CN105524830A - Pretreatment liquid manufacturing structure of tissue engineering micromodule - Google Patents
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Abstract
The invention relates to the technical field of manufacturing of tissue engineering micromodules, and in particular relates to a pretreatment liquid manufacturing structure of a tissue engineering micromodule. The pretreatment liquid manufacturing structure comprises a first channel, a second channel and a third channel, wherein the first channel is used for conveying a cell solution; the second channel is used for conveying sheath liquid; the first channel and the second channel converge at one place, and pretreatment liquid formed in a manner that the sheath liquid wraps the cell solution is formed at the junction, and flows out from the third channel. The invention aims at proving the pretreatment liquid manufacturing structure of the tissue engineering micromodule, wherein the pretreatment liquid is liquid characterized in that the sheath fluid as the outer-ring laminar flow wraps the cell solution as the central laminar flow.
Description
Technical field
The present invention relates to organizational project micromodule field, particularly a kind of pretreatment fluid manufacturing structure of organizational project micromodule.
Background technology
Current organization engineering micromodule is faced with large, the active problem such as low of loss cell, particularly 3D biometric print technical field wherein.
Printing based on micromodule is the emerging method of of organizational project, this method injects cell in the inside of gel (as sodium alginate) drop, again gel drop is cured as a micromodule, such shell/core (gel/cell) structure avoids the impact that inner active cells is subject to outside atmosphere, substantially increase activity and the stability of cell in biometric print process, the mode of this micromodule is also widely used in cell cultures, drug delivery and release etc.
Traditional manufacture method, as spray method, has very large damage to cell, and can not ensure the shape and size of micromodule, more can not control shell/core Thickness Ratio.
On the other hand, micro-fluidic chip is also referred to as chip lab.It utilizes the feature such as laminar flow and low reynolds number of fluid under micro-meter scale, and to the material in fluid, as cell, particle, or chemical example etc. carries out synthesizing, is separated, wraps up, cultivates, detects and the operation such as screening.Utilize the feature of two-phase incompatibility, in micro-fluidic chip, microlayer model can be manufactured high-throughput.Its principle is: two kinds of immiscible liquid (or liquids and gases) are injected microchannel simultaneously, and wherein a kind of is external phase (being generally oil phase or gas), and another kind is disperse phase (cell solution).When two-phase is met, under shearing force and capillary effect, disperse phase fragments into microlayer model.
At present, main employing microfluidic chip structure that is T-shaped or flow focusing type produces microlayer model, by the trend of chip channel, and the adjustment of two phase pressures, the size and dimension of microlayer model can be controlled accurately.
But the microlayer model that these two kinds of structures produce, cell and gel mix, and namely cell is evenly distributed in gel, and the micromodule after final solidification is the spheroid of many entities, and is not the shell/nuclear structure satisfied the demands.The cell on micromodule surface, then can be dead because of contact oil phase.The cell of micromodule inside, is separated with gel between cell and cell, and cell is isolated in a series of little space, can not Effective multiplication and migrating.And continuous print oil phase can produce irreversible damage to cytoactive, must allow cell get along well as much as possible oil phase contact.Therefore, in order to meet the demand of 3D biometric print, cell cultures, drug delivery and release, a kind of method and the chip that realize shell/nuclear structure micromodule reliably, easily, is efficiently needed.
And the structure of the application, relate to the core texture in this new chip.This structure may be used for generating a kind of pretreatment fluid, more this pretreatment fluid is formed microlayer model by aforesaid method or additive method, is solidified into organizational project micromodule further.
Summary of the invention
The object of the invention is to overcome above-mentioned deficiency existing in prior art, provide a kind of manufacturing structure producing the pretreatment fluid of organizational project micromodule, described pretreatment fluid is the liquid that laminar flow, sheath fluid are outer ring laminar flow centered by cell solution.
In order to realize foregoing invention object, the invention provides following technical scheme:
A pretreatment fluid manufacturing structure for organizational project micromodule, it comprises:
First channel, for carrying cell solution;
Second passage, for carrying sheath fluid;
Third channel;
Described first channel, second passage cross at a place, form the described pretreatment fluid being wrapped in cell solution outside with described sheath fluid, flow out from third channel in intersection.
By the mode that the first channel of the application, second passage, third channel coordinate, can form a kind of pretreatment fluid, its internal flow is cell solution, and the fluid of outer layer covers is sheath fluid.Then this fluid is again by the method described in background technology, and described pretreatment fluid is formed the microlayer model of single dispersing phase, final curing is organizational project micromodule.
The micromodule that our scheme is formed is the described shell/nuclear structure meeting organizational project demand.Inside is cell solution, and the outside gel housing for solidification, give good protection to cell.Compare the mode of direct mixing of the prior art, cytoactive is very high, is leaping of a matter.
As preferred version of the present invention, described intersection is a cavity, described first channel is surrounded by inside cavity space at the entrance at described cavity place, control sheath fluid and first flow into cavity, after filling up cavity, cell solution enters again, because entrance is surrounded by inside cavity space, just naturally form the structure of parcel, then flowed out by third channel.
As preferred version of the present invention, the bore of described third channel and described cavity connectivity part is greater than the inlet calibre of described first channel at described cavity place, it can make the pretreatment fluid of formation be compressed, not easily turbulization, and the laminar flow of this structure is more stable.
As preferred version of the present invention, described first channel is divided into first paragraph passage and second segment passage, described first paragraph passage is permanent footpath section, and axis is straight line, described second segment passage is provided with bend, and one end of described first paragraph passage connects described intersection, the other end of described first paragraph passage is connected with described second segment passage, make the cell solution entering described intersection more stable, be convenient to the structure forming described pretreatment fluid, and the pretreatment fluid flowed out is also more stable.
As preferred version of the present invention, described second passage comprises first paragraph passage and second segment passage; Described first paragraph passage is permanent footpath section, and axis is straight line; Described second segment passage is provided with bend; One end of described first paragraph passage connects described intersection, and the other end of first paragraph passage is connected with described second segment passage, makes the described sheath fluid of inflow intersection also more stable, also convenient adjustment pressure, and the pretreatment fluid structure of formation is more perfect.
As preferred version of the present invention, the dead in line of the first paragraph passage on the axis of the first paragraph passage on described first channel and described third channel, makes sample liquid be tending towards the center of sheath fluid, forms coaxial pretreatment fluid.
As preferred version of the present invention, first paragraph passage angle on the first paragraph passage of described second passage and described first channel is greater than 0 °, is less than or equal to 90 °, reduces sheath fluid and flows into the resistance brought, prevent the adverse current of cell solution, also make the ectonexine structure of pretreatment fluid more stable simultaneously.
As preferred version of the present invention, described first channel, second passage, third channel be permanent footpath passage, namely the cross-sectional area of single passage remains unchanged, and facilitates pressure-controlling, and the flow rate of liquid simultaneously in each passage is also more stable.
As preferred version of the present invention, described second passage quantity is equal to or greater than two, be distributed in the both sides of the first paragraph passage of described first channel, continue to pour into sheath fluid from the both sides of cavity, more easily form the ectonexine structure of pretreatment fluid, and symmetrical configuration, facilitate pressure source control pressure, make the formation better effects if of pretreatment fluid.
As preferred version of the present invention, described second passage is even number, and is symmetrically distributed in the both sides of described first channel, when pouring into sheath fluid from cavity both sides, because the second passage quantity of both sides is identical, so flow more easy to control, pressure and other parameters.
As preferred version of the present invention, the cross section of described first channel, second passage, third channel is rectangle, and convenient processing, such as carries out photoetching process.
As preferred version of the present invention, manufacturing structure is formed by two chip block bondings, independent chip is provided with corresponding described first channel, second passage, the combination slot of third channel, after two pieces of described chip bondings, described combination slot coordinates the described first channel of formation, second passage, third channel, and at described first channel, the intersection of second passage and third channel forms cavity, because this manufacturing structure is a very little structure, so adopt the mode of bonding to process, can be simple a lot, processing is fast, cost is lower, separately two portions, independent chip carries out combination slot described in photoetching by photoetching technique.
As preferred version of the present invention, described first channel, second passage, third channel divide the axis of other first paragraph passage to be all positioned at same plane, when making the present invention form passage by two chip block bondings, reduce the difficulty of photoetching, more easily obtain the structure of sheath fluid parcel sample liquid simultaneously.
As preferred version of the present invention, the cross section of described first channel, second passage, third channel is circle.
As preferred version of the present invention, described second passage and described first channel are greater than 0 ° at the angle of described intersection, be less than or equal to 90 ° of modes being compared to right angle and arranging, more easily produce the structure of the pretreatment fluid of stable inside and outside laminar flow, and prevent the generation of adverse current, turbulent flow, also pressure more easy to control.
As preferred version of the present invention, the sheath fluid of described second passage conveying is gelating soln, and the oil phase of convenient and follow-up calcium ions produces solidification.
As preferred version of the present invention, the pretreatment fluid manufacturing structure of organizational project micromodule is formed by two chip block bondings, independent chip is provided with the combination slot of corresponding described first channel, second passage, third channel, after two pieces of described chip bondings, described combination slot coordinates formation described first channel, second passage, third channel, and forms cavity in the intersection of described first channel, second passage and third channel.
As preferred version of the present invention, described combination slot is rectangular tank, and on single described chip, the rectangular tank degree of depth of corresponding described first channel is less than the rectangular tank degree of depth of corresponding described cavity, after such bonding, the entrance of first channel at cavity place just wrap up by the internal space of cavity.
As preferred version of the present invention, the rectangular tank degree of depth of corresponding described third channel is identical with the rectangular tank degree of depth of the described cavity of correspondence, provides output environment, cavity inner wall and the transition of third channel interior walls be smooth that one stable to pretreatment fluid after being formed as far as possible.
As preferred version of the present invention, the outside Bonding pressure source of described first channel, second passage, described pressure source pressure adjustable, adaptability is stronger, by regulating pressure, the final described shell/nuclear structure forming different thickness ratio.
compared with prior art, beneficial effect of the present invention:
This structure can produce the pretreatment fluid that laminar flow, sheath fluid are outer ring laminar flow centered by cell solution, increases the cytoactive of final micromodule and the activity of follow-up generation tissue.
accompanying drawing illustrates:
Fig. 1 is the embodiment of the present invention 1 structural representation;
Fig. 2 is the embodiment of the present invention 4 structural representation;
Fig. 3 is principle of the invention figure;
Fig. 4 is the structural representation of second segment passage band bend in the embodiment of the present invention 1;
Fig. 5 is the pretreatment fluid cross sectional representation limited in the embodiment of the present invention 3;
Fig. 6 is application flow schematic diagram of the present invention;
Fig. 7 is application structure schematic diagram of the present invention;
Fig. 8 is cavity place structural representation in the embodiment of the present invention 1;
Fig. 9 is cavity place structural representation in the embodiment of the present invention 2;
Figure 10 is cavity place structural representation in the embodiment of the present invention 3;
Mark in figure: 13-second passage, 14-first channel, 23-third channel, 11-first inlet, 12-second inlet, 21-the 3rd inlet, 41-delivery port, 111-first paragraph passage, 112-second segment passage, 51-cavity.
Embodiment
Below in conjunction with embodiment and embodiment, the present invention is described in further detail.But this should be interpreted as that the scope of the above-mentioned theme of the present invention is only limitted to following embodiment, all technology realized based on content of the present invention all belong to scope of the present invention.
embodiment 1
As Fig. 1, a kind of pretreatment fluid manufacturing structure of organizational project micromodule, it comprises:
First channel 14, for carrying cell solution;
Second passage 13, for carrying sheath fluid, described first channel 14, the pressure source (being generally syringe pump or vacuum pump, with peristaltic pump in few situation) that the outside Bonding pressure of second passage 13 is adjustable;
Third channel 23;
As Fig. 3, 8, described first channel 14, second passage 13 crosses (third channel 23 also connects this intersection) at a place, the described pretreatment fluid being wrapped in cell solution outside with described sheath fluid is formed in intersection, flow out from third channel 23, described intersection is a cavity 51, by the encirclement of cavity 51 internal space, (namely the dotted portion in Fig. 8 is the cavity 51 described in us to the entrance of described first channel 14 at described cavity 51 place, can see in figure, in the present embodiment, cavity 51 is wrapped in the outlet of this place's first channel 14, in real work, sheath fluid is first injected into, after cavity is filled, cell solution is injected into again, in the present embodiment, because cavity 51 is outlets of being wrapped in this place's first channel 14, so cell solution has just been wrapped up by sheath fluid after injecting naturally very much, the pretreatment fluid formed also can flow out from third channel 23, do not need to be compressed, the laminar flow structure formed is more stable), described third channel 23 is greater than the inlet calibre of described first channel 14 at described cavity 51 place at the inlet calibre at described cavity 51 place, the dead in line of the first paragraph passage 111 on the axis of the first paragraph passage 111 on described first channel 14 and described third channel 23, described first channel 14, second passage 13, third channel 23 be permanent footpath passage, namely the cross-sectional area of single passage remains unchanged, described first channel 14, second passage 13, the axis of third channel 23 points of other first paragraph passages 111 is all positioned at same plane.
As Fig. 7, described first channel 14, second passage 13 are divided into first paragraph passage 111 and second segment passage 112, described first paragraph passage 111 is permanent footpath section, and axis is straight line, second passage 112 is provided with bend, and one end of the first paragraph passage 111 of described first channel 14/ second passage 13 connects described cavity respectively; The second segment passage 112 of corresponding described first channel 14/ second passage 13 that is connected of the other end difference of the first paragraph passage 111 of described first channel 14/ second passage 13.
In the present embodiment, the first paragraph passage 111 of described second passage 13 is greater than 0 ° with first paragraph passage 111 angle on described first channel 14, and being less than 90 °, is acute angle, as Fig. 1.
As Fig. 1,6,7, described second passage 13 quantity is equal to or greater than two (in the present embodiment being two), is distributed in the both sides of the first paragraph passage 111 of described first channel 14 symmetrically.
As Fig. 5, described first channel 14, second passage 13, the cross section of third channel 23 is rectangle, form (as Fig. 7 by two chip block bondings, schematic diagram during embody rule), independent chip is provided with corresponding described first channel 14, second passage 13, third channel 23, the rectangular tank (i.e. aforesaid combination slot) of cavity 51, after two pieces of described chip bondings, described rectangular tank coordinates the described first channel 14 of formation, second passage 13, third channel 23, cavity 51, on single described chip, the rectangular tank degree of depth of corresponding described first channel 14 is less than the rectangular tank degree of depth of corresponding described cavity 51.
As Fig. 4,7, the second segment passage 112 of described first channel 14, second passage 13 is provided with bend, the second segment passage 112 of described third channel 23 is permanent footpath section, and first paragraph passage 111 dead in line of axis and third channel 23.
In the present embodiment, the sheath fluid that described second passage 13 is carried is gelating soln, concrete, adopts sodium alginate soln.
embodiment 2
As Fig. 9, namely dotted portion is the cavity 51 described in us, can see in figure, in the present embodiment, the outlet of cavity 51 and first channel 14 seamlessly transits, in real work, sheath fluid is first injected into, after cavity is filled, cell solution is injected into again, in the present embodiment, because the outlet of cavity 51 and first channel 14 seamlessly transits, accordingly, the bore of third channel 23 and the bore of first channel 14 are identical, so cell solution can be obtained more carefully a bit by sheath hydraulic compression after injecting, pretreatment fluid could be wrapped to form by sheath fluid in cavity, the pretreatment fluid formed flows out from third channel 23, compare the structure in embodiment 1, the laminar flow structure of pretreatment fluid need could keep stable after for some time adjustment, form thinner pretreatment fluid structure, meet the different needs.
embodiment 3
As the cross sectional representation of first in Fig. 5, in the present embodiment, except the structure identical with in embodiment 1, the cross section of described first channel 14, second passage 13, third channel 23 is circle;
The cross section of described first channel 14, second passage 13, third channel 23 not necessarily adopts the same shape, it can be difform combination, such as Fig. 5, difform combination still can form the structure of the pretreatment fluid of sodium alginate soln parcel cell solution, just cross section is different, and Fig. 5 is followed successively by:
Third channel 23 cross section is square, and first channel 14 cross section is square;
Third channel 23 cross section is square, and first channel 14 cross section is circular;
Third channel 23 cross section is circular, and first channel 14 cross section is square;
Third channel 23 cross section is circular, and first channel 14 cross section is circular.
These structures still can meet the generation operation of follow-up microlayer model, and curing process, also can produce satisfactory micromodule, can carry out the selection in cross section, namely select on single chip, the shape of slotting according to process requirements;
In general, adopting photoetching process to process this fluting, is rectangular tank, but occurs eliminating after processing other circular modes, certainly, is also not limited only to circle disclosed in the present embodiment, the square or form that combines.
As Figure 10, namely dotted portion is the cavity 51 described in us, can see in figure, in the present embodiment, from the outlet of first channel 14, cavity 51 diminishes gradually, in real work, sheath fluid is first injected into, after cavity is filled, cell solution is injected into again, in the present embodiment, due to the outlet from first channel 14, cavity 51 diminishes gradually, accordingly, the bore of the relative aperture first channel 14 of third channel 23 is little, so meeting must than thin a lot of in the past by sheath hydraulic compression after cell solution injection, pretreatment fluid could be wrapped to form by sheath fluid in cavity, the pretreatment fluid formed flows out from third channel 23, compare the structure in embodiment 2, the laminar flow structure of pretreatment fluid need could keep stable after longer for some time adjustment, form thinner pretreatment fluid structure, meet the different needs, the micromodule that final formation is less.
Concrete application, as Fig. 6,7, after producing satisfactory pretreatment fluid, is extruded pretreatment fluid for a series of microlayer model by oil-continuous phase, just defines required micromodule after being solidified by calcium ion.
Realize the device of this process as Fig. 7, except the structure of the present invention itself (in figure A place), also have the part and other parts (in other partial graphs 7 not display) that form microlayer model, and whole chip after bonding has the first inlet 11, second inlet 12, 3rd inlet 21, delivery port 41, as Fig. 7, corresponding, first inlet 11, second inlet 12, 3rd inlet 21 injects sodium alginate soln respectively, cell solution, oil-continuous phase, final microlayer model exports from delivery port 41, enter curing process, or delivery port 41 is not set, increase new pipeline, curing process is realized in chip.
embodiment 4
As Fig. 2, in the present embodiment, except the structure identical with embodiment 1, the first paragraph passage 111 of described second passage 13 is right angle with first paragraph passage 111 angle on described first channel 14.
Claims (10)
1. a pretreatment fluid manufacturing structure for organizational project micromodule, is characterized in that, comprising:
First channel, for carrying cell solution;
Second passage, for carrying sheath fluid;
Third channel;
Described first channel, second passage cross at a place, form the pretreatment fluid being wrapped in cell solution outside with described sheath fluid, and described pretreatment fluid flow out in intersection from third channel.
2. the pretreatment fluid manufacturing structure of organizational project micromodule according to claim 1, is characterized in that, described intersection forms a cavity, and described first channel is surrounded by inside cavity space at the entrance at described cavity place.
3. the pretreatment fluid manufacturing structure of organizational project micromodule according to claim 2, is characterized in that, the bore of described third channel and described cavity connectivity part is greater than the inlet calibre of described first channel at described cavity place.
4. the pretreatment fluid manufacturing structure of organizational project micromodule according to claim 1, is characterized in that, described first channel comprises first paragraph passage and second segment passage; Described first paragraph passage is permanent footpath section, and axis is straight line; Described second segment passage is provided with bend; One end of described first paragraph passage connects described intersection, and the other end of first paragraph passage is connected with described second segment passage.
5. the pretreatment fluid manufacturing structure of organizational project micromodule according to claim 1, is characterized in that, described second passage comprises first paragraph passage and second segment passage; Described first paragraph passage is permanent footpath section, and axis is straight line; Described second segment passage is provided with bend; One end of described first paragraph passage connects described intersection, and the other end of first paragraph passage is connected with described second segment passage.
6. the pretreatment fluid manufacturing structure of organizational project micromodule according to claim 4, is characterized in that, the axis of the first paragraph passage on described first channel and the dead in line of described third channel.
7. the pretreatment fluid manufacturing structure of organizational project micromodule according to claim 1, is characterized in that, described first channel and second passage include first paragraph passage and second segment passage; And the first paragraph passage of described first paragraph passage and second segment passage is permanent footpath section, and axis is straight line; The second segment passage of described first paragraph passage and second segment passage is equipped with bend; One end of the first paragraph passage of described first channel/second passage connects described intersection respectively; The second segment passage of the corresponding described first channel/second passage that is connected of the other end difference of the first paragraph passage of described first channel/second passage; Angle between first paragraph passage on the first paragraph passage of described second passage and described first channel is greater than 0 °, is less than or equal to 90 °.
8. the pretreatment fluid manufacturing structure of organizational project micromodule according to claim 1, is characterized in that, described first channel, second passage, third channel be permanent footpath passage, namely the cross-sectional area of single passage remains unchanged.
9. the pretreatment fluid manufacturing structure of organizational project micromodule according to claim 1, is characterized in that, described second passage quantity is equal to or greater than two, and is distributed in the both sides of described first channel.
10. the pretreatment fluid manufacturing structure of organizational project micromodule according to claim 9, is characterized in that, described second passage is even number, and is symmetrically distributed in the both sides of described first channel.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109746061A (en) * | 2017-11-06 | 2019-05-14 | 北京新羿生物科技有限公司 | Microlayer model generating means |
| CN109735429A (en) * | 2019-01-28 | 2019-05-10 | 佛山市铬维科技有限公司 | Micro-fluidic chip and the system and its separation method for separating various kinds of cell |
| CN109735429B (en) * | 2019-01-28 | 2024-05-10 | 珠海银方舟生物芯片有限公司 | Microfluidic chip, system for separating multiple cells and separation method thereof |
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