CN106399078B - A kind of foreign cell 3 D-printing system and method - Google Patents
A kind of foreign cell 3 D-printing system and method Download PDFInfo
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- CN106399078B CN106399078B CN201510455765.9A CN201510455765A CN106399078B CN 106399078 B CN106399078 B CN 106399078B CN 201510455765 A CN201510455765 A CN 201510455765A CN 106399078 B CN106399078 B CN 106399078B
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- 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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
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- 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
Abstract
The present invention relates to a kind of foreign cell 3 D-printing systems, including motion platform, the extruded type cell printing mechanism being fixed on the motion platform and drip-type cell printing mechanism, for observing and feeding back the observing unit and control unit of cell printing situation；Described control unit controls the printing that extruded type born of the same parents' printing mechanism completes three-dimensional cell hydrogel structure first, then controls the printing that drip-type cell printing mechanism completes single or multiple cells on the three-dimensional cell hydrogel structure；The present invention is integrated with micro- extruded type printing technique and the drip-type cell drip-type cell printing technology based on the stagnant inertia force driving of alternation, both the efficient building of macroscopical cell cluster had been ensure that, the guaranteed building of microcosmic individual cells microenvironments near cluster cell again, a kind of external foreign cell model building method for taking into account efficiency and precision is created, there are important pathology, pharmacology application potential.
Description
Technical field
The invention belongs to biology manufacture and cell printing fields, particularly relate to a kind of precisely controlled assembling of foreign cell
Equipment and technique.
Background technique
In recent years, cell printing technology is in organizational engineering, pathological model building, drug screening and detection, cell behavior
The fields such as research play increasingly important role.In field of tissue engineering technology, cell printing technology realizes heterogenous cell
Three-dimensional controlled assembling can construct the structural body of complex organization in vitro, promote organizational project to scale, automation,
The direction of Computer aided building is developed；Field is constructed in pathological model, cell printing can (such as tumour be thin by sick cell
Born of the same parents) assembling of particular order is carried out with normal cell, cell carcinogenesis mechanism, which is studied, for medical worker provides effective research
Means；In drug screening and context of detection, using the cell chip that cell printing technology manufactures can reduce drug screening at
Originally, shorten the period of drug screening and reduce zoopery etc.；In terms of cell behavior research, also using cell printing technology
The cell of particular types and quantity can be assembled and be arranged, thus for rows such as proliferation, migration and the differentiation of research cell
To provide approach.
External three-dimensional cell model it is artificial constructed be international bio medicine and engineering discipline research newest forward position.2000
Year has been inquired into for the first time in " RP (rapid prototyping) is in biomedical Application meeting " that German Freiburg university holds with RP
Based on tissue engineering bracket and Organ printing technology (cell/organ printing) development.Then with histoorgan
The new work repaired and for the purpose of reconstruction, develop the various cell three-dimensionals based on droplet ejection directly controlled assembling and Organ printing
Skill.It is squeezed including the successive cell based on micro-nozzle for tissue construction；Low temperature depositing tissue engineering bracket manufacturing process；Base
In the cell printing of ink-jet technology；The laser direct writing system of cell and bacterium；The micro-contact printing of cell and bacterium；Pass through machinery
, the cell operation that optical, electricity, magnetic, ultrasonic and ion method carries out and by photograph or electroetching with
And the cell that soft lithography carries out is imitated.However tissue system is extremely complex, such as: in-vivo tumour tissue generally has different
Matter, so that the researchs such as oncology, pharmacology, drug development and clinical treatment is made to become unprecedented complexity.Current cell body
The artificial constructed technology of outer three-dimensional cell model is all made of single cell printing mode, and single cell printing mode exists
Respective drawback if the successive cell squash type of micro-nozzle is conducive to the whole cell framework of building, and is unable to complete drip-type
The fixed point of cell printing is sprayed, and the mode of the laser direct writing system of cell and bacterium is conducive to complete the single printing of cell, but
In the building for the overall architecture for being unable to complete cell.It, just can be by different printing types if different printing types is combined
The advantages of combine, the printing of the bionical complex three-dimensional heterogeneous model of height is completed, to meet to many cells labyrinth
The bionical building requirement of tissue.But the driving for being different printing type is different, and control unit is also completely different, therefore at present also
The cell printing equipment that printing type of two kinds of different drivings is not combined.
Summary of the invention
(1) technical problems to be solved
For in the prior art can not be by the combined printing device of the printing type of two kinds of different drivings, and nothing
Method completes the defect of foreign cell 3 D-printing, and the present invention provides a kind of printing type by two kinds of different drivings and is combined together
Foreign cell 3 D-printing system.
(2) technical solution
Foreign cell 3 D-printing system of the present invention including motion platform, is fixed on the motion platform
Extruded type cell printing mechanism and drip-type cell printing mechanism, for observe and feed back the observing unit of cell printing situation with
And control unit；
Described control unit controls the printing that extruded type cell printing mechanism completes three-dimensional cell hydrogel structure first, so
Control drip-type cell printing mechanism completes the printing of single or multiple cells on the three-dimensional cell hydrogel structure afterwards；?
During printing, described control unit controls the three-dimensional motion of the motion platform, to realize the printing in required position.?
During printing, motion platform need to be in the movement of XY axis and Z-direction progress higher motion precision, extruded type cell printing
It needs to carry out the output of continuous signal, the output of pulse signal in carrying out, control of the present invention is needed in drip-type cell printing
Unit processed can meet above-mentioned three kinds of targets, and stability with higher and precision.
In the present invention, described control unit includes industrial personal computer, PMAC controller, signal generator and power amplifier；
The industrial personal computer is connected with the PMAC controller and the observing unit, for receive feedback information and to
PMAC controller issues instruction；The three-dimensional motion of the PMAC controller control motion platform and the extruded type cell printing machine
Extrusion；Further, the PMAC controller also adjusts the position of drip-type cell printing mechanism, by controlling signal
Device and power amplifier print to control the drip-type of cell.
In the present invention, extruded type cell printing mechanism includes the syringe being fixed on the Z axis of the motion platform
Driving motor, pressure head, syringe and its spray head and heat preservation sleeve, wherein the spray head is mounted on syringe oral area, and syringe is solid
It is scheduled in heat preservation sleeve；Injector drive motor pushes syringe under the control of the PMAC controller, through the pressure head
The continuous extrusion of piston realization internal liquid.
In the present invention, drip-type cell printing mechanism includes that the position be fixed on the Z axis of the motion platform is micro-
Adjust motor, pressure head, reciprocating mechanism and the drip-type cell printing mini sprinkler being attached thereto；Wherein, the mini sprinkler is solid
It is scheduled on reciprocating mechanism, the reciprocating mechanism is successively connected with pressure head and fine position motor；The PMAC control
Device is connected with fine position motor, for finely tuning the position of the mini sprinkler；The PMAC controller also control signal generator
Waveform is generated, the waveform is transported on the reciprocating mechanism after power amplifier amplification, passes through reciprocating motion
Drip-type printing is realized in the movement of mechanism.
Preferably, reciprocating mechanism is selected as encapsulation piezoelectric ceramics.It is generated after receiving electric signal using piezoelectric ceramics
It the characteristics of elongation and contraction, drives mini sprinkler to realize and moves back and forth, realize liquid using the friction and inertia power in reciprocating motion
The drippage of drop.
In the present invention, the observing unit includes horizontal microscope for observing and vertical microscope for observing.Wherein, horizontal micro-
Mirror is observed spray head working depth and the arrangement of spray head inner cell, and vertical microscope positions the cell in hydrogel.
Pass through two microscopes simultaneously carried out from both direction from, on the one hand can to avoid the visual field error caused by oblique view,
Improve the precision of observation；On the other hand the droplet position that the result obtained according to observation can print drip-type is determined,
Provide a kind of profile-followed Method of printing of Real-time Feedback printed according to cell distribution in hydrogel.
It is a further object of the present invention to provide carry out foreign cell 3 D-printing using print system of the present invention
Method, which comprises the steps of:
1) sterilization treatment is carried out to the foreign cell 3 D-printing system；
2) according to the model being pre-designed, described control unit controls Z axis height and the position XY of the motion platform, and
It controls extruded type cell printing mechanism to be printed, obtains three-dimensional cell hydrogel structure；
3) according to the information for model and the observing unit feedback being pre-designed or according only to the model being pre-designed, the control
Unit processed adjusts Z axis height and the position XY of the motion platform, and by the fine tuning to drip-type cell printing mechanism position,
Realize that the position of double spray head working condition switchings and drip-type cell printing positions；
4) described control unit controls drip-type cell printing mechanism and passes through the cell based on alternation stagnant inertia force driving
Printing technique carries out drip-type printing on the three-dimensional cell hydrogel structure, obtains foreign cell threedimensional model；
5) it is needed for different model, step 3) and 4) is repeated, until constructing required foreign cell threedimensional model.
In the present invention, the condensation of hydrogel is using physical crosslinking and chemical crosslinking, institute in micro- extruded type cell printing
It states in physical crosslinking, crosslinks the components of gelatin in the three-dimensional cell hydrogel structure by low temperature, become gel-like
State；In the chemical crosslinking, it is crosslinked the seaweed acid constituents in the three-dimensional cell hydrogel structure by calcium ion；Tool
Body, during the chemical crosslinking, 2~4% CaCl is added into the three-dimensional cell hydrogel structure that printing obtains2
Solution extremely submerges, and keeps 2~4min in room conditions, then siphons away solution, just realizes the three-dimensional cell hydrogel
The chemical crosslinking of structure.The diameter of water-setting collodion silk is between 100-800 microns after printing.
In the present invention, the drip-type cell printing is immediately begun to after extruded type cell printing, or described
Macroscopic hydrogel silk is cultivated to be formed after cell cluster and be carried out again.In described two different print procedures, the former is in building micro-loop
The characteristic of increment, migration and the self assembly using cell is needed during border, then direct construction goes out and internal similar cluster the latter
Shape tumor tissues.Two different methods, there are certain differences for obtained model, during can generating respectively to cell model
Cell growth etc. physiological activities and tumour medicine detection etc. carry out medical research.
In the present invention, the final cell number of the drip-type cell printing is single or multiple.Using the feature, in structure
It builds in foreign cell external model and proposes two kinds of techniques.First is that passing through the cell growth in extruded type cell printing water-setting collodion silk
Proliferation forms cell cluster, and drippage printing individual cells form microenvironment around it；Another kind is by extruded type cell
Cell in stamping ink gelatine silk is uniformly distributed to form microenvironment cell, and by drip printing added into water-setting collodion silk it is larger
The cell cluster of cell density, the larger cell concentration are 106~107；It can be applied to different demands for both techniques
In vitro cell model building in.
Cell kind in the present invention, in the cell category of the drip-type cell printing and the macroscopic hydrogel silk structure
Class is identical, or is the one or more cells different from cell category in the macroscopic hydrogel silk structure.When using of the same race
When cell, the change of local cell density may be implemented by drip-type cell printing method, different cell densities can be studied
Influence to cellular morphology, Reproduction methods and self assembly mode.It, can be between different cell categories when using various kinds of cell
Interaction is studied, and the artificial substituent similar with internal various kinds of cell can also be constructed.
In the present invention, the top certain altitude that the glass micro-nozzle can be suspended from the body support frame containing eucaryotic cell structure carries out specific
The drip-type cell printing of pattern or shape can also carry out drip-type cell printing at the grid-gap of bracket, to realize
Specific quantity cell is planted in bracket specific position.The drippage in the droplet of different positions may be implemented in this method, guarantees water
Variety classes cell distribution is controllable inside gelatine silk, guarantees spatial arrangement and information interchange between foreign cell, avoids
Foreign cell model caused by as the separation of cell spaces position fails.
(3) beneficial effect
1) present invention comprehensively utilizes micro- extruded type cell printing technology and drip-type cell precise Printing technology is integrated
The precisely controlled assembling of formula foreign cell, maximum feature are the printing of macroscopical cell cluster not only to have may be implemented, but also may be implemented
Cell cluster specific position carries out the fixed point printing of microcosmic individual cells, and the individual cells of the cell cluster and printing printed
It can be any two or a variety of heterogenous cells；The present invention is thin in cell solution by increasing during drip-type prints
The density of born of the same parents can directly carry out the printing of cluster cell, avoid to form the incubation that cluster needs to carry out.It is a kind of building
New process in foreign cell model method.
2) extruded type cell printing uses two step cross-linking methods in the technology, that is, includes first step physical crosslinking and second step
Learn crosslinking；Physical crosslinking, which refers to, crosslinks the components of gelatin in material by low temperature, becomes gel-like state；Chemical crosslinking is
Referring to is crosslinked the seaweed acid constituents in material by calcium ion.Two steps are crosslinked micro- extruded type printing technique and can efficiently construct
Highdensity cell/hydrogel material structural body, obtains 100~800 microns of water-setting collodion silk structure, and research finds that cell exists
In 100-800 microns of water-setting collodion silks by the integrated three-dimensional cluster-shaped eucaryotic cell structure of culture post-concentration, very high level simulates body
The form of inner cell assembling and aggregation, has important bionical value；
3) in the present invention and drip-type cell precise Printing using the cell printing skill driven based on the stagnant inertia force of alternation
Art, the technology can be accurately controlled the printing and assembling of individual cells by the alternately viscous force of variation and inertia force effect,
Realize using individual cells as minimum unit cell model construct, and the technique do not generated in print procedure high temperature, high pressure,
Strong electrical field and fluid salivate phenomenon, and survival rate is very suitable to cell printing technique 90% or more after cell printing.In short, this
Invention is integrated with two steps and had been crosslinked micro- extruded type printing technique and drip-type cell printing technology based on alternation stagnant inertia force driving both
It ensure that the efficient building of macroscopical cell cluster, guaranteed fixed point printings of the microcosmic individual cells on cluster cell have
Important pathology, pharmacology application potential.
Detailed description of the invention
Fig. 1 is foreign cell 3 D-printing system structure diagram
Fig. 2 is foreign cell 3 D-printing system nozzle structure enlarged diagram
Fig. 3 is that drip-type cell printing schematic diagram is carried out on just printing the hydrogel scaffold containing cell finished；
Fig. 4 is that drip-type cell printing spray head is suspended in outside hydrogel scaffold, and cell cluster is specific in hydrogel scaffold
Drip-type cell printing is carried out on position.
Fig. 5 is that drip-type cell printing spray head is inserted into hydrogel scaffold, and cell cluster is specific in hydrogel scaffold
Drip-type cell printing is carried out on position.
Fig. 6 is that multiple cell drip-type printings are carried out on the hydrogel scaffold of extruded type cell printing, and printing obtains thin
Born of the same parents' cluster schematic diagram.
In figure: 1, equipment molding room, 2, Z axis motion platform, 3, spray head clamping device, 4, piezoelectric ceramics, 5, horizontal observation
Microscope, 6, XY axis motion platform, 7, drip-type cell printing mini sprinkler, 8, cell/hydrogel material structural body, 9, culture
Ware, 10, injector drive motor, 11, heat preservation sleeve, 12,1mL syringe and spray head, 13, semiconductor chilling plate, 14, ultraviolet go out
Bacterium lamp, 15, Vertical Observation microscope, 16, signal generator, 17, power amplifier, 18, PMAC controller, 19, industrial personal computer, 20
Fine position motor, 21 pressure heads, 22 screw rods.
Specific embodiment
The following examples are used to illustrate the present invention, but are not intended to limit the scope of the present invention..
Embodiment 1
The present embodiment is related to a kind of foreign cell three-dimensional cell print system, including motion platform and is fixed on the movement
Extruded type cell printing mechanism and drip-type cell printing mechanism on platform, for observing the observing unit of cell printing position
And control unit；
Described control unit first controls the printing that extruded type cell printing mechanism completes cell hydrogel structure, then controls
Drip-type cell printing structure completes the printing of individual cells on the cell hydrogel structure；During printing, control
Unit control motion platform processed is mobile in tri- directions X, Y, Z, to realize extruded type cell printing mechanism and the drippage
Printing of the formula cell printing mechanism in different location.
Embodiment 2
The present embodiment is related to a kind of foreign cell 3 D-printing system, and structure is as depicted in figs. 1 and 2 the same as 1 phase of embodiment
Than difference is that the print system is placed in molding room 1, and ultraviolet sterilization lamp 14 is equipped in the molding room；
Extruded type cell printing mechanism is fixed on the Z axis motion platform 2 of motion platform, by injector drive motor
10, sleeve 11 is kept the temperature, 1mL syringe and its spray head 12 form；Drip-type cell printing mechanism by fine position motor 20,
Pressure head 21, piezoelectric ceramics 4 and drip-type cell printing are formed with mini sprinkler 7, are fixed on the Z by spray head clamping device 3
On axis motion platform 2；
The observing unit includes the horizontal microscope for observing 5 on the spray head clamping device 3 and is set to the molding room 1
The Vertical Observation microscope 15 of bottom；
Described control unit includes signal amplifier 16, power amplifier 17, PMAC controller 18 and industrial personal computer 19；Its
In, the industrial personal computer 19 is computer, computer and the PMAC controller 18 and the horizontal microscope for observing 5, described perpendicular
Straight microscope 15 is connected.Movement position adjusting on, the PMAC controller 18 respectively with the XY axis motion platform 6, the Z
Axis motion platform 2 is connected, and controls the movement of each axis.In the control of extruded type cell printing, the PMAC controller 18 and institute
It states injector drive motor 10 to be connected, and the parameters such as its movement velocity is adjusted.In the control of drip-type cell printing, institute
PMAC controller 18 is stated to be connected with signal generator 16 and fine position motor 20.The PMAC controller 18 controls signal
Device 16 generates waveform, and the waveform that signal generator 16 generates is transported to the piezoelectric ceramics after the power amplifier 17 amplification
The generation of droplet is carried out on 4.
The cell carrier includes the semiconductor chilling plate 13 on the XY axis plane of movement, training placed on it
It supports ware 9 and is put in cell/hydrogel material structural body 8 of the culture dish.
Embodiment 3
The present embodiment be related to it is a kind of utilize foreign cell 3 D-printing system of the present invention carry out tumor models
The method of printing, which comprises the steps of:
1) it is formulated for the cell solution of extruded type cell printing.It will cultivated after medium centrifugal containing A549 cell
It suspends in base, reaches 4 × 106The cell density of/ml.Later by the A549 cell culture fluid, 4% sodium alginate soln
It is mixed with 20% gelatin solution according to the ratio that volume ratio is 1:1:1, and makes final cell density 1 × 106/ ml is left
It is right.
2) it is configured to the cell solution of drip-type cell printing.By the A549 cell culture fluid carry out draw and it is dilute
It releases, so that final cell density is 5 × 104/ml.Wherein, it is worth noting that, the cell density in cell solution is not according to
Same model needs can be adjusted, and be not only limited to used cell density.
3) 15~30min of ultraviolet sterilization lamp is opened, sterilization treatment is carried out to the cell printing system；
4) it extruded type cell printing cell solution and drip-type cell printing is used for is filled described with cell solution respectively
Enter in the syringe and in micro-nozzle.
5) start described control unit and observing unit, in conjunction with the macromodel of design, progress extruded type cell first is beaten
Print.Model information becomes code by the compiling of the industrial personal computer and is sent to the PMAC controller, and the PMAC controller is then
According to obtained code, control includes the fortune of XY axis motion platform, the movement of Z axis motion platform and injector drive motor
It is dynamic.Mutual cooperation and linkage between above-mentioned motor shaft just realize the extrusion and printing of water-setting collodion silk.Meanwhile in observing unit
The case where water-setting collodion silk that horizontal microscope for observing and vertical microscope for observing can obtain printing, feeds back, from industrial personal computer
Display port on can obtain the relevant informations such as the forming quality of hydrogel.
6) start described control unit and observing unit, binding model design and real-time observed result, it is thin to carry out drip-type
Born of the same parents' printing.Drippage print position is judged according to practical cell distribution in model and water-setting collodion silk is pre-designed.Observing unit
The specific location that middle horizontal microscope for observing and vertical microscope for observing pass through cell in the available water-setting collodion silk of optical observation
Etc. relevant informations, after obtaining relevant information from the display port of industrial personal computer, by PMAC controller input code instruct wrap
Include the movement of XY axis motion platform, Z axis motion platform and the fine position motor so that the position of micro-nozzle nozzle be in
The top of dripping position.Then, it is instructed by industrial personal computer to the PMAC controller output code, the PMAC controller control
Signal generator exports pre-set waveform to the power amplifier, and the power amplifier drives after amplifying signal
The vibration of piezoelectric ceramics.After piezoelectric ceramic vibration is primary, primary formation and the drippage of drop are just realized.
7) step 6), available heterogeneous tumor models are repeated.
Printing type described in the present embodiment is progress drip-type printing immediately after the completion of extruded type printing, and schematic diagram is as schemed
3。
Embodiment 4
Compared with embodiment 3, the present embodiment carries out the printing of tumor models, and in the step 6), drip-type is thin
Born of the same parents' drip-type cell printing spray head is suspended in outside hydrogel scaffold, is dripped on the specific position of cell cluster in hydrogel scaffold
Fall formula cell drip-type cell printing, schematic diagram such as Fig. 4.The foreign cell model printed by this method, in incubation
Using the increment, movement and self-assembling technique of cell, during the cell model of building can the vital movement to cell can carry out
Test has potential application on carrying out tumor cell migration and tumor pathogenesis research.
Embodiment 5
Compared with embodiment 3, the present embodiment carries out the printing of tumor models, and in the step 6), drip-type is thin
Born of the same parents' drip-type cell printing spray head protrudes into hydrogel scaffold, is dripped on the specific position of cell cluster in hydrogel scaffold
Fall formula cell drip-type cell printing, schematic diagram such as Fig. 5.In the foreign cell model printed by this method, foreign cell
The distance between it is closer, the interaction between cell is more direct, building of the cell model of building to Vitro Tumor model, right
There is application in the influence of cell-cell interaction and tumour medicine detection.
Embodiment 6
Compared with embodiment 3, the present embodiment carries out the printing of tumor models, in the step 2), adjusts drippage
The cell concentration of formula printing is to 4 × 106/ ml can be constructed after improving cell density by the method for drip-type cell printing
Cell cluster.With embodiment 3) in method compared with, avoid to form cluster and need the incubation that carries out.It is that a kind of building is different
New process in cell plastid model method.It prints schematic diagram such as Fig. 6.
Although above the present invention is described in detail with a general description of the specific embodiments,
On the basis of the present invention, it can be made some modifications or improvements, this will be apparent to those skilled in the art.Cause
This, these modifications or improvements, fall within the scope of the claimed invention without departing from theon the basis of the spirit of the present invention.
Claims (8)
1. a kind of foreign cell 3 D-printing system, which is characterized in that including motion platform, be fixed on the motion platform
Extruded type cell printing mechanism and drip-type cell printing mechanism, for observe and feed back the observing unit of cell printing situation with
And control unit；
Described control unit controls the printing that extruded type cell printing mechanism completes three-dimensional cell hydrogel structure first, then controls
Drip-type cell printing processed mechanism completes the printing of single or multiple cells on the three-dimensional cell hydrogel structure；It is printing
During, described control unit controls the three-dimensional motion of the motion platform, to realize the printing in required position；
Described control unit includes industrial personal computer, PMAC controller, signal generator and power amplifier；
The industrial personal computer is connected with the PMAC controller and the observing unit, for receiving feedback information and controlling to PMAC
Device processed issues instruction；The three-dimensional motion of the PMAC controller control motion platform and squeezing for the extruded type cell printing machine
Out；Further, the PMAC controller also adjusts the position of drip-type cell printing mechanism, by control signal generator and
Power amplifier prints to control the drip-type of cell；
Drip-type cell printing mechanism includes the fine position motor being fixed on the Z axis of the motion platform, pressure head, past
Multiple movement mechanism and the drip-type cell printing mini sprinkler being attached thereto；Wherein, the mini sprinkler is fixed on reciprocating machine
On structure, the reciprocating mechanism is successively connected with pressure head and fine position motor；The PMAC controller and fine position electricity
Machine is connected, for finely tuning the position of the mini sprinkler；The PMAC controller also control signal generator generates waveform, the wave
Shape is transported on the reciprocating mechanism after power amplifier amplification, realizes drop by the movement of reciprocating mechanism
Fall formula printing.
2. print system according to claim 1, which is characterized in that extruded type cell printing mechanism includes being fixed on
Injector drive motor, pressure head, syringe and its spray head and heat preservation sleeve on the Z axis of the motion platform, wherein the spray
Head is mounted on syringe oral area, and syringe is fixed in heat preservation sleeve；Control of the injector drive motor in the PMAC controller
Under system, the continuous extrusion for realizing internal liquid is pushed syringe piston by the pressure head.
3. print system according to claim 1 or 2, which is characterized in that the observing unit includes that horizontal observation is micro-
Mirror and vertical microscope for observing.
4. the method for carrying out foreign cell 3 D-printing using any one of claims 1 to 33 print system, feature exist
In including the following steps:
1) sterilization treatment is carried out to the foreign cell 3 D-printing system；
2) according to the model being pre-designed, described control unit controls Z axis height and the position XY of the motion platform, and controls
Extruded type cell printing mechanism is printed, and three-dimensional cell hydrogel structure is obtained；
3) according to the information for model and the observing unit feedback being pre-designed or according only to the model being pre-designed, the control is single
Member adjusts Z axis height and the position XY of the motion platform, and by the fine tuning to drip-type cell printing mechanism position, realizes
The position of double spray head working condition switchings and drip-type cell printing positions；
4) described control unit controls drip-type cell printing mechanism and passes through the cell printing based on alternation stagnant inertia force driving
Technology carries out drip-type printing on the three-dimensional cell hydrogel structure, obtains foreign cell threedimensional model；
5) it is needed for different model, step 3) and 4) is repeated, until constructing required foreign cell threedimensional model.
5. according to the method described in claim 4, it is characterized in that, the three-dimensional cell water-setting of the extruded type cell printing
The condensation of glue is using physical crosslinking and is chemically crosslinked, and in the physical crosslinking, makes the three-dimensional cell hydrogel knot by low temperature
Components of gelatin in structure crosslinks, and becomes gel-like state；In the chemical crosslinking, the three-dimensional cellular water is made by calcium ion
Seaweed acid constituents in gel structure is crosslinked；Specifically, during the chemical crosslinking, the solid that is obtained to printing
2~4% CaCl is added in cell hydrogel structure2Solution extremely submerges, and keeps 2~4min in room conditions, then inhales
Solution is walked, just realizes the chemical crosslinking of the three-dimensional cell hydrogel structure, the diameter of water-setting collodion silk exists after crosslinking
Between 100-800 microns.
6. method according to claim 4 or 5, which is characterized in that the cell category of the drip-type cell printing and institute
The cell category stated in three-dimensional cell hydrogel structure is identical, or is and cell category in the three-dimensional cell hydrogel structure
Different one or more cells.
7. method according to claim 4 or 5, which is characterized in that the final cell number of the drip-type cell printing
It is single or multiple；Specifically, being single, operating procedure for final cell number are as follows: complete by extruded type cell printing
Cell cluster is formed at the growth and proliferation of cell in rear water-setting collodion silk, then the cluster cell peripheral drippage printing individual cells
Form microenvironment；It is multiple, operating procedure for final cell number are as follows: by extruded type cell printing water-setting collodion silk
Cell is uniformly distributed to form microenvironment, and the cell that larger cell density is directly added into water-setting collodion silk is printed by drip-type
Cluster.
8. method according to claim 4 or 5, which is characterized in that glass micro-nozzle can be suspended from the branch of body containing eucaryotic cell structure
The top certain altitude of frame carries out the drip-type cell printing of specific pattern or shape, or is dripped at the grid-gap of bracket
Formula cell printing is fallen, plants specific quantity cell in bracket specific position to realize.
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| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510455765.9A CN106399078B (en) | 2015-07-29 | 2015-07-29 | A kind of foreign cell 3 D-printing system and method |
Applications Claiming Priority (1)
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| CN107320773B (en) * | 2017-06-09 | 2020-08-18 | 西安交通大学 | Artificial muscle scaffold model and preparation device and method thereof |
| CN107599396B (en) * | 2017-11-10 | 2023-09-12 | 衡阳师范学院 | 3D printer shower nozzle unit and 3D printer |
| CN110118863A (en) * | 2018-09-21 | 2019-08-13 | 苏州永沁泉智能设备有限公司 | A kind of high-throughput drug automatic splinter screening device for screening and method |
| CN109897195A (en) * | 2019-02-15 | 2019-06-18 | 清华大学 | A kind of preparation method of 3D printing degradable microsphere and its application in the external micro-assembly robot of constructing function |
| CN109849329B (en) * | 2019-02-22 | 2021-10-29 | 河南科技大学 | Rapid forming method of magnetorheological complex |
| CN110229740A (en) * | 2019-06-14 | 2019-09-13 | 科先医疗科技(苏州)有限公司 | A kind of Biotype artificial liver cell 3D printing system |
| CN110669653B (en) * | 2019-09-24 | 2023-05-16 | 山东大学 | Single-cell automatic printing system |
| CN110680562B (en) * | 2019-10-25 | 2023-12-15 | 上海交通大学医学院附属第九人民医院 | Bone defect repair stent, construction method, preparation method, computer readable storage medium and equipment |
| CN110938585B (en) * | 2019-12-10 | 2021-11-12 | 清华大学 | Vascularized tissue construction method based on cell cluster 3D printing and application thereof |
| CN112776325B (en) * | 2020-12-29 | 2022-10-11 | 杭州电子科技大学 | Three-dimensional ultrasonic array support-free cell printing device and printing process thereof |
| CN113352603B (en) * | 2021-05-28 | 2023-04-14 | 西安交通大学 | Ultrasonic-assisted extrusion type cell printing nozzle and printing device |
| CN113524674B (en) * | 2021-07-07 | 2022-12-13 | 清华大学深圳国际研究生院 | Device and method for preparing three-dimensional patterned multi-material hydrogel heterostructure |
| CN113601834B (en) * | 2021-08-16 | 2023-06-23 | 杭州捷诺飞生物科技股份有限公司 | Three-dimensional forming method and system |
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