CN102061252A - Aseptic simple liquor relief device of cell culture liquid - Google Patents

Aseptic simple liquor relief device of cell culture liquid Download PDF

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Publication number
CN102061252A
CN102061252A CN2010105638453A CN201010563845A CN102061252A CN 102061252 A CN102061252 A CN 102061252A CN 2010105638453 A CN2010105638453 A CN 2010105638453A CN 201010563845 A CN201010563845 A CN 201010563845A CN 102061252 A CN102061252 A CN 102061252A
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aseptic
cell culture
liquid
rifle head
dash
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王旭
李文哲
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Dalian University
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Dalian University
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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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Abstract

The invention relates to an aseptic simple liquor relief device of a cell culture liquid, which is characterized in that a rubber hose is connected with a spearhead connector first, then respectively connected with upper pipe orifices of two intubation tubes inserted into a waste liquid collecting bottle from a plug, and then connected with a gas divider and a gas pump. The front end of the spearhead connector is contracted and thinned so as to insert the tenuous spearhead. The gas divider is a three-way pipe in a y-shaped structure. Closable elastic membranes with V-shaped sections and upward closed angles are respectively in the middle lower portion of the long dash and in the middle portion of short dash of the y-shaped structure. The included angle between the long dash and the short dash of the y-shaped structure is ordinarily 30-60 degrees, preferably 45 degrees. The aseptic simple liquor relief device of the cell culture liquid makes the liquid absorption process more accurate and efficient and is simpler and faster for operation; and the liquid is absorbed cleanly and thoroughly, thereby eliminating the corrupt practices of artificial pollution and the like.

Description

The aseptic simple and easy liquid-transfering device of cell culture fluid
Technical field
The aseptic transfer facility of the present invention cell culture fluid and other liquid in the iuntercellular super clean bench.
Background technology
In cell cultivation process, often wash operations such as cell, peptic cell and replacing nutrient solution, with the normal growth that guarantees cell and the cultivation of going down to posterity, very strict to its aseptic requirement.Present method is when changing liquid at every turn, need to use the glass pipette of Amoxcillin with cell washing lotion and old nutrient solution sucking-off, change aseptic suction pipe again and draw new nutrient solution to culturing bottle, and the liquid of sucking-off will temporarily be put into super clean bench waste liquid cylinder, again it is brought out after experiment finishes, put back to super clean bench after toppling over processing.This traditional working method is loaded down with trivial details time-consuming, and because the restriction of its suction nozzle of glass pipette can cause liquid assimilating unclean, topples over the waste liquid cylinder repeatedly and also be easy to generate artificial drawbacks such as pollution.
Summary of the invention
Technical problem to be solved by this invention provides the aseptic liquid-transfering device of a kind of cell culture fluid and other liquid, and this device can be simplified the operating process of absorption enchylema, and can make this operating process more accurate, convenient, efficient.
For solving the problems of the technologies described above, the following technical scheme of proposition of the present invention: the aseptic simple and easy liquid-transfering device of a kind of cell culture fluid, its feature is: connect the rifle head connector earlier, connect the upper orifice that inserts two intubate of waste collection bottle from bottle stopper respectively again with sebific duct, connect gas diverter and air pump afterwards.The front end of its rifle head connector shrinks and attenuates, so that insert elongated rifle head.Its gas diverter is the Y-tube of y font structure, the middle and lower part of y font dash and short draw the middle part be respectively equipped with wedge angle consistent towards upper section be V-arrangement can closed elastica lobe.This gas diverter is the Y-tube of y font structure, and the angle between y font dash and short the drawing is generally 30～60 °, is preferably 45 °.
Adopt the aseptic simple and easy liquid-transfering device of cell culture fluid of technique scheme, make the liquid assimilating process more accurately, simple and fast during efficient, operation, the absorption of liquid is totally thorough, has eliminated drawbacks such as artifact pollution.
Description of drawings
Now in conjunction with the accompanying drawings the present invention is elaborated:
Fig. 1 is the aseptic simple and easy liquid-transfering device system schematic of cell culture fluid of the present invention
Fig. 2 is the enlarged view of preface 2 parts among Fig. 1
Fig. 3 is the enlarged view of preface 7 parts among Fig. 1
Embodiment
As shown in Figure 1: the aseptic simple and easy liquid-transfering device of nutrient solution of the present invention, it connects rifle head connector 2 earlier, connects the suitable for reading of the pipe 4 of filling in waste collection bottle 6 from bottle stopper 3 and pipe 5 respectively again with sebific duct 1, connects gas diverter 7 and air pump 8 then.
Sebific duct 1 is a medical latex tube, and its length is decided by specific requirement.Its rifle head connector 2 front ends shrink and attenuate, so that insert elongated rifle head.Rifle head connector 2 can be made of common metal tube.Its front end point mouth place can connect the elongated rifle head of sterilization then in the place's calcination of spirit lamp flame, is used for putting in the Tissue Culture Flask imbitition.It is tiny and sharp-pointed that the suction nozzle of rifle head is compared traditional glass pipette, therefore eliminated the sordid phenomenon of using in the past of method liquid assimilating.Remove rifle head connector and part sebific duct and can be placed on super clean bench, all the other each ones all can place outside the super clean bench.Waste collection bottle 6 is the triangular flask of 5L.Its gas diverter 7 is the Y-tube of y font structure, and the material of tube wall can be selected metal, pottery, glass etc. for use, generally selects Glass tubing.At the middle and lower part of y font dash and the short middle part of drawing, be respectively equipped with the wedge angle unanimity towards upper section be V-arrangement can closed elastica lobe 9 and 10.They are equivalent to the reverse checkvalve in the tubing system.This gas diverter is the Y-tube of y font structure, and the angle between y font dash and short the drawing is generally 30～60 °, is preferably 45 °.
Pipe 4 upper ends of inserting 3 li of bottle closure of rubber are connected with waste liquid derivation sebific duct 1, and the upper end of another pipe 5 is connected by sebific duct 1 with air pump 8.Air pump 8 connects with gas diverter 7 usefulness sebific ducts 1, just can operate.After stepping on treadle air pump, leading to extraneous V-type lobe 10 can open, and gas is discharged; Pin leaves treadle air pump, and leading to extraneous V-type lobe 10 can be closed.Because at this moment form negative pressure in the total system, this causes the V-type lobe 9 in another branch to be opened, liquid is sucked into the rifle head, through rifle head connector 2---sebific duct 1---intubate 4 enters in the waste collection bottle 6.Air pump 8 is that difunctional foot-operated aerator pump is taken out in common common filling, and compares simple and easy, the consumed power, pollution-free not of power air pump operation.
For further setting forth the present invention, enumerate three specific embodiments below and introduce using method of the present invention.
Embodiment 1 carries out suspension cell and goes down to posterity when cultivating:
The first step: the sharp mouth of calcination rifle head connector, treat that temperature lowers to plug aseptic rifle head, unsettled placement.
Second step: the cell in the Tissue Culture Flask is transferred to the 15ml centrifuge tube with suction pipe, the centrifugal cell precipitation that makes.Pin is stepped on air pump, unclamps, and the rifle head puts in centrifuge tube liquid, and the waste collection bottle is advanced in the liquid pumpback.
The 3rd step: draw a certain amount of PBS damping fluid to centrifuge tube, pressure-vaccum cell several times, centrifugal after, the rifle head connector is changed and to be extracted liquid by above operation behind the rifle head and arrive the waste collection bottle.
The 4th step: draw fresh medium to centrifuge tube, blow and beat mixing gently, on average assign to two culturing bottles then.
Table 1 adopts traditional method and the present invention to carry out suspension cell and goes down to posterity culture effect relatively
Figure BDA0000034772000000031
Embodiment 2 carries out going down to posterity when cultivating of attached cell:
The first step: the sharp mouth of calcination rifle head connector, treat that temperature lowers to plug aseptic rifle head, unsettled placement.
Second step: pin is stepped on air pump, unclamps, and the rifle head puts in the Tissue Culture Flask liquid, and the waste collection bottle is advanced in the liquid pumpback.
The 3rd step: draw a certain amount of PBS damping fluid to culturing bottle, the purge several times extract liquid to the waste collection bottle by above operation behind the rifle head connector replacing rifle head gently.Repeat this step, influence of serum reduces to minimum when guaranteeing trysinization.
The 4th step: draw a small amount of trysinization liquid to culturing bottle, digest and draw the effect of fresh medium after 1 minute, in case the injury cell to culturing bottle termination pancreatin.Piping and druming moves to the 15ml centrifuge tube gently, and is centrifugal, and the rifle head connector extracts liquid to the waste collection bottle by above operation after changing the rifle head.
The 5th step: draw fresh medium to centrifuge tube, blow and beat mixing gently, on average assign to two culturing bottles then.
Table 2 adopts traditional method and the present invention to carry out attached cell and goes down to posterity culture effect relatively
Figure BDA0000034772000000032
When embodiment 3 carries out lysis,
The first step: the sharp mouth of calcination rifle head connector, treat that temperature lowers to plug aseptic rifle head, unsettled placement.
Second step: pin is stepped on air pump, unclamps, and the rifle head puts in the Tissue Culture Flask liquid, and the waste collection bottle is advanced in the liquid pumpback.
The 3rd step: draw a certain amount of PBS damping fluid to culturing bottle, the purge several times extract liquid to the waste collection bottle by above operation behind the rifle head connector replacing rifle head gently.Repeat this step, influence of serum reduces to minimum when guaranteeing trysinization.
The 4th step: draw trysinization liquid to culturing bottle, digest and draw the effect of fresh medium after 1 minute, in case the injury cell to culturing bottle termination pancreatin.Piping and druming moves to the 15ml centrifuge tube gently, and is centrifugal, and the rifle head connector extracts liquid to the waste collection bottle by above operation after changing the rifle head.
The 5th step: wash cell twice with the 3rd step PBS, influence of serum reduces to minimum when guaranteeing lysis.
The 6th step: the cell pyrolysis liquid lysing cell, carry out subsequent experimental.
Table 3 adopts traditional method and the present invention to carry out the lysis effect relatively
Figure BDA0000034772000000041

Claims (5)

1. aseptic simple and easy liquid-transfering device of cell culture fluid, it is characterized in that: connect rifle head connector (2) earlier, connect respectively from the intubate (4) of bottle stopper (3) insertion waste collection bottle (6) and the upper orifice of (5) again with sebific duct (1), connect gas diverter (7) and air pump (8) afterwards.
2. the aseptic simple and easy liquid-transfering device of cell culture fluid according to claim 1 is characterized in that: the contraction of described rifle head connector (2) front end attenuates and inserts elongated rifle head.
3. the aseptic simple and easy liquid-transfering device of cell culture fluid according to claim 1, it is characterized in that: described gas diverter (7) is the Y-tube of y font structure, at the middle and lower part of y font dash and the short middle part of drawing, be respectively equipped with the wedge angle unanimity towards upper section be V-arrangement can closed elastica lobe (9) and (10).
4. the aseptic simple and easy liquid-transfering device of cell culture fluid according to claim 3 is characterized in that: described gas diverter (7) is the Y-tube of y font structure, and the angle between described y font dash and short the drawing is 30～60 °.
5. the aseptic simple and easy liquid-transfering device of cell culture fluid according to claim 4 is characterized in that: described gas diverter (7) is the Y-tube of y font structure, and the angle between described y font dash and short the drawing is preferably 45 °.
CN2010105638453A 2010-11-29 2010-11-29 Aseptic simple liquor relief device of cell culture liquid Pending CN102061252A (en)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102823430A (en) * 2012-09-17 2012-12-19 平慧芳 Device for filling culture solution into test tube
CN103563859A (en) * 2013-11-18 2014-02-12 华中农业大学 Device and method for replacing nutrient solution for indoor cultivation of daphnia magna
CN105462833A (en) * 2015-12-31 2016-04-06 苏州壹达生物科技有限公司 Method for controlling even and stable solution flow speed by compressing air through peristaltic pump
CN108339814A (en) * 2018-04-25 2018-07-31 洛阳师范学院 A kind of device quickly cleaning solid medium in conical flask
JP2019205399A (en) * 2018-05-30 2019-12-05 学校法人国士舘 Cell-treating method, device and system

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN86210246U (en) * 1986-12-15 1988-03-16 楼许柏 Multipurpose liquid aspirator
CN2201065Y (en) * 1994-08-31 1995-06-21 孟凡允 Active valve foetal head sucker
CN2352721Y (en) * 1998-12-28 1999-12-08 顾忠祥 Air sack type pipet
CN201316454Y (en) * 2008-11-07 2009-09-30 王凤兰 Quick pressurizing infusion device for high infusion volume
CN201410389Y (en) * 2009-06-04 2010-02-24 杨志昌 Transfer burette

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN86210246U (en) * 1986-12-15 1988-03-16 楼许柏 Multipurpose liquid aspirator
CN2201065Y (en) * 1994-08-31 1995-06-21 孟凡允 Active valve foetal head sucker
CN2352721Y (en) * 1998-12-28 1999-12-08 顾忠祥 Air sack type pipet
CN201316454Y (en) * 2008-11-07 2009-09-30 王凤兰 Quick pressurizing infusion device for high infusion volume
CN201410389Y (en) * 2009-06-04 2010-02-24 杨志昌 Transfer burette

Cited By (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102823430A (en) * 2012-09-17 2012-12-19 平慧芳 Device for filling culture solution into test tube
CN102823430B (en) * 2012-09-17 2013-10-30 平慧芳 Device for filling culture solution into test tube
CN103563859A (en) * 2013-11-18 2014-02-12 华中农业大学 Device and method for replacing nutrient solution for indoor cultivation of daphnia magna
CN105462833A (en) * 2015-12-31 2016-04-06 苏州壹达生物科技有限公司 Method for controlling even and stable solution flow speed by compressing air through peristaltic pump
CN105462833B (en) * 2015-12-31 2019-01-04 苏州壹达生物科技有限公司 A method of it is uniform and stable using peristaltic pump compressed air control solution flow velocity
CN108339814A (en) * 2018-04-25 2018-07-31 洛阳师范学院 A kind of device quickly cleaning solid medium in conical flask
JP2019205399A (en) * 2018-05-30 2019-12-05 学校法人国士舘 Cell-treating method, device and system
JP7202080B2 (en) 2018-05-30 2023-01-11 テルモ株式会社 Cell processing method, device and system

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Application publication date: 20110518