of barrier piece from centre to
Two sides are sequentially reduced, and the length of every piece of barrier piece and the length profile of culture dish inner ring on the barrier leaf length extending direction are mutually fitted
It answers.
A kind of application method of cell scoring devices, specifically used step are as follows:
1) before cell inoculation, cell scoring devices are first subjected to autoclave sterilization in autoclave, then deposited in super
It is spare in net platform；
2) cell scoring devices are placed in culture dish, confirmation device is put neatly, and pairing is tight, then by processed logarithm
The cell inoculation in growth period is cultivated in incubator after being mixed the cell piping and druming in culture dish with suction pipe in culture dish；
3) when it is 50 ~ 60% that cell, which grows to density, drug-treated is carried out, selectivity replaces culture medium, continues to be placed in incubator
Middle culture；
4) when cell density is 80 ~ 90%, cell scoring devices is removed, anti-tumor drug is then added, removing device
Different time sections afterwards, through related drugs treated invasive ability, and are photographed to record in microscopically observation tumour cell, then
Different time points cell invasion area is compared according to requirement of experiment, obtains experiment conclusion.
The beneficial effects of the present invention are:
It is the svelteness of acquisition, beauty, neat and straight 1. carry out scratch experiment using cell scoring devices disclosed by the invention
Diameter is equal, under the microscope observing effect are as follows: the diameter between straight, rule, different lines is identical, and lines hecatomeral cells is equal
It is even；Because being equidistant between each straight line, drug concentration used, cell density are identical, and experimental error is small, meets requirement of experiment；
Cell gradually covers with scored area after removing device, provides experiment base for more accurate detection tumor cell invasion ability
Plinth；
2. the design of cell scoring devices is simple, the barrier piece group of several pieces of different lengths is selected according to the difference of culture dish internal diameter
At cheap；It is simple, convenient when use, without additional device auxiliary or increase experiment flow；
3. cell scoring devices Preservation in sterile condition after conventional autoclave sterilization strictly observes cell culture process, carefully in super-clean bench
The probability of born of the same parents' pollution is small and reusable；
4. cell scoring devices are easy to use: before inoculating cell enters culture dish, the cell scoring devices after autoclave sterilization being put
Enter in culture dish, inoculates cell and enter culture dish；When cell grows to the density for meeting requirement of experiment, device is removed
Carry out microscopically observation.
Detailed description of the invention
Fig. 1 is the structural schematic diagram of barrier piece disclosed in the embodiment of the present invention 1；
Fig. 2 is the structural schematic diagram of cell scoring devices disclosed in the embodiment of the present invention 1；
Fig. 3 is the schematic diagram of experimental group scratch after removing cell scoring devices of the embodiment of the present invention 3；
Fig. 4 be the embodiment of the present invention 3 experimental group after removing cell scoring devices 0 hour cell growth figure；
Fig. 5 be the embodiment of the present invention 3 experimental group after removing cell scoring devices 24 hours cell growth figures；
Fig. 6 be the embodiment of the present invention 3 experimental group after removing cell scoring devices 48 hours cell growth figures；
Fig. 7 be the embodiment of the present invention 3 control group scratch after 48 hours cell growth figures.
Wherein, 1- barrier piece, 2- connection strap, 3- through-hole, 4- cell, 5- scratch straight line.
Specific embodiment
In order to make those skilled in the art be better understood on technical solution of the present invention, with reference to the accompanying drawing and
Embodiment is further described technical solution of the present invention.
Embodiment 1
Cell scoring devices disclosed in the present embodiment are used for 6 hole culture dishes, and specific structure is as shown in Figs. 1-2, including 5 pieces of barriers
Piece, the height of barrier piece are lower than the height of culture dish, are parallel to each other between barrier piece, the spacing between adjacent barrier piece is equal, position
In intermediate barrier piece longest, other 4 pieces of barrier pieces are symmetrical arranged at left and right sides of it, length is successively passed from centre to two sides
Subtract, be fixedly connected between barrier piece by several connection straps of setting perpendicular at the top of barrier piece, at the top of barrier piece with connection
The corresponding position of item is equipped with groove, and the connection strap is placed in groove, and the lower position of every piece of barrier piece front and back end is equipped with through-hole；
Design parameter are as follows: intermediate 34.5 mm of barrier length of a film, the barrier length of a film 27.6mm in its two sides adjacent position, outermost barrier piece
Long 12.2mm.
Embodiment 2
Cell scoring devices disclosed in the present embodiment include 6 pieces of barrier pieces and 3 connection straps, the bottom center position of every piece of barrier piece
It sets and is all provided with through-hole, circulate for culture solution, barrier piece is parallel to each other, and the height of barrier piece is lower than the height of culture dish, adjacent
The spacing of barrier piece is equal, and barrier piece is symmetrically arranged in two sides, length is successively successively decreased arrangement from centre to two sides, logical between barrier piece
Three connection straps for crossing perpendicular setting at the top of barrier piece are fixedly connected, and position corresponding with connection strap is set at the top of barrier piece
Fluted, the connection strap is placed in groove, and barrier piece and connection strap are fixed by glass cement.
Embodiment 3
For cultivating lung cell A549 and be inoculated in 6 orifice plates progress cell scratch experiment:
Experimental group:
1, before A549 cell inoculation, cell scoring devices disclosed in embodiment 1 are first subjected to high pressure in autoclave
Disinfection, is then deposited in spare in super-clean bench；
2, the A549 cell in logarithmic growth phase is discarded into culture medium, with 1 × PBS(phosphate buffered saline solution) it rinses 2 times,
0.25% pancreatin digests 3 ~ 5 minutes, cell is digested from culture dish bottom, suction pipe is sucked in 15 mL centrifuge tubes, with room
Temperature, 500 rpm centrifugation, 5 minutes collection cells carry out cell count；
3, cell scoring devices are placed in 6 orifice plates, confirmation device is put neatly, and pairing is tight, then by 1 ~ 2 × 105It is a
A549 cell inoculation (adjusts) in 6 orifice plates according to different requirement of experiment, blows and beats the cell in culture dish with suction pipe and mixes
It is cultivated in the incubator of 37 DEG C of+5%CO2 afterwards；
4, when cell grow to density be 50 ~ 60% when, carry out drug-treated, replace culture medium or need not replacement culture medium (according to
Different experiments require adjustment), continue to be placed in incubator and cultivate；
5, when cell density is 80 ~ 90%, cell scoring devices are removed, it is seen that culture dish bottom is clear, neat, beautiful, straight
The equal scratch lines of diameter, as shown in figure 3,4 be the cell in culture dish, 5 be after cell scoring devices remove is in culture dish
Existing scratch straight line；Then anti-tumor drug is added, 0 hour after removing device, 6 hours, 12 hours, 24 hours, 36 hours,
48 hours (being adjusted according to different requirement of experiment) in microscopically observation tumour cell through related drugs treated invasion energy
Power, and photograph to record, different time points cell invasion area is compared further according to requirement of experiment, obtains experiment conclusion.
Control group do not use cell scoring devices, using traditional manual drawing straight-line method scratch, other steps with it is above-mentioned
Method is identical.
Experimental group removes 0 hour after device cell growth status such as Fig. 4, it is seen that and scratch is neat, clear, width is uniform,
Scratch two sides have no obvious cellular damage, and obvious cell intrusion is had no in scratch；Experimental group removes 24 hours after device cells
Growing state such as Fig. 5, it is seen that tumour cell starts to invade or shift into scratch after dosing；Experimental group removes after device 48 hours
Cell growth status such as Fig. 6, it is seen that scratch inner tumour cell has covered with；48 hours cells grow feelings after control group scratch
Condition such as Fig. 7, it is seen that scratch inner cell has no obvious growth.
Therefore, using cell scoring devices of the invention, when the cell in culture dish grows to corresponding density, dress is removed
It sets, visible clear, straight lines in microscopically observation plate, and lines hecatomeral cells is uniform, cell, which gradually covers with, later draws
Trace region provides experiment basis for more accurate detection tumor cell invasion ability.
Basic principles and main features and advantage of the invention have been shown and described above.But the foregoing is merely this hairs
Bright specific embodiment, technical characteristic of the invention are not limited thereto, and any those skilled in the art is not departing from this hair
The other embodiments obtained under bright technical solution should all cover within the scope of the patent of the present invention.
Claims (6)
1. a kind of cell scoring devices, which is characterized in that the cell scoring devices are made of several piece barrier piece arranged in parallel,
The height of barrier piece is lower than the height of culture dish, and the distance between adjacent barrier piece is equal, by the top of barrier piece between barrier piece
It being fixedly connected with the vertically disposed several connection straps of barrier piece, position corresponding with connection strap is equipped with groove at the top of barrier piece,
The connection strap is placed in groove, is equipped with several through-holes in the lower section of each barrier piece.
2. a kind of cell scoring devices as described in claim 1, which is characterized in that the production material of the barrier piece and connection strap
Matter includes glass, stainless steel and plastics.
3. a kind of cell scoring devices as claimed in claim 2, which is characterized in that the fixation between the barrier piece and connection strap
Connection type includes glass glue connection.
4. a kind of cell scoring devices as claimed in claim 3, which is characterized in that the barrier piece quantity is odd number, intermediate
The length of barrier piece be adapted with the inner ring diameter of culture dish, the symmetrical setting of barrier piece of two sides, and the length of barrier piece
It is sequentially reduced from centre to two sides, culture dish inner ring in the length of the every piece of barrier piece in side and the barrier leaf length extending direction
Length profile is adapted.
5. a kind of cell scoring devices as claimed in claim 3, which is characterized in that the barrier piece quantity is even number, barrier
The symmetrical setting of piece, the length of barrier piece are sequentially reduced from centre to two sides, the length of every piece of barrier piece and the barrier leaf length
The length profile of culture dish inner ring is adapted on extending direction.
6. a kind of application method of cell scoring devices according to any one of claims 1 to 5, which is characterized in that specifically make
With step are as follows:
1) before cell inoculation, cell scoring devices are first subjected to autoclave sterilization in autoclave, then deposited in super
It is spare in net platform；
2) cell scoring devices are placed in culture dish, confirmation device is put neatly, and pairing is tight, then by processed logarithm
The cell inoculation in growth period is cultivated in incubator after being mixed the cell piping and druming in culture dish with suction pipe in culture dish；
3) when it is 50 ~ 60% that cell, which grows to density, drug-treated is carried out, selectivity replaces culture medium, continues to be placed in incubator
Middle culture；
4) when cell density is 80 ~ 90%, cell scoring devices is removed, anti-tumor drug is then added, removing device
Different time sections afterwards, through related drugs treated invasive ability, and are photographed to record in microscopically observation tumour cell, then
Different time points cell invasion area is compared according to requirement of experiment, obtains experiment conclusion.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811288336.7A CN109234143A (en) | 2018-10-31 | 2018-10-31 | A kind of cell scoring devices and its application method |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201811288336.7A CN109234143A (en) | 2018-10-31 | 2018-10-31 | A kind of cell scoring devices and its application method |
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| Publication Number | Publication Date |
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| CN109234143A true CN109234143A (en) | 2019-01-18 |
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| CN201811288336.7A Pending CN109234143A (en) | 2018-10-31 | 2018-10-31 | A kind of cell scoring devices and its application method |
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Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040214313A1 (en) * | 2003-04-28 | 2004-10-28 | Weihua Zhang | Cell interaction culture system and uses thereof |
| CN206562441U (en) * | 2017-03-14 | 2017-10-17 | 新疆维吾尔自治区人民医院 | A kind of occupy-place formula cell scratch test device |
| 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 |
| CN209098696U (en) * | 2018-10-31 | 2019-07-12 | 扬州大学 | A kind of cell scoring devices |
-
2018
- 2018-10-31 CN CN201811288336.7A patent/CN109234143A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20040214313A1 (en) * | 2003-04-28 | 2004-10-28 | Weihua Zhang | Cell interaction culture system and uses thereof |
| CN206562441U (en) * | 2017-03-14 | 2017-10-17 | 新疆维吾尔自治区人民医院 | A kind of occupy-place formula cell scratch test device |
| 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 |
| CN209098696U (en) * | 2018-10-31 | 2019-07-12 | 扬州大学 | A kind of cell scoring devices |
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