CN106085848B - A kind of cell co-culture device and preparation method thereof, application method and application - Google Patents
A kind of cell co-culture device and preparation method thereof, application method and application Download PDFInfo
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Abstract
The present invention provides a kind of cell co-culture device and preparation method thereof, application method and applications, the cell co-culture device includes bottom plate, several sheet glass for cell culture are placed on bottom plate, shelter is placed on sheet glass, a part of each sheet glass is blocked by obstructions, it is denoted as region 2, the part that sheet glass is not blocked by obstructions is denoted as region 1, and with the label for 2 range of distinguishable region 1 and region on sheet glass.Cell co-culture device provided by the invention, when in use by by specific cell inoculation at specific position, cell type can be determined according to inoculation position, it does not need to carry out molecular labeling to cell again, the interactional research between different cells can be carried out, bring influences each other the allogenic cell that can be used for studying between different conditions and directly contacting, it may also be used for bring does not influence each other allogenic cell and directly contacting for research.
Description
Technical field
The invention belongs to biological cell culture experiment device fields, and be related to one kind influences between cell and cell for studying
Cell co-culture device and preparation method thereof, application method and application.
Background technique
The organic whole that organism is made of various kinds of cell, by influencing each other between different cells, Coordinated Play is made
With, maintain organ, tissue it is normal, influencing each other between cell has been a hot spot of research.There are phases between different cells
Mutually influence, such as: cell communication between oligodendroglia and neuronal cell influences the formation of myelin, cardiac muscle cell at
Influencing each other between fibrocyte participates in the growth course of heart, the interaction between Dendritic Cells and T lymphocyte
Participate in the antigen deduction process, etc. in immune response.There is also influencing each other between allogenic cell different conditions, such as: view
Apoptotic signal in nethike embrane layer can be spread within the organization, make the normal cell around apoptotic cell that apoptosis occur；Myocardial infarction
In the process, dead signal can be connected by gap and be transmitted, and make the normal cell around dead myocardial cells that apoptosis or bad occur
It is dead, etc..Between cell influencing each other extensively, play a significant role under organism physiology and pathological state, be worth be
The research of system.
At present for the influence between cell, there are many experimental method, mainly co-cultured using cell, study between cell
Influence each other.The experimental method that cell co-cultures can be divided into following a few classes；(1) Transwell co-culture system.Experiment dress
It sets and is divided into upper chamber and lower room, separated, upper interior by one layer of polycarbonate membrane semi-permeable membrane (semi-transparent membrane aperture :~3 μm) therebetween
Cultivate a kind of cell, lower interior culture another kind cell, cell itself cannot by semi-permeable membrane, but cell secretion it is various because
Son can be freely penetrating；This experimental provision is used to study a kind of effect of the cell factor of cell secretion to another cell, no
A kind of cell can be used to study by the cell way of contact, the effect to another cell.(2) two kinds of cells are in a culture plate
Interior growth.Two different kinds of cell is successively inoculated in a culture plate according to a certain percentage, after cultivating a period of time, to two
The marker protein of class cell is dyed by immunohistochemistry, distinguishes the variety classes cell in culture plate, and observation co-cultured
The influence that journey grows cell, this experimental method is for studying a kind of cell by secretion factor or directly contact, to another
The influence of kind cell.(3) two kinds of cells are successively grown in a culture plate.In a kind of cell inoculation to culture plate, culture one
After the section time, the fusion close to 80% is reached between cell, it is then a small amount of again to be inoculated with another cell, two are formed in culture plate
Confluent monolayer cells, lower layer are the first cell, and upper layer is second of cell, this experimental method is for studying between different cells by straight
Contact is to the influence between cell.(4) two kinds of cells are grown in a culture plate.Two different kinds of cell is according to one
Certainty ratio is inoculated in a culture plate, after cultivating a period of time, is collected two kinds of cell differentiations by flow cytometry, with laggard
Row biochemical analysis studies influencing each other between cell.(5) method for using genetic engineering, is co-cultured after marking cell.
After fluorescent protein labeling one of which cell, it is cultivated in a culture plate with other cells, distinguishes two kinds by fluorescence
Cell then observes a series of indexs, studies the influence between two kinds of cells.
It is that current experimental method is not able to satisfy there are also a kind of co-cultivation research although having existed a variety of co-culture methods
's.Such as the project studied at present are as follows: research green fluorescent protein (GFP) is between the cardiac muscle cell to contact with each other
Diffusion process, and current co-culture method is not suitable for this research.Reason is: (1) Transwell co-cultures body
System；The problem of the method, is that the method is suitable for studying the interaction between two kinds of cells, is not present between two kinds of cells
It directly contacts, therefore is not suitable for the above subject.(2) two kinds of cells are inoculated in a culture plate simultaneously by a certain percentage；This
The problem of method, is, if needed in advance by way of virus infection using this co-culture method, GFP is imported cardiac muscle
Into the cell, then the cardiac muscle cell of the cardiac muscle cell of virus infection and uninfecting virus are co-cultured in a culture plate, is seen
Examine the diffusion process of GFP albumen.The defect of the method is that situation there are two types of green fluorescences occurs in late stage of culture cell, the first
Situation: the few cell of virus infection amount, early period, GFP expression quantity was few, cannot be identified in early period, and virus passes through self-replication, after
Phase GFP expression quantity increases, and cell will appear fluorescence；Second situation: early period virus infection, express and GFP and fluorescence occur
Cardiac muscle cell, after being contacted with the cell that surrounding is uninfected by, in several ways, GFP can from the cellular invasion of virus infection to
The cell of the cell of uninfecting virus, uninfecting virus also will appear fluorescence.This co-culture method, which cannot distinguish between, there is fluorescence
Cell is the first situation or second situation, therefore this method is not suitable for the research of the above subject.(3) two kinds of cells
It is co-cultured in a manner of bilevel；The problem of the method, is that cardiac muscle cell, not only can phase after cultivating a period of time
Mutually fusion, can also be grown, the time is longer, and bilevel cell differentiation is more unobvious, therefore this method is not in a manner of stacking
It can be used for the research of the above subject.(4) two kinds of cells are separated and collected by flow cytometry after cell co-cultures；The method is asked
Topic is that the method can only distinguish the cardiac muscle cell of expression GFP and not express the cardiac muscle cell of GFP, not can confirm that GFP is derived from
Virus infection or GFP are from other cellular invasions, therefore this method cannot be used for the research of the above subject.(5) base is used
It is co-cultured after marking cell because of the method for engineering；The problem of the method, is, carries out GFP and second point simultaneously to cell
Son label, it cannot be guaranteed that second of molecule is confined to the cell of virus infection always, it is possible to the side that can be also contacted by cell
Formula transfer, therefore this method cannot be used for the research of the above subject.Therefore need exploitation design is a set of to can be used in the above subject
New cell co-culture device so that do not need to cell carry out molecular labeling, can study of the same race between different conditions
Bring influences each other cell and directly contacting.
Summary of the invention
The purpose of the present invention is to provide a kind of cell co-culture device and preparation method thereof, application method and applications, make
It does not need to carry out molecular labeling to cell when with the cell co-culture device, the allogenic cell that can be studied between different conditions is logical
Cross directly contact and bring influences each other, while can also study not allogenic cell by directly contacting and the mutual shadow of bring
It rings.
In order to achieve the above objectives, the technical solution adopted by the present invention are as follows:
A kind of cell co-culture device, including bottom plate are placed with several sheet glass for cell culture, glass on bottom plate
On piece is placed with shelter, and a part of each sheet glass is blocked by obstructions, and is denoted as region 2, and sheet glass is not blocked object
The part sheltered from is denoted as region 1, and with the label for 2 range of distinguishable region 1 and region on sheet glass.
The bottom plate is the glass plate of surfacing, and sheet glass is coverslip, and shelter is glass slide, coverslip or saturating
Gelatin band.
The production method of the cell co-culture device, the specific steps of which are as follows:
Surfacing, clean bottom plate are chosen, several surfacings, clean sheet glass are placed on bottom plate, and in glass
Glass on piece puts on the label for 2 range of distinguishable region 1 and region, then shelters from region 2 with shelter, then the place that carries out disinfection
It manages to get cell co-culture device is arrived.
The disinfection treatment is sterilized by the way of cobalt source irradiation.
Application of the cell co-culture device in terms of cultivating two different cells.
Application of the cell co-culture device in terms of the allogenic cell under two kinds of culture different cell states.
The application method of the cell co-culture device, the specific steps of which are as follows:
1) sterile cell co-culture device is put into culture dish, then cell A is seeded in culture dish and is trained
Support, then cell co-culture device removed out of culture dish, at this time region uncovered in cell co-culture device insole board,
Cell A has been sticked on region 1 and shelter on sheet glass；
2) shelter is removed from cell co-culture device, exposes the region 2 not sticked by cell A, then remove bottom plate,
Only retain sheet glass, has sticked cell A on the region 1 of sheet glass at this time, do not sticked any cell on region 2；
3) sheet glass is moved into 24 orifice plates, into 24 orifice plates, inoculation carries the virus of gene X, and incubated cell makes virus
Infection cell A, the part cell A in glass panel region 1 is infected at this time, then the culture solution in 24 orifice plates is changed into fresh
Culture solution removes the virus dissociated in culture solution；
4) inoculating cell A is cultivated again in 24 orifice plates, has been sticked on the region 1 and region 2 of sheet glass at this time
The cell A of second of inoculation, with the cell A not being infected and first time for also sticking first time inoculation on time domain 1
The cell A of inoculation being infected；
5) sheet glass is moved into 24 new orifice plates and continues cell culture, the cell sticked on last collecting zone 2
Biochemical analysis is carried out, influence of the cell to 2 inner cell of region in analyzed area 1.
The application method of the cell co-culture device, the specific steps of which are as follows:
1) sterile cell co-culture device is put into culture dish, then cell A is seeded in culture dish and is trained
Support, then cell co-culture device removed out of culture dish, at this time region uncovered in cell co-culture device insole board,
Cell A has been sticked on region 1 and shelter on sheet glass；
2) shelter is removed from cell co-culture device, exposes the region 2 not sticked by cell A, then remove bottom plate,
Only retain sheet glass, has sticked cell A on the region 1 of sheet glass at this time, do not sticked any cell on region 2；
3) sheet glass is moved into 24 orifice plates, into 24 orifice plates, inoculating cell B is cultivated, at this time the region 1 of sheet glass
Above while cell A and cell B are sticked, has only sticked cell B on region 2；
4) sheet glass is moved into 24 new orifice plates and continues cell culture, the cell sticked on last collecting zone 2
Biochemical analysis is carried out, influence of the cell to 2 inner cell of region in analyzed area 1.
Compared with the existing technology, the invention has the benefit that
Cell co-culture device structure provided by the invention is simple, and production method is easy, and is easy to use, and have
Advantage not available for 4 points of existing cell co-culture devices below, specific as follows:
1) cell co-culture device provided by the invention can be used for studying the influence between two kinds of different cells: by cell
A is seeded in region 1, and cell B is seeded in region 2, can study influence of the cell A to cell B, can especially study cell A
The influence that cell B is generated by way of direct exposing cell B；
2) cell co-culture device provided by the invention can also study the influence between allogenic cell different conditions: by A
The cell C of state is seeded in region 1, and the cell C of B state is seeded in region 2, can study the cell C of A condition to B state
The influence of cell C can especially study the cell C of A condition by way of the cell C for directly contacting B state to B state
The influence that cell C is generated；
3) cell co-culture device provided by the invention does not need that cell to be studied is marked before use: because
The application method of cell co-culture device according to the present invention, at specific position by specific cell inoculation, finally in region 2
Interior cell is all same class, after marking the position of determining region 2 on the glass sheet, can determine sheet glass according to position
The type of upper cell, without determining cell type by the marker protein or form of cell itself；
4) after having used cell co-culture device culture cell provided by the invention, the cell in region 2 is received
Collection, can carry out relevant biochemical analysis, and the cell in survey region 1 influences the protein expression of 2 inner cell of region: can incite somebody to action
Cell in region 2 directly carries out fixation in situ, immunohistochemical staining analysis, the metamorphosis and egg of 2 cell of survey region
White changes in distribution in the cell.
The allogenic cell that cell co-culture device provided by the invention can be used in studying between different conditions passes through direct
Contact and bring influences each other, while not allogenic cell can also be studied bring influences each other and directly contacting, can
It is applied in terms of cultivating two different cells, it also can be in terms of the allogenic cell under two kinds of culture different cell states
It is applied, there is good practicability and broad application prospect.
Detailed description of the invention
Fig. 1 is the production process schematic diagram of cell co-culture device provided by the invention；
Fig. 2 is the pictorial diagram for the cell co-culture device that the present invention makes；
Fig. 3 is the use flow diagram of cell co-culture device provided by the invention；
The diffusion process figure of Fig. 4 GFP between myocyte；
Fig. 5 is GFP diffusion process figure between the cardiac muscle cell mutually contacted.
Specific embodiment
The present invention provides a kind of cell co-culture devices, applied to influencing each other between the different cells of research.It is different
Can be influenced each other the function of cell between the cell of type by way of contact, and in such research, part research is used
The mode of fluorescent molecule label distinguishes two kinds of cells, observes a kind of influence of cell to another cell；But another part is ground
In studying carefully, is not suitable for distinguishing two kinds of cells by the way of fluorescent molecule label, certain difficulty is caused to the development of such research.
Cell co-culture device provided by the invention at specific position by specific cell inoculation determines cell according to inoculation position
Type does not need to carry out molecular labeling to cell again, so that it may carry out the interactional correlative study between different cells.
The present invention is described in further details with reference to the accompanying drawing, described is explanation of the invention, rather than is limited.
As shown in Figure 1, cell co-culture device provided by the invention, including bottom plate, it is placed on bottom plate several for thin
The sheet glass of born of the same parents' culture, is placed with shelter on sheet glass, a part of each sheet glass is blocked by obstructions, and is denoted as region
2, the part that sheet glass is not blocked by obstructions is denoted as region 1, and with for 2 model of distinguishable region 1 and region on sheet glass
The label enclosed.Its insole board requires nothing more than surfacing, can select rectangle coverslip 1. or plastic plate, metal plate etc.；
2. sheet glass can select round coverslip；As long as shelter can reach the function of sheltering from segment glass piece, Ke Yixuan
3. with rectangle coverslip or adhesive tape.
As depicted in figs. 1 and 2, the present invention when making cell co-culture device specifically includes the following steps:
1) the rectangle coverslip of clean surface is chosen 1. (as shown in Figure 1A)；
2) 6 round coverslips are uniformly placed 2. rectangle coverslip is 1. upper, 6 round coverslips 2. on stamp mark
Remember (as shown in Figure 1B)；
3) according to round coverslip 2. on label, 2. round coverslip is divided into two regions: region 1 and region 2 are (such as
Shown in Fig. 1 C)；
4) with rectangle coverslip or adhesive tape 3. overlay area 2 (as shown in figure iD)；
5) device is sterilized by cobalt source radiation modality, that is, completes the production of cell co-culture device, the cell made
The pictorial diagram of co-culture device is as shown in Figure 2.
As shown in figure 3, being to study diffusion process of the green fluorescent protein (GFP) between the cardiac muscle cell to contact with each other
Example illustrates the application method of cell co-culture device provided by the invention, the specific steps of which are as follows:
1) sterile cell co-culture device is put into culture dish (as shown in Figure 3A)；
2) neonatal rat myocardial cell is seeded in culture dish, culture for 24 hours, as shown in Figure 3B, at this time in culture dish, rectangle
Coverslip 1. upper uncovered region, round coverslip 2. on region 1 and adhesive tape 3. on sticked Neonatal myocardial
Cell；
3) cell co-culture device is removed out of culture dish, as shown in Figure 3 C, at this time rectangle coverslip 1. on not by
The region of covering, round coverslip 2. on region 1 and adhesive tape 3. on still sticked neonatal rat myocardial cell；
4) 3. adhesive tape is removed from cell co-culture device, as shown in Figure 3D, at this time on cell co-culture device
The region 3. covered by adhesive tape does not stick neonatal rat myocardial cell, has still sticked neonatal rat myocardial cell on other regions；Figure
3E is the corresponding pictorial diagram of Fig. 3 D, by test be able to verify that adhesive tape 3. to round coverslip 2. on region 1 and region 2
Cell stick no influence；
5) 1. from cell co-culture device, only retains 6 round coverslips 2., such as Fig. 3 F
It is shown, at this time round coverslip 2. on region 1 on sticked neonatal rat myocardial cell, it is thin not stick Neonatal myocardial on region 2
Born of the same parents；
6) 2. 6 round coverslips are moved into 6 holes of 24 orifice plates, as shown in Figure 3 G；
7) it is inoculated with the virus for carrying GFP gene into 24 orifice plates, is incubated for for 24 hours, makes virus infection neonatal rat myocardial cell, such as schemes
Shown in 3H, neonatal rat myocardial cell exist only in round coverslip 2. on region 1 on, and part neonatal rat myocardial cell by virus feel
Dye, remainder neonatal rat myocardial cell are not infected；
8) it changes the culture solution in 24 orifice plates into fresh medium, removes the virus dissociated in liquid；
9) it is inoculated with neonatal rat myocardial cell (second of inoculation cardiac muscle cell) in 24 orifice plates, cultivates for 24 hours, as shown in fig. 31,
At this time in 24 orifice plates and round coverslip 2. on region 1 and region 2 on stick the neonatal rat myocardial cell of second of inoculation；
10) 6 round coverslips are moved into 24 new orifice plates, as shown in figure 3j, round coverslip 2. on region 2 on only
The neonatal rat myocardial cell of second of inoculation is sticked, and docile has the suckling mouse not being infected being inoculated with for the first time on region 1
The neonatal rat myocardial cell of cardiac muscle cell, for the first time neonatal rat myocardial cell of inoculation being infected and second of inoculation；And
In the presence of the intersection in region 1 and region 2, the Neonatal myocardial of some inoculation for the first time in region 1 being infected is thin
The case where some second of neonatal rat myocardial cell being inoculated in born of the same parents and region 2 directly contacts with each other；
11) cell continues to cultivate 12d；So far, the cell in the region 1 of round coverslip 2. includes 3 classes: being inoculated with for the first time
Cardiac muscle cell's (virus infection), cardiac muscle cell's (uninfecting virus) of inoculation for the first time, the cardiac muscle cell of second inoculation；Circle
Cell of the shape coverslip 2. in region 2 is the cardiac muscle cell of second of inoculation.
It is observed in real time with fluorescence microscope in the cell state in round coverslip 2. region 2 and the GFP in region 2
Expression process.The cell in region 1 and region 2 is there are cell contact at line of demarcation, and there are cell connection between this two parts cell
It is (as shown in figure 3j).Because cell in region 2 is from not in contact with excessively viral, cell itself will not centainly express GFP, region 1
Interior part cell can express green fluorescence；After cultivating 12d, if having found the cardiac muscle of expression green fluorescence in region 2
Cell, it may be considered that GFP must be passed over from the cell in region 1, it was demonstrated that GFP can pass through certain between cell
Mode is transmitted, and specific experimental result is as shown in Figure 4, Figure 5.
Fig. 4 gives the diffusion process of GFP between cardiac muscle cell, and wherein A-F is cell fluorescence microphoto, and A '-F ' is
Cell differs microphoto.Dotted line Boundary is the following are region 1, and the above are regions 2 by dotted line Boundary.It can be seen that cell
After cultivating 12d, green fluorescence has diffused to the Frontier in region 2.
Fig. 5 gives the diffusion process of GFP between the cardiac muscle cell to contact with each other, and wherein cell 1 is located in region 1, cell 2
In region 2, and cell 1 and cell 2 contact with each other.After cultivating 8d, there is fluorescence in cell 1, and cell 2 does not occur fluorescence；And
After cultivating 12d, there is fluorescence in cell 1 and cell 2.
The advantages of cell co-culture device provided by the invention, is: 1) can study the influence between two kinds of cells；Carefully
Born of the same parents A is seeded in region 1, and cell B is seeded in region 2, studies influence of the cell A to cell B.2) allogenic cell difference can be studied
Influence between state；The cell inoculation of A condition studies the cell of A condition in region 2 in region 1, the cell inoculation of B state
To the impact cell of B state.3) it does not need that cell is marked；Cell in region 2 is all same class, determines that region 2 exists
It is the type that can determine cell according to position, without passing through the marker protein or form of cell itself behind position on coverslip
Determine cell type.4) cell in region 2 can carry out relevant biochemical analysis after collecting, thin in survey region 1
Born of the same parents influence the protein expression of 2 inner cell of region；Cell in region 2 is directly subjected to fixation in situ, immunohistochemical staining point
Analysis, the changes in distribution of the metamorphosis and albumen of 2 cell of survey region in the cell.This 4 points be it is current it has been reported that
Advantage not available for cell co-culture device.But cell co-culture device provided by the invention can not rule out paracrine to thin
The influence of born of the same parents only focuses on the influence that cell by cell is studied by the direct way of contact, therefore using provided by the invention thin
Before born of the same parents' co-culture device, it should first carry out transwell co-culture experiments, exclude influence of the paracrine between cell.
Claims (2)
1. a kind of application method of cell co-culture device, which is characterized in that the cell co-culture device, including bottom plate, bottom
It is placed with several sheet glass for cell culture on plate, shelter, a part of quilt of each sheet glass are placed on sheet glass
Shelter shelters from, and is denoted as region 2, and the part that sheet glass is not blocked by obstructions is denoted as region 1, and with useful on sheet glass
In the label of 2 range of distinguishable region 1 and region；Region 1 and region 2 are respectively used to cultivate different cells；The bottom plate is
The glass plate of surfacing, sheet glass are coverslip, and shelter is glass slide, coverslip or adhesive tape；The cell co-cultures
The application method of device, the specific steps are as follows:
1) sterile cell co-culture device is put into culture dish, then cell A is seeded in culture dish and is cultivated, then
Cell co-culture device is removed out of culture dish, at this time region uncovered in cell co-culture device insole board, glass
Cell A has been sticked on the region 1 of on piece and shelter；
2) shelter is removed from cell co-culture device, exposes the region 2 not sticked by cell A, then remove bottom plate, only protected
Sheet glass is stayed, cell A has been sticked on the region 1 of sheet glass at this time, has not sticked any cell on region 2；
3) sheet glass is moved into 24 orifice plates, into 24 orifice plates, inoculation carries the virus of gene X, and incubated cell makes virus infection
Cell A, the part cell A in glass panel region 1 is infected at this time, then changes the culture solution in 24 orifice plates into fresh cultured
Liquid removes the virus dissociated in culture solution；
4) inoculating cell A is cultivated again in 24 orifice plates, has sticked second on the region 1 and region 2 of sheet glass at this time
The cell A of secondary inoculation is inoculated with the cell A of inoculation not being infected and first time for the first time is also sticked on time domain 1
The cell A being infected；
5) sheet glass is moved into 24 new orifice plates and continues cell culture, the cell sticked on last collecting zone 2 carries out
Biochemical analysis, influence of the cell to 2 inner cell of region in analyzed area 1.
2. a kind of application method of cell co-culture device, which is characterized in that the cell co-culture device, including bottom plate, bottom
It is placed with several sheet glass for cell culture on plate, shelter, a part of quilt of each sheet glass are placed on sheet glass
Shelter shelters from, and is denoted as region 2, and the part that sheet glass is not blocked by obstructions is denoted as region 1, and with useful on sheet glass
In the label of 2 range of distinguishable region 1 and region；Region 1 and region 2 are respectively used to cultivate different cells；The bottom plate is
The glass plate of surfacing, sheet glass are coverslip, and shelter is glass slide, coverslip or adhesive tape；The cell co-cultures
The application method of device, the specific steps are as follows:
1) sterile cell co-culture device is put into culture dish, then cell A is seeded in culture dish and is cultivated, then
Cell co-culture device is removed out of culture dish, at this time region uncovered in cell co-culture device insole board, glass
Cell A has been sticked on the region 1 of on piece and shelter；
2) shelter is removed from cell co-culture device, exposes the region 2 not sticked by cell A, then remove bottom plate, only protected
Sheet glass is stayed, cell A has been sticked on the region 1 of sheet glass at this time, has not sticked any cell on region 2；
3) sheet glass is moved into 24 orifice plates, into 24 orifice plates, inoculating cell B is cultivated, same on the region 1 of sheet glass at this time
When sticked cell A and cell B, only sticked cell B on region 2；
4) sheet glass is moved into 24 new orifice plates and continues cell culture, the cell sticked on last collecting zone 2 carries out
Biochemical analysis, influence of the cell to 2 inner cell of region in analyzed area 1.
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| CN110724637A (en) * | 2019-11-22 | 2020-01-24 | 南通大学 | Glass slide system for direct contact cell co-culture and working method thereof |
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| CN201737950U (en) * | 2010-07-29 | 2011-02-09 | 上海中医药大学 | Special experimental apparatus for cell growing on cover glass |
| CN102273431A (en) * | 2011-06-01 | 2011-12-14 | 深圳大学 | Method for co-culturing freshwater rotifers and chlorella vulgaris |
| CN203999635U (en) * | 2014-07-17 | 2014-12-10 | 天津医科大学总医院 | Detachable cell climbing sheet culturing plate |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN201737950U (en) * | 2010-07-29 | 2011-02-09 | 上海中医药大学 | Special experimental apparatus for cell growing on cover glass |
| CN102273431A (en) * | 2011-06-01 | 2011-12-14 | 深圳大学 | Method for co-culturing freshwater rotifers and chlorella vulgaris |
| CN203999635U (en) * | 2014-07-17 | 2014-12-10 | 天津医科大学总医院 | Detachable cell climbing sheet culturing plate |
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