CN105112289A - Cell separation device and dispenser applied to same - Google Patents
Cell separation device and dispenser applied to same Download PDFInfo
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- CN105112289A CN105112289A CN201510560187.5A CN201510560187A CN105112289A CN 105112289 A CN105112289 A CN 105112289A CN 201510560187 A CN201510560187 A CN 201510560187A CN 105112289 A CN105112289 A CN 105112289A
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- cell separation
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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
- C12M47/04—Cell isolation or sorting
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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
- C12M37/00—Means for sterilizing, maintaining sterile conditions or avoiding chemical or biological contamination
- C12M37/02—Filters
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- Separation Using Semi-Permeable Membranes (AREA)
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Abstract
The invention provides a cell separation device and a dispenser applied to the same. The cell separation device comprises a vertical spindle, a horizontal centrifugal arm and cell separation collectors, wherein the centrifugal arm is connected to the spindle and rotates around the spindle, and the cell separation collectors are fixedly connected to at least one end of the centrifugal arm. The dispenser is arranged above the spindle and comprises a liquid inlet pipeline, a liquid inlet cavity, a liquid outlet pipeline and a liquid outlet cavity, the liquid inlet cavity is communicated with the liquid inlet pipeline, and the liquid outlet cavity is communicated with the liquid outlet pipeline. Infusion openings are formed in the liquid inlet cavity and the liquid outlet cavity respectively. Each cell separation collector comprises a liquid inlet communicated with the infusion opening of the liquid inlet cavity, a liquid outlet communicated with the infusion opening of the liquid outlet cavity, a collection cavity, a waste liquid cavity and a filter membrane, wherein the collection cavity and the waste liquid cavity are communicated to the liquid inlet and the liquid outlet respectively, and the filter membrane is arranged between the collection cavity and the waste liquid cavity. The diameters of filter holes of the filter membranes are smaller than the diameters of to-be-separated cells. The cell separation device is simple in structure, convenient to operate and capable of separating cells quickly and accurately, separation is performed in a totally-closed system, and damage and pollution to the cells are not caused easily.
Description
Technical field
The invention belongs to medical instruments field, be particularly a kind ofly separated for dynamic centrifugal and collect the cell separation apparatus of useful cell and the skimmer for this cell separation apparatus.
Background technology
Cell therapy refers to the characteristic utilizing some to have the cell of specific function, after employing biological engineering method is obtained and/or is processed by amplification in vitro, specific culture etc., make these cells have enhancing immunity, kill the therapeutic efficiency such as pathogenic agent and tumour cell, promotion tissue and organ regeneration and physical recovery, thus reach the object of disease therapy.Cell therapy is T&B, is widely used in tumour, hepatic diseases at present, goes the every field such as scar, and have vast potential for future development.
The most basic step of cell therapy is exactly first will isolate target cell.In prior art, roughly there are two kinds of methods to realize the object of isolated cell.The first directly aspirates mixing liquid with micro-pore-film filtration pumping equipment, and small-particle and certain liquid are drained by filter membrane under suction function, and particle big or middle is trapped together with major diameter cell, thus realizes major diameter cell with short grained and be separated; The second whizzer is directly separated mixing liquid, thus realizes being separated of major diameter cell and large and small particle; The weak point of first method is, because the pore size filter of microporous membrane is little in during negative pressure-pumping, if strengthened it; film strength is inadequate, easily breaks, therefore; often because of the fenestra of the too many and blocking microporous film of the small-particle in mixing liquid; easily cause and be separated not exclusively or be separated unsuccessfully, and macrobead also needs otherwise processed, the weak point of second method is; operate too loaded down with trivial details; need separately to get test tube, be first numbered mark, then repeatedly centrifugal through at least twice.Though through two times centrifugal repeatedly, but for the macrobead more difficult removal in this way that proportion is similar to maxicell, in addition, easy generation operator in violation of rules and regulations, such as numbering error, causes the possibility of makeing mistakes to increase, and can not well be separated by macrobead, and therefore affect final experiment effect, thus derive a wrong conclusion.Above-mentioned two kinds of separation methods, all complex operations; And owing to needing during operation repeatedly repeatedly to get liquid liquid feeding, easily cause cell mechanical injuries and easily pollute, very strict to the environmental requirement in laboratory, thus affect cell quality, increase cellular segregation cost.
Meanwhile, in the cell separation apparatus of prior art, waste liquor recovery process is convenient not.
For this reason, need to provide a kind of better for the device of cellular segregation.
Summary of the invention
The object of the invention is to provide one can also collect required cell by sharp separation, easy and simple to handle and can alleviate sepn process produce cell injury, not easily cause cell contamination and easily carry out the cell separation apparatus of devil liquor recovery, this cell device can on one side feed liquor, carry out filtrations one side effluent discharge simultaneously, improve the efficiency of centrifugation.The present invention simultaneously also provides a kind of skimmer for this cell separation apparatus, and this skimmer makes waste collection process become simple, and operator directly just can collect waste liquid up, has not only saved the time, has avoided contaminate environment simultaneously.
Cell separation apparatus of the present invention, comprise vertical arrange main shaft, be connected to described main shaft and the centrifuge arm rotated around described main shaft, be fixedly connected on the cellular segregation collector of described centrifuge arm at least one end arranged transversely;
Wherein, the top of described main shaft is provided with skimmer, described skimmer comprise input duct, the admission chamber be connected with described input duct, fluid pipeline and be connected with described fluid pipeline go out sap cavity, described admission chamber and described go out sap cavity be provided with infusion port;
Described cellular segregation collector comprise the fluid inlet that is connected with the infusion port of described admission chamber with described go out the waste liquid chamber that is communicated with described liquid outlet of the acquisition cavity that is communicated with described fluid inlet of the liquid outlet that is connected of the infusion port of sap cavity and the filter membrane that is arranged between described acquisition cavity and described waste liquid chamber;
The diameter of the filter opening of described filter membrane is less than the diameter wanting isolated cell.
Further, described fluid inlet and described acquisition cavity are positioned at the far-end of described centrifuge arm, and described liquid outlet and described waste liquid chamber are positioned at the near-end of described centrifuge arm.
Further, described admission chamber and described go out sap cavity separate and arrange.
Preferably, the described input duct of described skimmer and described fluid pipeline are coaxially arranged.
Preferably, the described input duct of described skimmer and described fluid pipeline are arranged in parallel.
Preferably, described cell separation apparatus comprises two the cellular segregation collectors laying respectively at described centrifuge arm two ends, described admission chamber with described go out sap cavity be respectively provided with two infusion ports to be connected with described liquid outlet with the described fluid inlet of cellular segregation collector.
Preferably, the cross section of described cellular segregation collector is circular or square.
Preferably, the diameter of the filter opening of described filter membrane is 1-30um.
Preferably, described filter membrane is made up of polyethylene, cellulose mixture, PE material or nylon material.
The present invention also provides a kind of skimmer for cell separation apparatus, described cell separation apparatus comprises the cellular segregation collector with fluid inlet and liquid outlet, it is characterized in that: described skimmer comprise input duct, the admission chamber be connected with described input duct, fluid pipeline and be connected with described fluid pipeline go out sap cavity, described admission chamber and described go out sap cavity be provided with infusion port;
The infusion port of described admission chamber is communicated with the fluid inlet of described cellular segregation collector; Described go out the infusion port of sap cavity be communicated with the liquid outlet of described cellular segregation collector.
Preferably, in skimmer of the present invention, described admission chamber and described go out sap cavity separate and arrange.
Preferably, in skimmer of the present invention, described input duct and the described fluid pipeline of described skimmer are coaxially arranged.
Preferably, in skimmer of the present invention, described input duct and the described fluid pipeline of described skimmer are arranged in parallel.
Adopt cell separation apparatus of the present invention, structure is simple, easy to operate, can isolated cell rapidly and accurately, be separated and adopt dynamic centrifugal to be separated, be i.e. feed liquor body, filter liquide one side effluent discharge, whole filtration procedure carries out in totally-enclosed system, not easily cause damage and the pollution of cell, and isolated waste liquid is also easy to collect, refuse process is also easier.And on cell separation apparatus, adopt skimmer, make waste collection process become simple, operator directly just can collect waste liquid above cell separation apparatus, have not only saved the time, avoid contaminate environment simultaneously.
Accompanying drawing explanation
Fig. 1 is the one-piece construction schematic diagram of cell separation apparatus of the present invention;
Fig. 2 is the cross-sectional view of cell separation apparatus of the present invention;
Fig. 3 is the cross-sectional view of a kind of variation of cell separation apparatus of the present invention;
Fig. 4 is the structural representation of a kind of skimmer of cell separation apparatus of the present invention;
Fig. 5 is the structural representation of the another kind of skimmer of cell separation apparatus of the present invention.
Embodiment
Describe the embodiment of cell separation apparatus of the present invention in detail below in conjunction with accompanying drawing, but the present invention is not limited to these embodiments.
Embodiment one
Shown in Fig. 1, Fig. 2, Fig. 4, Fig. 5, cell separation apparatus of the present invention, comprises main shaft 1, is connected to main shaft 1 and the centrifuge arm 2 rotated around main shaft 1, is fixedly connected on the cellular segregation collector 3 of centrifuge arm 2 one end arranged transversely.
Skimmer 4 is provided with above the main shaft of cell separation apparatus, as shown in Figure 4,5, skimmer 4 comprise input duct 41, the admission chamber 43 be connected with input duct, fluid pipeline 42 and be connected with fluid pipeline go out sap cavity 44, admission chamber 43 with go out sap cavity 44 and be provided with infusion port.
As shown in Figure 2, cellular segregation collector 3 comprises the waste liquid chamber 34 that acquisition cavity 33 that liquid outlet 32 that the fluid inlet 31 that is connected with the infusion port 431 of admission chamber 43 is connected with the infusion port 441 going out sap cavity 44 is communicated with fluid inlet 31 is communicated with liquid outlet 32 and the filter membrane 35 be arranged between acquisition cavity 33 and waste liquid chamber 34.The diameter of the filter opening of filter membrane 35 is less than the diameter wanting isolated cell.
Further, skimmer 4 is positioned at directly over main shaft 1.
As shown in Figure 2, fluid inlet 31 and acquisition cavity 33 are positioned at the far-end of centrifuge arm 2, and liquid outlet 32 and waste liquid chamber 34 are positioned at the near-end of centrifuge arm 2, and that is to say that the position of acquisition cavity 33 is compared with the position in waste liquid chamber 34, the luxuriant central arm of acquisition cavity 33 2 is far away than waste liquid chamber 34.Like this, liquid flows to liquid outlet 32 from fluid inlet 31, is to flow from away from centrifuge arm 2 to nearly centrifuge arm 2 direction; Thus make when cell suspension arrives acquisition cavity 33, while cell is isolated in acquisition cavity 33 by filter membrane 35, acquisition cavity 33 can be fallen to away from filter membrane 35 under the influence of centrifugal force, thus can not filter opening be blocked, filtration procedure be continued smooth and easy.Moisture content and the waste liquid such as biological particles and biomolecules are then because liquid stream pressure enters waste liquid chamber 34 by filter membrane 35.
As shown in Figure 4,5, the admission chamber 43 of skimmer 4 with go out sap cavity 44 and be separated, such as can be separated by sealing-ring 40, can prevent liquid from discharging via liquid outlet when just not entered out sap cavity when abundant centrifuging like this, and can make liquid enter cellular segregation collector by abundant centrifuging after discharge again; Further due to admission chamber 43 with go out sap cavity 44 and be separated, liquid to be filtered with filter after waste liquid can not be confused, effluent discharge while can be implemented in feed liquor filter liquide.
The input duct 41 of skimmer 4 and fluid pipeline 42 both also can be able to be collocated (as shown in Figure 4) by arranged coaxial (as shown in Figure 5).When adopting arranged coaxial, skimmer can be saved and taken up space; When employing configures side by side, the structure that can be easier to realize skimmer also can be cost-saving.By arranging skimmer, making liquid feeding end and outlet end be positioned at homonymy, the taking back and process of waste liquid can be realized better.
After adopting skimmer 4, time centrifugal, liquid to be filtered enters the admission chamber 43 of cell separation apparatus from input duct 41, cellular segregation collector is entered from the infusion port 431 of admission chamber 43, under the effect of centrifugal force and liquid propulsive force, the waste liquid filtered out enters waste liquid chamber 34 through filter membrane 35, enter out the infusion port 441 of sap cavity 44 by liquid outlet 32 thus enter out sap cavity, waste liquid collecting bag is flowed into again by fluid pipeline 42, therefore skimmer 4 be arranged so that waste collection process becomes simple, operator directly just can collect waste liquid up, such mode also saves the time, avoid contaminate environment simultaneously.And target cell is stopped by filter membrane 35 and throws away filter membrane 35 in the effect of centrifugal force and fall to acquisition cavity 33, like this, terminate centrifugal after, operator directly can take cellular segregation collector away and obtain useful cell.
Filter membrane 35 is hydrophilic filter film, and it adopts polyolefins or polyamide polymer to make.Preferably, filter membrane is made up of polyethylene or cellulose mixture or PE material or nylon material.The filter opening of filter membrane 35 is less than the size of the cell wanting separation and Extraction, thus the cell that must extract is isolated be deposited in acquisition cavity 33, and moisture content and other biomolecules flow to waste liquid chamber 34, then flow out from liquid outlet 2.Because general somatic size is 5-30um, therefore preferably, the diameter of the filter opening of filter membrane is 1-30um; More preferably, the diameter of filter opening is 1um-5um.
Preferably, in the present embodiment, the acquisition cavity of cellular segregation collector 3 be there is inner chamber and have certain thickness shape, the inner chamber of such as acquisition cavity is towards the diminishing shape in direction away from filter membrane, such as back taper, and its cross section can be square or circular, adopt such structure, because the centrifugal force farthest away from centrifugal shaft position is maximum, cell can be made to be deposited on acquisition cavity better, make it be not easy blocking to filter membrane hole place.
The feed liquor face of filter membrane 35 is positioned at centrifuge arm far-end, goes out liquid level and is positioned at centrifuge arm near-end.Centrifuge arm rotates and produces centrifugal force, thus makes the cell that stopped by filter membrane be subject to greatly larger centrifugal force and " being got rid of " from filter membrane because of its particle, maintains filter opening unobstructed, moisture content and useless or harmful biomolecules are flowed away by filter opening under hydraulic action.
Further preferably, centrifuge arm 2 length can be 10-30cm, and rotating speed can be 500-2000 rev/min, and the centrifugal force of generation can be 50-1000g; More preferably, rotating speed can be 500-1500 rev/min, and the centrifugal force of generation can be 100-700g.
In centrifugal process, owing to have employed the structure of skimmer, make whole cell separation apparatus be totally enclosed, feed liquor and fluid have special pipeline, can not pollute target cell, operate also very easy; Further, centrifugal process is dynamic centrifugal, can realize feed liquor, filter liquide effluent discharge on one side.In addition, the admission chamber in skimmer with go out sap cavity and be separated, after can making liquid to be filtered and filtering, waste liquid is separated completely, there will not be liquid not also to be filtered the situation of just being discharged by fluid pipeline.Under the influence of centrifugal force, useful target cell constantly gathers and not easily blocks filter membrane hole in the acquisition cavity 33 of collector.And waste liquid flows into waste liquid collecting bag by the fluid pipeline of skimmer, such mode also makes waste liquid comparatively easily collecting.
Embodiment two
Shown in Figure 3, cell separation apparatus comprises two cellular segregation collector 3a and 3b laying respectively at centrifuge arm 2 two ends.And the admission chamber 43 of skimmer 4 with go out sap cavity 44 and be respectively provided with two infusion ports to be connected with liquid outlet with the fluid inlet of cellular segregation collector respectively.
Such as, an infusion port of admission chamber connects the fluid inlet of cellular segregation collector 3a, and the liquid outlet of cellular segregation collector 3a connects an infusion port of sap cavity.And another infusion port of admission chamber 43 connects the fluid inlet of cellular segregation collector 3b, the liquid outlet of cellular segregation collector 3b connects another infusion port of sap cavity 44.
In this embodiment, add cellular segregation collector, like this can disposable filtering more many cells suspension, further increase the working efficiency of apparatus of the present invention, save time and cost.
Content described in this specification sheets embodiment is only enumerating the way of realization of inventive concept; protection scope of the present invention should not be regarded as being only limitted to the specific form that embodiment is stated, protection scope of the present invention also and conceive the equivalent technologies means that can expect according to the present invention in those skilled in the art.
Claims (13)
1. a cell separation apparatus, is characterized in that: comprise vertical arrange main shaft, be connected to described main shaft and the centrifuge arm rotated around described main shaft, be fixedly connected on the cellular segregation collector of described centrifuge arm at least one end arranged transversely;
Wherein, the top of described main shaft is provided with skimmer, described skimmer comprise input duct, the admission chamber be connected with described input duct, fluid pipeline and be connected with described fluid pipeline go out sap cavity, described admission chamber and described go out sap cavity be provided with infusion port;
Described cellular segregation collector comprise the fluid inlet that is connected with the infusion port of described admission chamber with described go out the waste liquid chamber that is communicated with described liquid outlet of the acquisition cavity that is communicated with described fluid inlet of the liquid outlet that is connected of the infusion port of sap cavity and the filter membrane that is arranged between described acquisition cavity and described waste liquid chamber;
The diameter of the filter opening of described filter membrane is less than the diameter wanting isolated cell.
2. cell separation apparatus as claimed in claim 1, it is characterized in that: described fluid inlet and described acquisition cavity are positioned at the far-end of described centrifuge arm, described liquid outlet and described waste liquid chamber are positioned at the near-end of described centrifuge arm.
3. cell separation apparatus as claimed in claim 1, is characterized in that: described admission chamber and described go out sap cavity separate and arrange.
4. cell separation apparatus as claimed in claim 1, is characterized in that: described input duct and the described fluid pipeline of described skimmer are coaxially arranged.
5. cell separation apparatus as claimed in claim 1, is characterized in that: described input duct and the described fluid pipeline of described skimmer are arranged in parallel.
6. the cell separation apparatus according to any one of claim 1-5, it is characterized in that: described cell separation apparatus comprises two the cellular segregation collectors laying respectively at described centrifuge arm two ends, described admission chamber with described go out sap cavity be respectively provided with two infusion ports to be connected with described liquid outlet with the described fluid inlet of cellular segregation collector.
7. cell separation apparatus as claimed in claim 1, is characterized in that: the cross section of described cellular segregation collector is for circular or square.
8. cell separation apparatus as claimed in claim 1, is characterized in that: the diameter of the filter opening of described filter membrane is 1-30um.
9. cell separation apparatus as claimed in claim 1, is characterized in that: described filter membrane is made up of polyethylene, cellulose mixture, PE material or nylon material.
10. the skimmer for cell separation apparatus, described cell separation apparatus comprises the cellular segregation collector with fluid inlet and liquid outlet, it is characterized in that: described skimmer comprise input duct, the admission chamber be connected with described input duct, fluid pipeline and be connected with described fluid pipeline go out sap cavity, described admission chamber and described go out sap cavity be provided with infusion port;
The infusion port of described admission chamber is communicated with the fluid inlet of described cellular segregation collector; Described go out the infusion port of sap cavity be communicated with the liquid outlet of described cellular segregation collector.
11., as claimed in claim 10 for the skimmers of cell separation apparatus, is characterized in that: described admission chamber and described go out sap cavity separate and arrange.
12., as claimed in claim 10 for the skimmer of cell separation apparatus, is characterized in that: described input duct and the described fluid pipeline of described skimmer are coaxially arranged.
13., as claimed in claim 10 for the skimmer of cell separation apparatus, is characterized in that: described input duct and the described fluid pipeline of described skimmer are arranged in parallel.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510560187.5A CN105112289A (en) | 2015-09-02 | 2015-09-02 | Cell separation device and dispenser applied to same |
| US15/230,516 US10258927B2 (en) | 2012-12-14 | 2016-08-08 | Centrifugal dynamic filtering apparatus and cell separation system using same |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201510560187.5A CN105112289A (en) | 2015-09-02 | 2015-09-02 | Cell separation device and dispenser applied to same |
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| CN105112289A true CN105112289A (en) | 2015-12-02 |
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| Application Number | Title | Priority Date | Filing Date |
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| CN201510560187.5A Pending CN105112289A (en) | 2012-12-14 | 2015-09-02 | Cell separation device and dispenser applied to same |
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| CN (1) | CN105112289A (en) |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN108187923A (en) * | 2018-01-04 | 2018-06-22 | 北京市春立正达医疗器械股份有限公司 | A kind of centrifugal blood component for centrifuge |
| CN109628287A (en) * | 2018-12-28 | 2019-04-16 | 科兴（大连）疫苗技术有限公司 | Cell collects shaking table and cell collection method |
| CN109913350A (en) * | 2019-03-22 | 2019-06-21 | 李闯 | Integrated device for detecting and extracting tumor cells |
| CN110079439A (en) * | 2019-05-15 | 2019-08-02 | 林伟阳 | A kind of cell isolation method based on two-freedom continuous centrifuge |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN109628287A (en) * | 2018-12-28 | 2019-04-16 | 科兴（大连）疫苗技术有限公司 | Cell collects shaking table and cell collection method |
| CN109913350A (en) * | 2019-03-22 | 2019-06-21 | 李闯 | Integrated device for detecting and extracting tumor cells |
| CN110079439A (en) * | 2019-05-15 | 2019-08-02 | 林伟阳 | A kind of cell isolation method based on two-freedom continuous centrifuge |
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Application publication date: 20151202 |