CN207294812U - A kind of single neuron culture mould for being used to study barometric gradient damage - Google Patents
A kind of single neuron culture mould for being used to study barometric gradient damage Download PDFInfo
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- CN207294812U CN207294812U CN201720070906.XU CN201720070906U CN207294812U CN 207294812 U CN207294812 U CN 207294812U CN 201720070906 U CN201720070906 U CN 201720070906U CN 207294812 U CN207294812 U CN 207294812U
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Abstract
The utility model provides a kind of single neuron culture mould for being used to study barometric gradient damage, the mould includes sheet glass, trapezoidal hydrogel layer and the hydrogel layer containing neuronal cell, the trapezoidal hydrogel layer is shaped to trapezoidal by dovetail groove, and the hydrogel layer containing neuronal cell is located at the head end of trapezoidal hydrogel layer（H）, trapezoidal hydrogel layer and the hydrogel layer containing neuronal cell are freezed solidly on sheet glass.The trapezoidal axon growth hydrogel of " head height tail is low " is prepared by photopolymerization twice in the utility model, and neuronal cell is fixed on to the head end of axon growth gel（H）, in the external force of the exsule long gel direction loading fixed size of vertical axis, gradient stress, the barometric gradient that the long aixs cylinder of imictron is experienced are formed using deformation degree difference after gel stress.
Description
Technical field
The utility model belongs to cellular damage research field, more particularly to a kind of single god for being used to study barometric gradient damage
Mould is cultivated through member.
Background technology
Cell in body experiences a variety of power with location difference（Shearing force, pressure, distraction force
Deng）.After cell stress exceeds tolerance range, damage just occurs, studies this damage process is biomedical sector one
A important topic.In order to study damage of the power to cell, various Mechanical loading systems are developed, wherein the U.S.
Flexcell systems, can complete the loading to cell flow shearing force, pressure, pulling force etc..These loading systems are mostly pins
Mechanical loading is integrally carried out to cell, i.e., the power that individual cells are subject in the same time is identical, can be met most
Research.But the Shortcomings in neuron stress research.Partial nerve member president in nervous system goes out the aixs cylinder of overlength（Centimetre
Level）, the stress of such individual cells just becomes line from point, i.e. individual cells different parts are subject to various forces.It is the most typical
Example be exactly retinal ganglial cells（RGC）, its cell space part is located in eyeball, is subject to intraocular pressure；And its aixs cylinder is located at
With the sealed tube intracavitary of ventricles of the brain unicom, it is subject to the pressure of cerebrospinal fluid, and intraocular pressure is different from intracranial pressure, can form a pressure
Gradient, when intraocular pressure and the cranium pressure great change of generation, it may occur that irreversible optic nerve injury, causes to blind.
The content of the invention
To solve the above problems, the utility model is intended to establish a single neuron training for being used to study barometric gradient damage
A kind of system of supporting, there is provided single neuron culture mould for being used to study barometric gradient damage.
The purpose of this utility model is achieved through the following technical solutions：
It is a kind of be used for study barometric gradient damage single neuron culture mould, including sheet glass, trapezoidal hydrogel layer and
Hydrogel layer containing neuronal cell, the trapezoidal hydrogel layer be shaped to by dovetail groove it is trapezoidal, it is described to contain neuronal cell
Hydrogel layer be located at the trapezoidal head end H of trapezoidal hydrogel layer, trapezoidal hydrogel layer and the hydrogel layer containing neuronal cell coagulate
It is fixed on sheet glass.
Further, the sheet glass is via the processed lid glass of 3- (trimethoxy first silicon substrate) propyl methacrylates
Piece.
Further, the hydrogel layer（3）With the hydrogel layer containing neuronal cell（5）Prepared by hydrogel solution
Obtain, the hydrogel solution is the GelMA hydrogels of concentration 3% ~ 8%, and wherein contains 0.1% ~ 1% photosensitizer.
Further, the GelMA hydrogels are prepared by phosphate buffer solution.
The preparation method of the single neuron culture mould for being used to study barometric gradient damage, comprises the following steps：
1）The preparation of photomask
Prepare axon growth photomask（1-1）With cell capture point photomask（1-2）；
2）The preparation of hydrogel solution
GelMA is dissolved into phosphate buffer solution, configures the GelMA hydrogel solutions of 3% ~ 8% final concentration, and is added
0.1% ~ 1% photosensitizer, obtains hydrogel solution；
3）The preparation of trapezoidal GelMA hydrogels
By step 2）The hydrogel solution being prepared is circulated into the dovetail groove being coated with by poly-D-lysine, above according to
Secondary cover glass piece, axon growth photomask, then photopolymerization is carried out with ultraviolet light, obtain the trapezoidal hydrogel layer of axon growth；
4）The preparation of GelMA hydrogel single cell suspensions
Extracorporeal neuron is resuspended to concentration in 3% ~ 8% GelMA hydrogel solutions, to obtain the GelMA containing neuron and hanging
Liquid；
5）The unicellular capture of neuron
By step 4）The prepared GelMA suspensions containing neuron are filled into rectangle poly-D-lysine substrate groove, will be walked
Rapid 3）The hydrogel layer of obtained axon growth is placed in rectangular channel, trapezoidal hydrogel layer is dipped in the GelMA containing neuron and is hanged
In liquid, axon growth photomask is changed to cell capture point photomask, then photopolymerization is carried out with ultraviolet light, completes neuron list
Cell capture, that is, obtain the single neuron culture mould for being used to study barometric gradient damage.
Further, the photomask is film material, the axon growth photomask（1-1）Printing opacity at it is rectangular,
The cell capture point photomask（1-2）Printing opacity at it is rounded.
Further, axon growth photomask（1-1）Rectangle printing opacity at length and width be respectively 1cm, 20 μm, cell capture
Point photomask（1-2）Circular printing opacity at a diameter of 150 μm.
The utility model having the beneficial effect that compared with prior art：
1st, the utility model uses gelatin-methacrylate（GelMA）Material, by adding photosensitizer, makes it have
Photopolymerization ability, by first time photopolymerization, is prepared the trapezoidal axon growth hydrogel of " head height tail is low ", passes through second
Neuronal cell, is fixed on the head end of axon growth gel by photopolymerization（H）, long aixs cylinder neuron is obtained after in vitro culture
Cell, in the external force of the exsule long gel direction loading fixed size of vertical axis, utilizes deformation degree difference institute shape after gel stress
Into gradient stress, barometric gradient that the long aixs cylinder of imictron is experienced.
Brief description of the drawings
Fig. 1 is axon growth photomask（1-1）Structure diagram；
Fig. 2 is cell capture point photomask（1-2）Structure diagram；
Fig. 3 prepares schematic diagram for trapezoidal hydrogel；
Fig. 4 is unicellular capture schematic diagram；
Fig. 5 is the single neuron culture mold structure diagram for studying barometric gradient damage being prepared.
Embodiment
Present embodiments provide a kind of single neuron culture mould for being used to study barometric gradient damage, its preparation method
For：
1）The preparation of photomask
Prepare axon growth photomask 1-1 and cell capture point photomask 1-2, the photomask as shown in Figure 1 and Figure 2, axis
It is rectangular at the printing opacity of exsule long photomask 1-1, length and width be respectively 1cm, 20 μm；At the printing opacity of cell capture point photomask 1-2
It is rounded, a diameter of 150 μm；The photomask is film material；
2）The preparation of hydrogel solution
GelMA is dissolved into phosphate buffer solution（DPBS）In, the GelMA hydrogel solutions of 5% final concentration of configuration, and
0.5% photosensitizer is added, obtains hydrogel solution；
The photosensitizer is Irgacure 2959；
3）The preparation of trapezoidal GelMA hydrogels
As shown in figure 3, by step 2）The hydrogel solution being prepared is circulated into the ladder being coated with by poly-D-lysine 4-1
In shape groove, cover glass piece 2, axon growth photomask 1-1 successively, then use 2.95mW/cm above2The ultraviolet light of energy carries out light
Polymerization, obtains the trapezoidal hydrogel layer 3 of axon growth；
The sheet glass is 3- (trimethoxy first silicon substrate) propyl methacrylate（TMSPMA）Processed coverslip；
4）The preparation of GelMA hydrogel single cell suspensions
By extracorporeal neuron, according to 0.7x106/ ml concentration is resuspended to concentration in 5% GelMA hydrogel solutions, to obtain
GelMA suspensions containing neuron；
5）The unicellular capture of neuron
As shown in figure 4, by step 4）The prepared GelMA suspensions containing neuron are filled into rectangle poly-D-lysine substrate
In 4-2 grooves, by step 3）The hydrogel layer of obtained axon growth is placed in rectangular channel, trapezoidal hydrogel layer is dipped in containing nerve
In the GelMA suspensions of member, axon growth photomask is changed to cell capture point photomask, then use 2.95mW/cm2The purple of energy
Outer light carries out photopolymerization, completes the unicellular capture of neuron, the hydrogel layer 5 containing neuronal cell is formed, as shown in figure 5, most
The single neuron culture mould for being used to study barometric gradient damage is obtained eventually.
Claims (1)
1. a kind of single neuron culture mould for being used to study barometric gradient damage, it is characterised in that the mould includes glass
Piece（2）, trapezoidal hydrogel layer（3）With the hydrogel layer containing neuronal cell（5）, the trapezoidal hydrogel layer determined by dovetail groove
Shape is trapezoidal, the hydrogel layer containing neuronal cell（5）Positioned at trapezoidal hydrogel layer（3）Head end（H）, trapezoidal hydrogel
Layer and the hydrogel layer containing neuronal cell freeze solidly on sheet glass（2）On.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720070906.XU CN207294812U (en) | 2017-01-20 | 2017-01-20 | A kind of single neuron culture mould for being used to study barometric gradient damage |
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201720070906.XU CN207294812U (en) | 2017-01-20 | 2017-01-20 | A kind of single neuron culture mould for being used to study barometric gradient damage |
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| CN207294812U true CN207294812U (en) | 2018-05-01 |
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