CN201581070U - Chip for cell pairing and fusion - Google Patents

Chip for cell pairing and fusion Download PDF

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Publication number
CN201581070U
CN201581070U CN 200920218980 CN200920218980U CN201581070U CN 201581070 U CN201581070 U CN 201581070U CN 200920218980 CN200920218980 CN 200920218980 CN 200920218980 U CN200920218980 U CN 200920218980U CN 201581070 U CN201581070 U CN 201581070U
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cell
lune
opening
chip
fusion
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Expired - Fee Related
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CN 200920218980
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Chinese (zh)
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陈慧敏
徐念沁
顾传虎
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JIANGYIN STEMEASY BIOLOGICAL TECHNOLOGY CO LTD
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JIANGYIN STEMEASY BIOLOGICAL TECHNOLOGY CO LTD
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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
    • C12M47/00—Means for after-treatment of the produced biomass or of the fermentation or metabolic products, e.g. storage of biomass
    • C12M47/06—Hydrolysis; Cell lysis; Extraction of intracellular or cell wall material

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Abstract

The utility model relates to a chip for cell pairing and fusion, belonging to the technical field of biomedicine. The chip for cell pairing and fusion comprises a chip body. A hollow closed cell fusion chamber is arranged in the chip. A cell well is arranged on the bottom plate of the cell fusion chamber and is formed by two symmetrical half-moon dikes. Each half-moon dike is provided with a short end and a long end. The protruded arc surfaces of the two half-moon dikes face to each other. The short ends of the two half-moon dikes form a first opening and the long ends of the two half-moon dikes form a second opening. The bottom part of each half-moon dike is fixed on the bottom plate and a certain clearance is kept between the top part of each half-moon dike and the top plate of the cell fusion chamber. A first sample adding hole corresponding to the direction of the first opening of the cell well and a second sample adding hole corresponding to the direction of the second opening of the cell well are arranged in the direction of two opposite corners of the cell fusion chamber. The utility model has the advantages that the structure is simple and ingenious, the cell pairing rate and the cell fusion rate are greatly improved, the chemical toxic effect of inducers on cells is reduced, and the operation is simple and convenient.

Description

Cell pairing and fusion chip
Technical field
The utility model belongs to field of biomedicine technology, relates to a kind of device that can realize that the cell pairing is merged, specifically a kind of cell pairing and fusion chip.
Background technology
Cytogamy (cell fusion) is meant under field conditions (factors) or makes two or more cells merge the process that forms a cell with manual method (biological, physics, chemistry).Its value that in Application Areass such as genetics, developmental biology, animals and plants distant hybridization breeding, immune medical science and medicine, food, agricultural, all has a wide range of applications.Some advanced technology in the modern biomedical, as in the production of monoclonal antibody, mammiferous clone, microbial strains seed selection, drafting gene mapping and anti-tumor vaccine are produced, cytogamy is wherein the most key step.
Although in the many decades in the past, cell-fusion techniques has been obtained tremendous development, has obtained remarkable achievement, still has many problems, has restricted the widespread use of cell-fusion techniques and further development greatly.The method that inducing cell merges mainly contains three kinds:
The one, biological method, its inductor adopt usually through the virus of inactivation treatment such as Sendai virus (Sendai virus) etc., and this method generally is that the zooblast fusion is peculiar.
The 2nd, physical method, it adopts, and electricity swashs, the induced with laser cytogamy; This method is used in current domestic a few experiments chamber, electricity induces integration technology (cell electrofusion) to be invented by U.S. scientist Zimmerman, and he has carried out systematic research in 20th century eight, the nineties pair cell electricity integration technology, started the new situation of cell electricity integration technology, made cell-fusion techniques become bionic taproot technology.But equally also there is following deficiency in it: the specificity pairing rate of (1), cell is low, thereby causes cell confluency generally can only reach about 10%; (2), owing to being in conduction, electrode merges in the liquid, therefore between electrode, set up enough strength of electric field, will export and be at least several hectovolts even thousands of volt, just require power supply that high output rating is arranged, so just cause cell electricity fusion device price high, and have the hidden danger of electrical safety aspect, thereby hindered this broad application.
The 3rd, chemical process, its inductor adopts polyoxyethylene glycol (Polyethyleneglycol, PEG), it is very low to utilize the PEG method to carry out the cost of cytogamy, use also the most extensive, but this integration technology also exists and much is difficult to the shortcoming that overcomes: the pairing of (1), specificity is poor, and fusion efficiencies is low, and the specificity fusion rate of different cells generally is not higher than 1%; (2), the cell fusion process poor controllability, repeatable poor; (3) because the specificity pairing is poor, and the usage quantity of inductor PEG is also bigger, and the inductor pair cell has bigger chemical toxic action, and consumption is big more, and toxic action is strong more, is unfavorable for the existence and the function of fused cell.
Summary of the invention
The purpose of this utility model is to overcome the deficiencies in the prior art, a kind of cell pairing and fusion chip of simple and ingenious structure is provided, this chip can greatly improve cell pairing rate and fusion rate, reduces the chemical toxic action of inductor pair cell, and simple to operation.
The technical scheme that provides according to the utility model, described cell pairing and fusion chip comprises the chip body, the chip body is provided with the airtight cytogamy chamber of hollow, the base plate of cytogamy chamber is provided with the cell trap, the cell trap is made of two symmetric lune burrocks, the lune burrock has short end and long end, the convex globoidal of two lune burrocks is relative, their short end forms first opening, first opening only can hold individual cells, their long end forms second opening, and second opening only can hold two cells; The bottom of lune burrock is fixed on the base plate, and the top of lune burrock and cytogamy chamber top board leave a determining deviation; The cytogamy chamber the angular direction is offered second well of second opening direction of first well of first opening direction of corresponding cell trap and corresponding cell trap.
The short end length L of described lune burrock 1Be 5～50 μ m, long end length L 2Be 10～100 μ m, thickness is 5～50 μ m.
The spacing d of two lune burrocks in the described cell trap is smaller or equal to 100 μ m; Preferably, the spacing d of two lune burrocks is 5～30 μ m.
Described lune burrock paste fixing or direct engraving on the base plate of cytogamy chamber.
As further improvement of the utility model, outwards extended funnel-shaped opening on described first well and second well respectively, so that the filling cell suspension; Preferably, described funnel-shaped opening is equipped with corresponding lid.
As further improvement of the utility model, the inwall of described cytogamy chamber and lune burrock are through polished finish, in case pair cell causes physical abuse.
As further improvement of the utility model, the short end of described lune burrock and the termination portion of long end are made as circular arc, in case pair cell causes physical abuse.
The utility model simple and ingenious structure, compared with prior art, advantage is:
(1), special cell well structure makes and holds a pair of cell to be merged in each cell trap, greatly improve the specificity pairing rate and the cell confluency of cell.
(2), because the pairing rate improves, the usage quantity of inductor PEG also significantly reduces, thereby has reduced the chemical toxic action of inductor pair cell.
(3), the lune burrock in the cell trap adopts and to paste mode and fix, by adjusting the size that two angles between the lune burrock can change first opening, second opening, the size of cell trap and ranks spacing are also adjustable flexibly, to adapt to different cells, reduce manufacturing cost.
(4), well outwards extended funnel-shaped opening, the filling cell suspension is more simple and convenient, reduces operation easier greatly, reduces use cost.
(5), the inwall of cytogamy chamber and lune burrock be through polished finish, the short end of lune burrock and the termination portion of length are made as circular arc, thereby can not cause physical abuse by pair cell.
Description of drawings
Fig. 1 is a three-dimensional arrangement view of the present utility model.
Fig. 2 is a plan view of the present utility model.
Fig. 3 is the structural representation of a cell trap in the utility model.
Description of reference numerals: 1-chip body, 2-first well, 3-base plate, 4-top board, 5-second well, 6-funnel-shaped opening, 7-lid, 8-lune burrock, 9-cell trap, 10-first opening, 11-second opening.
Embodiment
The utility model is described in further detail below in conjunction with concrete drawings and Examples.
As Fig. 1～shown in Figure 3, the utility model comprises chip body 1, and chip body 1 is provided with the airtight cytogamy chamber of hollow, and the base plate 3 of cytogamy chamber is provided with the cell trap 9 of some amount, forms cell trap 9 arrays; Cell trap 9 is made of two symmetric lune burrocks 8, lune burrock 8 has short end and long end, the convex globoidal of two lune burrocks 8 is relative, their short end forms first opening 10, first opening 10 only can hold individual cells, their long end forms second opening, 11, the second openings 11 and only can hold two cells; The bottom of lune burrock 8 is fixed on the base plate 3, and the top of lune burrock 8 and cytogamy chamber top board 4 leave a determining deviation; The cytogamy chamber the angular direction is offered second well 5 of second opening, 11 directions of first well 2 of first opening, 10 directions of corresponding cell trap 9 and corresponding cell trap 9.
As shown in Figure 1 and Figure 2, the size of considering cell generally all is a micron order, correspondingly, the size that is opened in first well 2 on the chip body 1 and second well 5 is also very little, the difficulty that directly drips cell suspension in first well 2 and second well 5 is bigger, so the utility model has outwards extended funnel-shaped opening 6 respectively on first well 2 and second well 5, funnel-shaped opening 6 can be suitable with liquid feeding instrument such as liquid-transfering gun, and is more convenient when making the filling cell suspension; Simultaneously, the funnel-shaped opening 6 on first well 2 and second well 5 can set a lid 7, and in time spent not cell pairing and fusion chip free time, lid 7 has improved security.
As Fig. 2, shown in Figure 3, the size of described lune burrock 8 can be chosen according to the cell of required fusion, in general, and short end length L 1Be 5～50 μ m, long end length L 2Be 10～100 μ m, thickness is 5～50 μ m.The spacing d of two lune burrocks 8 in the described cell trap 9 generally should be smaller or equal to 100 μ m, and preferably, the spacing d of two lune burrocks 8 is 5～30 μ m.
Described lune burrock 8 is fixed on the base plate 3 of cytogamy chamber, this fixing traditional direct engraving mode that both can adopt, also can adopt and paste fixed form, but because the manufacturing cost of direct engraving is very high, so preferred employing is pasted fixing in the utility model, manufacturing cost is reduced greatly, and make the contour structures of cell trap 9 more flexible and changeable, adaptability is stronger.
The inwall and the lune burrock 8 of described cytogamy chamber all pass through polished finish, and the short end of lune burrock 8 and the termination portion of long end are made as circular arc, in case pair cell causes physical abuse.
Principle of the present utility model and using method are as follows: (with the pairing of Chinese hamster ovary celI and 293T cell and be fused to example)
(1) chip is stood on the desktop, make a diagonal lines that offers first, second well 2,5 perpendicular to desktop, guarantee that first opening 10 of the cell trap 9 in the chip body 1 is directed upwardly, first well 2, second well 5 stay open state; It is indoor with liquid-transfering gun the Chinese hamster ovary celI suspension to be added dropwise to cytogamy from first well 2 then, slightly rock simultaneously chip, because action of gravity and concussion effect, cell suspension enters respectively in first opening 10 of each cell trap 9, because first opening 10 only can hold and possess individual cells, other cell just enters next cell trap 9, by that analogy, possesses a cell respectively in first opening 10 of each the cell trap 9 in the chip body 1;
(2) with chip Rotate 180 ° and standing on the desktop rapidly in himself plane, first well 2 down at this moment, second well 5 up, because action of gravity, be retained in single Chinese hamster ovary celI landings in first opening 10 originally and fall in the second bigger opening 11 of another cell trap 9 of its below and be retained;
(3) it is indoor the 293T cell suspension to be added dropwise to cytogamy with liquid-transfering gun from second well 5, similarly slightly rock chip, the 293T cell suspension is evenly entered in the cell trap 9 in the chip body 1, because second opening 11 of cell trap 9 only can hold two cells, after holding a Chinese hamster ovary celI, can only hold a 293T cell again, like this, two cells to be merged just are maintained in the same cell trap 9;
(4) in cytogamy is indoor, drip inductor PEG by second well 5 again, slightly rock chip simultaneously, to induce Chinese hamster ovary celI and 293T cytogamy; After dripping PEG, leave standstill about 90 seconds, drip cell culture medium then, what just begun to drip is slower, can add fast behind the 1min, and then leave standstill about 10min, to finish cytogamy;
(5) with chip once more 180 ° of himself plane internal rotation, add a large amount of cell culture mediums from first well 2 again, will merge good cell wash-outs, recovery in the cell trap 9.
Cell pairing and fusion chip of the present utility model is not limited only to the mixing operation of above-mentioned two kinds of cells, and it goes for any two kinds of cells.The relative position of two lune burrocks 8 in cell trap 9 can design, make according to the size and the experimental requirements of the cell that will merge, as changes relative angle, distance of two lune burrocks 8 etc.

Claims (9)

1. cell pairing and fusion chip, comprise chip body (1), it is characterized in that, chip body (1) is provided with the airtight cytogamy chamber of hollow, the base plate of cytogamy chamber (3) is provided with cell trap (9), cell trap (9) is made of two symmetric lune burrocks (8), lune burrock (8) has short end and long end, the convex globoidal of two lune burrocks (8) is relative, their short end forms first opening (10), first opening (10) only can hold individual cells, and their long end forms second opening (11), and second opening (11) only can hold two cells; The bottom of lune burrock (8) is fixed on the base plate (3), and the top of lune burrock (8) and cytogamy chamber top board (4) leave a determining deviation; The cytogamy chamber the angular direction is offered second well (5) of second opening (11) direction of first well (2) of first opening (10) direction of corresponding cell trap (9) and corresponding cell trap (9).
2. cell pairing and fusion chip as claimed in claim 1, its feature also are, the short end length (L of described lune burrock (8) 1) be 5～50 μ m, long end length (L 2) be 10～100 μ m, thickness is 5～50 μ m.
3. cell pairing and fusion chip as claimed in claim 1, its feature are that also the spacing (d) of two the lune burrocks (8) in the described cell trap (9) is smaller or equal to 100 μ m.
4. cell pairing and fusion chip as claimed in claim 1, its feature are that also the spacing (d) of two the lune burrocks (8) in the described cell trap (9) is 5～30 μ m.
5. cell pairing and fusion chip as claimed in claim 1, its feature also are, have outwards extended funnel-shaped opening (6) respectively on described first well (2) and second well (5).
6. cell pairing and fusion chip as claimed in claim 1, its feature are that also described lune burrock (8) is pasted on the base plate (3) that is fixed on the cytogamy chamber.
7. cell pairing and fusion chip as claimed in claim 1, its feature are that also described lune burrock (8) direct engraving is on the base plate (3) of cytogamy chamber.
8. cell pairing and fusion chip as claimed in claim 1, its feature are that also the short end of described lune burrock (8) and the termination portion of long end are made as circular arc.
9. cell pairing and fusion chip as claimed in claim 5, its feature are that also described funnel-shaped opening (6) is equipped with corresponding lid (7).
CN 200920218980 2009-12-11 2009-12-11 Chip for cell pairing and fusion Expired - Fee Related CN201581070U (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102174391A (en) * 2011-03-10 2011-09-07 北京航空航天大学 Cell electrofusion chip
CN102504997A (en) * 2011-11-01 2012-06-20 彭兴跃 Chip for cell observation and experiments

Cited By (4)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102174391A (en) * 2011-03-10 2011-09-07 北京航空航天大学 Cell electrofusion chip
CN102174391B (en) * 2011-03-10 2014-05-07 北京航空航天大学 Cell electrofusion chip
CN102504997A (en) * 2011-11-01 2012-06-20 彭兴跃 Chip for cell observation and experiments
CN102504997B (en) * 2011-11-01 2016-08-17 彭兴跃 A kind of cell observation experiment chip

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Granted publication date: 20100915

Termination date: 20121211