CN106929417A - A kind of multi-layer cellular culture micro element bionical based on vein eyed structure - Google Patents
A kind of multi-layer cellular culture micro element bionical based on vein eyed structure Download PDFInfo
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Abstract
The invention provides a kind of multi-layer cellular culture micro element bionical based on vein eyed structure, belong to carries out micro element design field using bionical means.The multi-layer cellular culture micro element is made up of three-decker, respectively cell sample introduction layer, intermediate layer and bionical culture layer.Using transition zone connection cell sample introduction layer and bionical culture layer with through-hole structure.Microchannel simulation vein network structure in wherein bionical culture layer, cell culture insert simulation eyed structure is embedded at the bifurcation point of microchannel network passage, for connecting the pit structure on microtrabeculae interstitial structure simulation vein duct wall of the microchannel network with culture pond.Multi-layer cellular culture micro element of the invention has bio-compatibility higher, makes simple, low cost, it is possible to achieve produce in enormous quantities.
Description
Technical field
The invention belongs to carry out micro element design field using bionical means, and in particular to one kind is based on vein eyed structure
Bionical multi-layer cellular culture micro element, the cell culture micro element is used to build the homogeneous stream of cell injuring model dynamic stability
Body flows microenvironment.
Background technology
Cell is the elementary cell for constituting organism, and cell research is for disclosing life rule, diagnosing the illness, screen medicine
Etc. significant.Cell culture is the basis for carrying out cell research, traditional cell injuring model often using culture dish,
Blake bottle or culture plate, are a kind of static cultures, different in body flowing microenvironment from cell.
The microchannel network of the micro-fluidic device vascular system residing in body with cell is closely similar, can enter Mobile state
Culture, therefore using microflow control technique carry out cell culture make cell the in-vitro simulated of body microenvironment and control turn into can
Energy.
Culture pond is one of critical function unit of micro-current controlled cell culture device, to ensure each cell tool in cultivation region
There is identical microenvironment, it is desirable to which the flow field in cell culture area has homogeneity.Meanwhile, to ensure the repdocutbility of experiment, it is desirable to thin
The flow field of born of the same parents cultivation region has stability.Therefore key of the culture pond as micro element design how is designed.
The Nature forms many optimization structures by long-term evolution.Dicotyledon vein will can be transported to from root
The liquid of petiole is uniformly transported to each mesophyll cell, and supply mesophyll cell carries out every physiological activity.Dicotyledon leaf
This hydraulic characteristic(s) of arteries and veins is inseparable with structure.Dicotyledon vein has following feature:Vein is in integrally net distribution,
And in master pulse fluid flow change gradually tended towards stability with the raising of middle arteries and veins density, therefore flow change to mesophyll cell
Influence is small, it is uniform to be delivered to each region nutrient of blade, and highdensity closure vein can ensure that liquid transport will not be due to
Local vein damage or entrained gas and block, it is ensured that the stability of liquid supply in mesh region.
The content of the invention
It is an object of the present invention to overcome the shortcoming of prior art, flow of fluid micro-loop is influenceed in organism using extracting
The method of the micro-structural in border, design is a kind of can to build the thin of the homogeneous cell injuring model flow of fluid microenvironment of dynamic stability
Born of the same parents cultivate micro element, and the micro element is made and for cell injuring model using micro-processing technology.
Technical scheme:
A kind of multi-layer cellular culture micro element bionical based on vein eyed structure, including cell sample introduction layer 1, middle transition
Layer 2 and bionical culture layer 3, cell sample introduction layer 1 are located at the superiors, and intermediate layer 2 is located at intermediate layer, and bionical culture layer 3 is located at
Bottom;
Described cell sample introduction layer 1 is provided with 4 through holes and 1 alignment hole, 4 through holes be respectively nutrient solution sample holes 4,
Cell sample holes 5, cell sample outlet hole 8 and waste liquid tap 9, alignment hole 7 are located at the center of cell sample introduction layer 1, and nutrient solution enters
Sample hole 4 and waste liquid tap 9 are symmetrically positioned in alignment hole 7 or so and hold, and cell sample holes 5 and cell sample outlet hole 8 are symmetrically positioned in alignment
The upper and lower side of hole 7;Connected by passage 6 between cell sample holes 5 and cell sample outlet hole 8, passage 6 is arranged symmetrically in pairs;Go out with cell
The connected side of passage 6 in sample hole 8 is transversely provided with trapezoidal micro- dam 18, for stopping that cell is rushed out culture pond;
Described intermediate layer 2 is provided with 3 through holes, is from left to right followed successively by nutrient solution sample introduction through hole 10, culture pond
Intercommunicating pore 11 and waste liquid discharge through hole 12, nutrient solution sample introduction through hole 10 are communicated with the nutrient solution sample holes 4 of cell sample introduction layer 1, given up
Liquid is discharged through hole 12 and is communicated with the waste liquid tap 9 of cell sample introduction layer 1, and culture pond intercommunicating pore 11 aligns with cell sample introduction layer 1
Hole 7 is relative, and shape size is identical;
Described bionical culture layer 3 is provided with nutrient solution liquid storage tank 13, bionical culture pond 15 and waste liquid liquid storage tank 17, culture
Liquid enters nutrient solution liquid storage tank 13 from nutrient solution sample holes 4 by nutrient solution sample introduction through hole 10, nutrient solution liquid storage tank 13 with it is bionical
It is connected by nutrient solution sample intake passage 14 between culture pond 15, going out sample by waste liquid between bionical culture pond 15 and waste liquid liquid storage tank 17 leads to
Road 16 is connected;
13 points described of nutrient solution liquid storage tank is liquid storage area 19 and overflow area 20, and liquid storage area 19 is located at nutrient solution liquid storage tank 13
Penetralia, liquid storage area 19 surrounds by multiple fence microtrabeculaes 21, and the inscribed circle diameter of liquid storage area 19 is logical not less than nutrient solution sample introduction
The inscribed circle diameter in hole 10, nutrient solution enters overflow area 20 by fence microtrabeculae gap 22, and overflow area 20 leads to nutrient solution sample introduction
Road 14 is communicated;
Described bionical culture pond 15 is provided with two-layer microtrabeculae, is interior microtrabeculae layer and outer microtrabeculae layer, and interior microtrabeculae layer is by multiple
The polygonized structure that microtrabeculae 24 is surrounded in arrangement closely, the polygonized structure inside surrounded by interior microtrabeculae 24 is cell culture area
23, nutrient solution enters cell culture area 23 by the interior microtrabeculae gap 28 between interior microtrabeculae 24;It is outer that outer microtrabeculae layer is located at interior microtrabeculae layer
Side, the quantity of the outer microtrabeculae 25 arranged in outer microtrabeculae layer is identical with polygonal side number, by the passage inside bionical culture pond 15
It is divided into the one-level fork fraction trouble passage 27 of subchannel 26 and two;One-level fork subchannel 26 is bionical culture pond 15 and outer microtrabeculae
The passage formed between passage, outer microtrabeculae layer China and foreign countries microtrabeculae 25 that interlayer is formed, the hydraulic diameter of one-level fork subchannel 26 meets
The cube sum of Murray laws, i.e. one-level the fork hydraulic diameter of subchannel 26 is equal to the water conservancy diameter of nutrient solution sample intake passage 14
Cube;Two fraction trouble passages 27 are the passage that outer microtrabeculae layer and interior microtrabeculae interlayer are formed, and two fraction trouble passages 27 are
Afterbody subchannel, the most thin vein of simulation blade has identical hydraulic diameter with one-level fork subchannel 26.
Passage 6 above described cell sample introduction layer 1 is arranged symmetrically in pairs on alignment hole 7, makes cell from multiple directions
Into culture pond, uniformly it is distributed in cell culture area 23 beneficial to cell;Passage 6 above described cell sample introduction layer 1 with
Alignment hole 7 has identical depth;The height on the trapezoidal micro- dam 18 above described cell sample introduction layer 1 and the depth phase of passage 6
Together;Alignment hole 7 above described cell sample introduction floor 1 is of similar shape with culture pond intercommunicating pore 11, cell culture area 23
And size, it is easy to bonding to be aligned.
Vein structure of the described design inspiration of bionical culture layer 3 in dicotyledon blade.Vein ranking score
The design feature of trouble and closure increased the redundancy of moisture movement in blade, for the change of adjustment fluid flow is provided in vein
Mulitpath.Inspired by this phenomenon, the present invention devises the culture layer of imitative vein structure, mainly imitated at vein thready pulse
Structure.Nutrient solution sample intake passage 14, one-level fork subchannel 26 in the bionical culture layer 3, the mould of two fraction trouble passage 27
Intend vein network structure, with grade fork feature;The simulation eyed structure of bionical culture pond 15 in the bionical culture layer 3,
With polygon feature;The pit structure on interior microtrabeculae gap 28 simulation vein duct wall on the bionical culture pond, uses
In the fraction trouble passage 27 of connection two and cell culture area 23.
Described cell sample introduction layer 1, intermediate layer 2 and bionical culture layer 3 use thermosetting polymer poly dimethyl silicon
Oxygen alkane (PDMS), or thermoplastic polymer polymethyl methacrylate (PMMA) or polystyrene (PS) or makrolon (PC)
Material.
Described cell sample introduction layer 1, intermediate layer 2 and bionical culture layer 3 are made using micro fabrication, are made
Process is:Micro-mould is made using photoetching technique and dry etching technology, using Soft lithograph technology or hot-pressing technique by micro-mould
On pattern transfer to micro element material on.
The material thickness that described intermediate layer 2 is used is less than or equal to the height of the microtrabeculae on mould, to form through hole.
Described cell sample introduction layer 1 using micro-fluidic chip special-purpose punching device nutrient solution sample holes 4, cell sample holes 5,
Cell sample outlet hole 8 and the correspondence position of waste liquid tap 9 get through hole.
Described cell sample introduction layer 1, intermediate layer 2 and bionical culture layer 3 are processed or hot pressing by oxygen plasma
Method be bonded together.
In described bonding process, cell sample introduction layer 1 and intermediate layer 2 between need be directed at, intermediate layer 2 with imitate
Alignment is needed between raw culture layer 3, described alignment procedures use visual micro-imaging technique, alignment procedures are presented on
On computer screen, make to realize alignment between chip layer by the position between the upper lower platform of adjustment.
Described cell culture micro element is passed through absolute ethyl alcohol to keep in closed channel and culture pond after bonding is good
The hydrophily on surface, and play a part of sterilization.
Described cell culture micro element measures fluid in cell culture area 23 by the method that trace particle tests the speed
Flowing velocity.
Described cell culture micro element, silicone tube and pipe joint autoclave sterilization 1 before for cell culture are small
When, rat tail collagen protein or fibronectin are coated with after bacterium of having gone out, incubator is placed in overnight after being coated with collagen, beneficial to subsequent cell
It is adherent.
Described cell culture micro element stands a moment after cell is injected, and is placed in basis of microscopic observation cell injection feelings
Condition, is put into CO2Incubator static incubation is adherent to most cells.
Described cell culture micro element proceeds by dynamic cultivation after cell attachment.
Described cell culture micro element carried out one every 24 hours during dynamic cultivation to the cell in culture pond
Secondary observation is taken pictures to cell and is paved with whole culture pond.The quantity of each time period inner cell is counted, cell growth curve is drawn.
Beneficial effects of the present invention:Symmetrical cell sample intake passage is injected by the multidirectional cell of multichannel in pairs
Realize being uniformly distributed for culture pond inner cell;The bionical culture pond built by simulating plant vein eyed structure can be accurate
Control culture pond in flow of fluid microenvironment, analog cell body flow microenvironment, improve cells in vitro experimental result can
By property;Bionical culture pond has certain robustness, can resist external disturbance, reduces due to increasing Micropump, micro-valve, micro- logical
The destruction of road, concentration gradient generator or passage blocks the change of the culture pond flow field for causing, and improves the repeatability of experiment,
And the bionical culture pond is desirably integrated into various kinds of cell manipulation micro element, while cell culture microenvironment is ensured, is realized
The high-volume culture of cell;The micro element has bio-compatibility higher, makes simple, low cost, it is possible to achieve high-volume
Production.
Brief description of the drawings
Fig. 1 is the tomograph of cell culture micro element in the present invention.
Fig. 2 is the two-dimensional top-down view of cell sample introduction layer 1 in the present invention.
Fig. 3 is M partial enlarged drawings in Fig. 2, and cipollettic dam 18 is used to prevent cell from being rushed out culture pond.
Fig. 4 is that cell goes out sample area three-dimensional structure chart on cell sample introduction layer 1, and cipollettic dam 19 is located in passage 6, its highly with
The depth of passage 6 is identical.
Fig. 5 is the two-dimensional top-down view of bionical culture layer 3 in the present invention.
Fig. 6 is the three dimensional sectional view on A-A sections in Fig. 5, the figure shows nutrient solution liquid storage area 19, overflow area 20, training
Nutrient solution sample intake passage 14, cell culture area 23, waste liquid sample output passage 16 and waste liquid liquid storage tank 17 have identical depth.
Fig. 7 is the enlarged diagram of nutrient solution liquid storage tank 13 in Fig. 5.
Fig. 8 is the enlarged diagram of bionical culture pond in Fig. 5.
Fig. 9 is N partial enlarged drawings in Fig. 8, the pit structure on the interior simulation of microtrabeculae gap 28 vein catheter wall, is used to connect
Two fraction trouble passages 27 and cell culture area 23, cell culture area 23 is transported to by nutrient solution, and by cell culture area 23
Waste liquid derive.
Figure 10 is that the particle trajectory figure that trace particle tests the speed is carried out in artificial cell culture micro element of the present invention.
Figure 11 is the average speed of equally distributed 28 particles in the culture pond of artificial cell culture micro element of the present invention.
Figure 12 be in the culture pond of artificial cell culture micro element of the present invention mean flow rate with nutrient solution sample intake passage 14
Locate the curve map of change in flow.
Figure 13 is the growth of the inner cell of cell culture area 23 in different time sections of artificial cell culture micro element of the present invention
Condition diagram.
Figure 14 is that the 23 inner cell culture of cell culture area of artificial cell culture micro element of the present invention is used after 84 hours
Cell picture after DAPI dyeing.
Figure 15 is the growth curve chart of the inner cell of cell culture area 23 of artificial cell culture micro element of the present invention.
In figure:1 cell sample introduction layer;2 intermediate layers;3 bionical culture layers;4 nutrient solution sample holes;
5 cell sample holes;6 passages;7 alignment holes;8 cell sample outlet holes;9 waste liquid taps;
10 nutrient solution sample introduction through holes;11 culture pond intercommunicating pores;12 waste liquids discharge through hole;
13 nutrient solution liquid storage tanks;14 nutrient solution sample intake passages;15 bionical culture ponds;16 waste liquid sample output passages;
17 waste liquid liquid storage tanks;18 trapezoidal micro- dams;19 liquid storage areas;20 overflow areas;21 fence microtrabeculaes;
22 fence microtrabeculae gaps;23 cell culture areas;Microtrabeculae in 24;25 outer microtrabeculaes;
26 one-levels fork subchannel;27 2 fraction trouble passages;Microtrabeculae gap in 28.
Specific embodiment
Below in conjunction with accompanying drawing and technical scheme, specific embodiment of the invention is further illustrated.
Embodiment
As shown in figure 1, a kind of artificial cell culture micro element for building cell injuring model dynamic stability microenvironment,
The micro element is made up of three-decker, respectively cell sample introduction layer 1, intermediate layer 2 and bionical culture layer 3, cell sample introduction layer 1
Positioned at the superiors, intermediate layer 2 is located at centre, and bionical culture layer 3 is located at bottom.Nutrient solution sample introduction on cell sample introduction layer 1
Hole 4 is of the same size with the nutrient solution sample introduction through hole 10 on intermediate layer 2, a diameter of 0.5~1.5mm, the present embodiment
In be 1mm;Waste liquid discharge through hole 12 on waste liquid tap 9 and intermediate layer 2 on cell sample introduction layer 1 has identical chi
Very little, a diameter of 0.5~1.5mm is 1mm in the present embodiment;On intermediate layer 2 size shape of culture pond intercommunicating pore 11 with it is right
Neat hole 7, cell culture area 23 are identical, and shape can be square, regular hexagon, octagon or regualr decagon, polygonal
Inscribed circle diameter is 0.5~2mm, is the octagon of inscribed circle diameter 1mm in the present embodiment;The passage 6 of cell sample introduction layer upper 1
Symmetrical in pairs, the depth of passage is 30~100 microns, and the width of passage is 100~300 microns, passage 6 in the present embodiment
Depth and width be respectively 50 microns and 100 microns;Trapezoidal micro- dam 18 in the side of passage 6 being connected with cell sample outlet hole 8
The upper bottom and bottom of (as shown in Figure 3) are respectively 10~30 microns and 5~20 microns, and bottom and bottom are respectively 20 on the present embodiment
Micron and 10 microns, the height on trapezoidal micro- dam 18 is identical with the depth of passage, the present embodiment be 50 microns;In bionical culture layer 3
Nutrient solution liquid storage tank 13 a diameter of 1~3mm, in the present embodiment be 1.5mm, a diameter of 0.5~1.5mm of liquid storage area 19,
Identical with the size of nutrient solution sample holes 4, the outside diameter of the array of fence microtrabeculae 21 is 1.5mm~2.5mm, the week of fence microtrabeculae 21
It is 30~150 microns to width, is 90 microns, the height of fence microtrabeculae 21 and the depth of nutrient solution liquid storage tank 14 in the present embodiment
It is identical, it is 50~200 microns, it is 60 microns in the present embodiment;The width of nutrient solution sample intake passage 14 is 100~300 microns, this
Embodiment is 200 microns;One-level bifurcated passage 26 is identical with the depth of nutrient solution sample intake passage 14 with two grades of bifurcated passages 27, one-level
The width of bifurcated passage is calculated by its hydraulic diameter, and the cube sum of the hydraulic diameter of one-level fork subchannel 26 is equal to
The cube of the hydraulic diameter of nutrient solution sample intake passage 14, meets Murray laws, one-level bifurcated passage 26 in the present embodiment
Width is 69 microns, and the width of two fraction trouble passages 27 is identical with the width of one-level fork subchannel 26;Cell culture area 23
Be shaped as regular polygon, square, regular hexagon, octagon, regualr decagon, the inscribed circle diameter of regular polygon is 0.5
~2mm, for inscribed circle diameter is the octagon of 1mm in the present embodiment, depth is identical with nutrient solution sample intake passage 14;Cell is trained
The radial width of interior microtrabeculae 24 supported around area 23 is 50~200 microns, is 100 microns, the interior footpath of microtrabeculae gap 28 in the present embodiment
To the midline by the center of circle of cell culture area inscribed circle, circumferential width is 5~10 microns, is 6 microns, depth in the present embodiment
It is identical with cell culture area 23;Waste liquid sample output passage 16 is in inscribed circle of the nutrient solution sample intake passage 14 on cell culture area 23
It is symmetrical.Described bionical culture pond is desirably integrated into the grade fork microchannel network calculated according to Murray laws,
Realize the high-volume culture of cell.
Cell culture micro element measures the flowing of fluid in culture pond by method that trace particle tests the speed in embodiment
Speed.Specific operation, to deionized water is filled in micro element, is passed in and out cell using the good solid stopper of pre-production for first
Sample hole seals up, and then by silicone tube (internal diameter be 0.5mm~1.5mm, the present embodiment is 1mm) and pipe joint, (external diameter is
0.5mm~1.5mm, the present embodiment is 1mm) the nutrient solution sample holes of micro element are connected with the syringe above constant current syringe pump
Together, nutrient solution liquid outlet is passed through waste liquid cylinder by pipe joint and silicone tube.Micro element device is placed in inverted fluorescence microscope
Under, (concentration is 0.005~0.05g/ml to the well mixed microsphere suspension liquid of suction, and the present embodiment is 0.01g/ in syringe
Ml), the flow (1 μ L/min~15 μ L/min, the present embodiment is 5 μ L/min) of constant current syringe pump is set;Open constant current injection
Pump, adjusts microscopical brightness and focal length, image is focused at the half high position of the culture pond in bionical culture layer, to be seen
After the microballoon motion stabilization for arriving, recorded in intervals (20ms~500ms, the present embodiment is 50ms) by CCD camera
Picture;Using particles track resume module multiple continuous pictures in image processing software, obtain microballoon and transported in culture pond
Dynamic rail mark (as shown in Figure 10) and average movement velocity (as shown in figure 11).Change the flow of constant current syringe pump within the specific limits
(the present embodiment is 2 μ L/min, 3 μ L/min, 5 μ L/min, 7 μ L/min), represents micro element and is subject to the change for injecting pump discharge
External disturbance, measure it is different injection pump discharges under culture pond flow field change (as shown in figure 12).
Cell culture micro element, silicone tube and pipe joint autoclave sterilization 1 before for cell culture in embodiment
Hour, rat tail collagen protein is coated with after bacterium of having gone out (concentration is 0.01mg/ml~0.02mg/ml, the present embodiment 0.012mg/ml)
Or fibronectin (concentration is 1 μ g/ml~10 μ g/ml, the μ g/ml of the present embodiment 5), it is placed in incubator overnight after being coated with collagen,
It is adherent beneficial to subsequent cell.
Cell culture micro element is overnight passed through PBS and rinses out unnecessary albumen afterwards in coating collagen in embodiment, then inhales
Go out PBS, cell culture fluid is then filled in micro element, after treating that cell culture fluid is filled, will be advance by cell sample holes
(concentration is 5 × 10 to the Hela cell suspending liquids for making5~5 × 106Individual/ml, the present embodiment is 2 × 106Individual/ml) with 5 μ L/min
Injection pump discharge injection culture pond in, when cell sample outlet hole has liquid to emerge stop injection.A moment is stood, micro element is put
Situation is implanted into basis of microscopic observation cell, micro element is then put into CO2Incubator static incubation is adherent to most cells.
Cell culture micro element proceeds by dynamic cultivation after Hela cell attachments in embodiment, specifically by silica gel
Pipe and pipe joint link together micro element with syringe pump, are full of nutrient solution, setting note in syringe and silicone tube in advance
Penetrate pump discharge (2~20 μ L/min, the present embodiment be 5 μ L/min), open syringe pump, pushing syringe in micro element in a steady stream not
The fresh nutrient solution of disconnected input simultaneously discharges cell metabolism waste liquid.Cell culture micro element is during dynamic cultivation every 24
Hour is once observed take pictures (as shown in figure 13) the Hela cells in culture pond, after Hela cells are paved with whole culture pond
(as shown in figure 14) is dyeed with DAPI.The quantity of each time period inner cell is counted, cell growth curve is drawn (such as Figure 15 institutes
Show).
Claims (2)
1. a kind of multi-layer cellular culture micro element bionical based on vein eyed structure, it is characterised in that described multi-layer cellular
Culture micro element includes cell sample introduction layer (1), intermediate layer (2) and bionical culture layer (3), and cell sample introduction layer (1) is positioned at most
Upper strata, positioned at intermediate layer, bionical culture layer (3) is positioned at bottom for intermediate layer (2);
Described cell sample introduction layer (1) is provided with 4 through holes and 1 alignment hole, 4 through holes be respectively nutrient solution sample holes (4),
Cell sample holes (5), cell sample outlet hole (8) and waste liquid tap (9), alignment hole (7) is positioned at the center of cell sample introduction layer (1)
Place, nutrient solution sample holes (4) and waste liquid tap (9) are symmetrically positioned in alignment hole (7) left and right end, cell sample holes (5) and cell
Sample outlet hole (8) is symmetrically positioned in alignment hole (7) upper and lower side;Between cell sample holes (5) and cell sample outlet hole (8) by passage (6) even
Logical, passage (6) is arranged symmetrically in pairs;Passage (6) side being connected with cell sample outlet hole (8) is transversely provided with trapezoidal micro- dam (18), uses
Culture pond is rushed out in stop cell;
Described intermediate layer (2) is provided with 3 through holes, is from left to right followed successively by nutrient solution sample introduction through hole (10), culture pond
Intercommunicating pore (11) and waste liquid discharge the nutrient solution sample holes of through hole (12), nutrient solution sample introduction through hole (10) and cell sample introduction layer (1)
(4) communicate, waste liquid discharge through hole (12) with cell sample introduction layer (1) waste liquid tap (9) communicate, culture pond intercommunicating pore (11) and
The alignment hole (7) of cell sample introduction layer (1) is relative, and shape size is identical;
Described bionical culture layer (3) is provided with nutrient solution liquid storage tank (13), bionical culture pond (15) and waste liquid liquid storage tank (17),
Nutrient solution enters nutrient solution liquid storage tank (13), nutrient solution liquid storage from nutrient solution sample holes (4) by nutrient solution sample introduction through hole (10)
It is connected by nutrient solution sample intake passage (14) between pond (13) and bionical culture pond (15), bionical culture pond (15) and waste liquid liquid storage tank
(17) it is connected by waste liquid sample output passage (16) between;
Described nutrient solution liquid storage tank (13) is divided into liquid storage area (19) and overflow area (20), and liquid storage area (19) is positioned at nutrient solution liquid storage
The penetralia in pond (13), liquid storage area (19) is surrounded by multiple fence microtrabeculaes (21), and the inscribed circle diameter of liquid storage area (19) is not less than
The inscribed circle diameter of nutrient solution sample introduction through hole (10), nutrient solution enters overflow area (20) by fence microtrabeculae gap (22), overflow
Area (20) communicates with nutrient solution sample intake passage (14);
Described bionical culture pond (15) is provided with two-layer microtrabeculae, is interior microtrabeculae layer and outer microtrabeculae layer, and interior microtrabeculae layer is arranged by multiple
The cloth polygonized structure that closely interior microtrabeculae (24) is surrounded, the polygonized structure inside surrounded by interior microtrabeculae (24) is cell culture
Area (23), nutrient solution enters cell culture area (23) by the interior microtrabeculae gap (28) between interior microtrabeculae (24);Outer microtrabeculae layer is located at
Interior microtrabeculae layer outside, the quantity of the outer microtrabeculae (25) arranged in outer microtrabeculae layer is identical with polygonal side number, by bionical culture pond
(15) internal passage is divided into one-level fork subchannel (26) and two fraction troubles passage (27);One-level diverge subchannel (26) be
The passage formed between passage, outer microtrabeculae layer China and foreign countries's microtrabeculae (25) that bionical culture pond (15) is formed with outer microtrabeculae interlayer, a fraction
The hydraulic diameter of trouble passage (26) meets Murray laws, i.e. the cube sum of one-level fork subchannel (26) hydraulic diameter
Equal to the cube of nutrient solution sample intake passage (14) water conservancy diameter;Two fraction troubles passage (27) are outer microtrabeculae layer and interior microtrabeculae layer
Between the passage that is formed, two fraction troubles passage (27) are afterbody subchannel, the simulation most thin vein of blade, are diverged with one-level
Subchannel (26) is with identical hydraulic diameter.
2. multi-layer cellular culture micro element according to claim 1, it is characterised in that described cell sample introduction layer (1), in
Between transition zone (2) and bionical culture layer (3) material be thermosetting polymer dimethyl silicone polymer, the poly- first of thermoplastic polymer
Base methyl acrylate, polystyrene or makrolon.
Priority Applications (1)
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CN111635545A (en) * | 2020-05-28 | 2020-09-08 | 大连理工大学 | Bionic orientation blade structure material for capturing tumor cells and preparation method thereof |
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