CN103243026B - Multifunctional full-automatic cell and solution treating instrument - Google Patents
Multifunctional full-automatic cell and solution treating instrument Download PDFInfo
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- CN103243026B CN103243026B CN201210032654.3A CN201210032654A CN103243026B CN 103243026 B CN103243026 B CN 103243026B CN 201210032654 A CN201210032654 A CN 201210032654A CN 103243026 B CN103243026 B CN 103243026B
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Abstract
The invention relates to a medical instrument, and especially relates to a multifunctional full-automatic cell and solution treating instrument. The instrument comprises an ultrafilter, a liquid storage container, a peristaltic pump, an electromagnetic valve mixer and a temperature controller, wherein special components in a cell suspension or solution are displaced by a method of instillation combined with ultrafiltration and dialysis, or the cell suspension or solution is diluted and concentrated to desired final volume through accurate control. The multifunctional full-automatic cell and solution treating instrument effectively overcomes the shortages of the existing related treatment methods in the aspects of causing damage of cells, possibly causing cell pollution, taking long time, and low efficiency and the like; and the multifunctional full-automatic cell and solution treating instrument is capable of adding or removing a low-temperature protector quickly and steadily without pollution, and also capable of controlling the volumes and the temperatures of the cell suspension and the solution quite accurately and reducing various cell damages possibly caused in the treatment process. The multifunctional full-automatic cell and solution treating instrument has the advantages of small volume, simple structure, little time consumed and high controllability, and is convenient to operate.
Description
Technical field
The present invention relates to a kind of Medical Instruments, particularly the special component in a kind of displacement of the method by perfusion, ultrafiltration and dialysis cell suspending liquid or multi-functional full-automatic cell and the solution-treated instrument that cell suspending liquid is diluted and concentrated
Background technology
In the low temperature or process of cryopreservation of cell, in order to reduce low temperature injury, often need in the cell suspending liquid being saved, add the cryoprotective agent of particular types and certain concentration.For example, preserving the conventional cryoprotective agent of mankind hemopoietic stem cell is dimethyl sulfoxide (DMSO) (DMSO), and what preserve that human erythrocyte commonly uses is glycerine.And in therapeutic process; if not scavenger cell suspension and intracellular cryoprotective agent in advance; and directly by the cell suspending liquid input human body that contains cryoprotective agent; body inner blood will cause osmotic pressure damage to the cell of input to the rapid dilution of input cell suspending liquid, may cause the failure of Transplanted cells.In addition, the cryoprotective agent of these input human bodies is at room temperature to human body toxic side effect.Therefore, after rewarming, from cell suspending liquid, remove cryoprotective agent very necessary.
In the research process such as stem cell, cell cultures gained cell suspension has the features such as volume large (reaching several liters), cell concn are low conventionally.For the ease of follow-up applied research, generally need to concentrated hundreds if not thousands of times of gained cell suspension will be cultivated.In addition, some otherly to the dilution of cell suspension or to the displacement of solution composition, be also common in various clinical and research application.
At present conventional associated processing method is the method based on centrifugal (as the stepwise dilution of removing in cryoprotective agent process adds centrifuging).In this method, the mechanical force and the osmotic pressure that due to high speed centrifugation rotation, produce often change and can bring serious physical abuse and osmotic pressure damage to cell; And because centrifugal separation processes is carried out in open environment, this operational misclosure also can cause living material to be subject to the pollution of bacterium; In addition, centrifuging complex operation, inefficiency, processing a cell suspending liquid needs 2-4 hour conventionally.
Summary of the invention
The object of this invention is to provide a kind of effectively overcome existing associated processing method causing cell injury, may cause the deficiency of the aspects such as cell contamination and length consuming time, inefficiency; can quick and stable contamination-freely add or remove cryoprotective agent; and can more accurately control volume and the temperature of adding or removing speed and cell suspending liquid, reduce the interpolation of the various cell injury that may cause in treating processes and remove multi-functional full-automatic cell and the solution-treated instrument of cryoprotective agent in cell suspending liquid.
The technical scheme that realizes the object of the invention is: a kind of multi-functional full-automatic cell and solution-treated instrument, there is ultra-fine filter, cell suspension liquid container, displacement liquid container and waste fluid container, between each container and ultra-fine filter, by tubing, be connected, wherein cell suspension liquid container is by the inner chamber import UNICOM of the first tubing and ultra-fine filter, inner chamber outlet UNICOM by the second tubing and ultra-fine filter forms cell suspending liquid loop simultaneously, displacement liquid container is by the exocoel import UNICOM of the 3rd tubing and ultra-fine filter, waste fluid container is by the exocoel outlet UNICOM of the 4th tubing and ultra-fine filter, described cell suspending liquid loop is provided with first sensor and the first peristaltic pump, the 3rd tubing is provided with the second sensor, the second peristaltic pump, temperature controller and dialysis magnetic valve, the 4th tubing is provided with the 3rd sensor and the 3rd peristaltic pump, and be respectively equipped with at least one mixing tank on the first tubing and the second tubing, each mixing tank is by one section of tubing and corresponding magnetic valve and San tubing UNICOM.
The mixing tank of above-mentioned multi-functional full-automatic cell and solution-treated instrument has and a plurality ofly (as: has five, be respectively series connection and be arranged on the 4th mixing tank and the 5th mixing tank on the first tubing, and series connection is arranged on the first mixing tank on the second tubing, the second mixing tank and the 3rd mixing tank, wherein the first mixing tank is connected with the first magnetic valve by the first mixing tubing, the second mixing tank is connected with the second magnetic valve by the second mixing tubing, the 3rd mixing tank mixes tubing by the 3rd and is connected with the 3rd magnetic valve, the 4th mixing tank mixes tubing by the 4th and is connected with the 4th magnetic valve, the 5th mixing tank mixes tubing by the 5th and is connected with the 5th magnetic valve).
Each peristaltic pump, magnetic valve, sensor and the temperature controller of above-mentioned multi-functional full-automatic cell and solution-treated instrument are all connected with main control computer, main control computer is the feedback signal formation instruction in conjunction with each sensor according to selected handling procedure, control state and the operating parameter of temperature controller, each peristaltic pump, magnetic valve, thereby control cell suspension and flow velocity, the flow direction and the temperature of other solution in consumptive material.
The dialysis magnetic valve of above-mentioned multi-functional full-automatic cell and solution-treated instrument, the first magnetic valve, the second magnetic valve, the 3rd magnetic valve, the 4th magnetic valve and the 5th magnetic valve are pinch valve.
The tubing of above-mentioned multi-functional full-automatic cell and solution-treated instrument is one-time infusion tube.
The sensor of above-mentioned multi-functional full-automatic cell and solution-treated instrument comprises that temperature sensor and electricity lead sensing device, temperature sensor measurement temperature signal feeds back to main control computer, and control temperature controller by main control computer, electricity is led sensing device by the electric conductivity value of measurements and calculations control cell suspension, realize solute concentration in monitoring cell suspension and change, real-time judge is processed progress.
The present invention has positive effect:
(1) multi-functional full-automatic cell of the present invention and solution-treated instrument can add or remove cryoprotective agent in cell suspending liquid efficiently, or cell suspending liquid is concentrated or diluted, can effectively overcome existing method causing cell injury, may cause the deficiency of the aspects such as cell contamination and length consuming time, inefficiency, can quick and stable contamination-freely add or remove cryoprotective agent, and can more accurately control volume and the temperature of cell suspending liquid, reduce the various cell injury that may cause in treating processes;
(2) in the present invention, magnetic valve used is pinch valve, pipeline is through valve body, by flexible pipe in the contraction pilot piping of magnetic valve, be unimpeded or close, itself does not directly contact with the medium in pipeline, be used in conjunction with and can guarantee cleaning system sealing with peristaltic pump, avoid enchylema and scavenging solution bacterial contamination;
(3) in the present invention, adopt thermostat, by controlling displacement liquid temperature, reach on the one hand the object of final control blood heat, on the other hand, improve the temperature of displacement liquid and blood and can strengthen diffusion effect, quickening displacement liquid mixes with blood, or improves the efficiency of cleaning;
(4) to have volume little, simple in structure, easy to operate, consuming time few in the present invention, the advantage that controllability is strong.
Accompanying drawing explanation
For content of the present invention is more easily expressly understood, according to specific embodiment also by reference to the accompanying drawings, the present invention is further detailed explanation, wherein below
Fig. 1 is structural representation of the present invention.
Embodiment
See Fig. 1, the present invention has ultra-fine filter 3, cell suspending liquid container C 1, displacement liquid container C 2 and waste fluid container C3, ultra-fine filter 3 adopts hollow fiber type ultra-fine filter, there is inner chamber import 31, inner chamber outlet 32, exocoel import 33 and exocoel outlet 34, cell suspending liquid container C 1, between displacement liquid container C 2 and waste fluid container C3 and ultra-fine filter 3, by tubing, be connected, wherein cell suspending liquid container C 1 is by inner chamber import 31 UNICOMs of the first tubing 61 and ultra-fine filter 3, the second tubing 62 exports 32 UNICOMs' formation loops with the inner chamber of ultra-fine filter 3, displacement liquid container C 2 is exocoel import 33 UNICOMs with ultra-fine filter 3 by the 3rd tubing 63, waste fluid container C3 exports 34 UNICOMs by the 4th tubing 64 and the exocoel of ultra-fine filter 3, described cell suspending liquid container C 1 is provided with first sensor 21 and the first peristaltic pump P1 with ultra-fine filter 3 loops, the 3rd tubing 63 is provided with the second sensor 22, the second peristaltic pump P2, temperature controller 1 and dialysis magnetic valve V, the 4th tubing 64 is provided with the 3rd sensor 23 and the 3rd peristaltic pump P3, and be respectively equipped with five mixing tanks on the first tubing 61 and the second tubing 62, each mixing tank is by a corresponding magnetic valve and the 3rd tubing 63 UNICOMs, five mixing tanks are respectively series connection and are arranged on the 4th mixing tank 44 and the 5th mixing tank 55 on the first tubing 61, and series connection is arranged on the first mixing tank 41 on the second tubing 62, the second mixing tank 42 and the 3rd mixing tank 43, wherein the first mixing tank 41 is connected with the first magnetic valve V1 by the first mixing tubing 631, the second mixing tank 42 is connected with the second magnetic valve V2 by the second mixing tubing 632, the 3rd mixing tank 43 mixes tubing 633 by the 3rd and is connected with the 3rd magnetic valve V3, the 4th mixing tank 44 mixes tubing 634 by the 4th and is connected with the 4th magnetic valve V4, the 5th mixing tank 45 mixes tubing 635 by the 5th and is connected with the 5th magnetic valve V5, each peristaltic pump, magnetic valve, sensor and temperature controller 1 are all connected with Controlling System, sensor comprises that temperature sensor and electricity lead sensing device, temperature sensor measurement temperature signal feeds back to Controlling System, and control temperature controller 1 by Controlling System, electricity is led sensing device by the electric conductivity value of measurements and calculations control cell suspension or waste liquid, realizing solute concentration in monitoring cell suspension changes, real-time judge is processed progress, Controlling System is the feedback signal formation instruction in conjunction with each sensor according to selected handling procedure, control temperature controller 1, each peristaltic pump, the state of magnetic valve and operating parameter, thereby control cell suspension and the flow velocity of other solution in consumptive material, flow to and temperature, magnetic valve V dialyses in the present embodiment, the first magnetic valve V1, the second magnetic valve V2, the 3rd magnetic valve V3, the 4th magnetic valve V4 and the 5th magnetic valve V5 are pinch valve, tubing is one-time infusion tube, ultra-fine filter 3 also can adopt during high flux hemodialysis device to replace.
The principles of science of foundation of the present invention is: (1) turbulent flow is forced mixing: the cell suspending liquid in consumptive material and displacement liquid one or more mixing tanks of flowing through under host driven; carry out single-point or multiple spot perfusion; and under mixing tank effect, form strong turbulent flow; to realize higher mass transfer efficiency; thereby promote the Quick uniform of cell suspending liquid and displacement liquid to mix, the cryoprotective agent in displacement liquid enters rapidly cryoprotective agent in cell suspending liquid (adding procedure) or cell suspending liquid by rapid dilution (cleaning process).(2) filter, ultrafiltration principle: the cell suspending liquid ultra-fine filter of flowing through, a large amount of liquid movements that produce by the pressure difference by semi-permeable membranes both sides, will form the filtrate eliminating by water or cryoprotective agent by semi-permeable membranes in cell suspending liquid.(3) dialysis principle: cell suspending liquid is along a direction equally distributed porous hollow fiber semi-permeable membranes pipe vertically of flowing through in ultra-fine filter, meanwhile, the region of displacement liquid reverse flow outside semi-permeable membranes pipe.The concentration gradient that can produce cryoprotective agent at internal surface and the outside surface of semi-permeable membranes pipe like this, cryoprotective agent can be flowed to lower concentration direction by high density in the driving of this concentration gradient.Therefore by applying different displacement liquids, can reach respectively interpolation and remove protectant object.
Actual magnetic valve and the visual application requiring of mixing tank of using suitably changes, perfusion before forming respectively (i.e. " first ultrafiltration after perfusion ", an above perfusion point) or rear perfusion (" pouring into " an above perfusion point after first ultrafiltration).If shut electromagnetic valve valve V1, V2, V3, V4, realize perfusion before single-point, as shut electromagnetic valve V3, V4, V5, realize at 2 after perfusion.With respect to single-point perfusion, multiple spot reperfusion structure is more complicated, but is conducive to realize less Premeabilisation of cells crushing wound or allow larger perfusion dosage.First after perfusion, after ultrafiltration and first ultrafiltration, fill-up mode is distinguished the relative position that is mixing tank and ultra-fine filter 3.In parameter, arrange when identical, under " front perfusion " pattern, cell has better trafficability characteristic in ultra-fine filter 3, but " perfusion afterwards " pattern has better processing efficiency, therefore when high appropriate to the occasion selection " front perfusion " pattern of cell concn, and when cell concn is low, select " perfusion afterwards " pattern.
The effect of the first peristaltic pump P1 is cell suspending liquid to be entered to the inner chamber import 31 of ultra-fine filter 3 by desired flow pump, the effect of the second peristaltic pump P2 is that the effect that displacement liquid is pumped into exocoel import 33, the three peristaltic pump P3 of mixing tank or ultra-fine filter 3 by desired flow velocity is the flow velocity of controlling the exocoel of ultra-fine filter 3 outlet 34.The first peristaltic pump P1 and the second peristaltic pump P2 associated working can realize the Dilution ratio of cell suspending liquid in mixing tank, and the 3rd peristaltic pump P3 can realize Filtration Control.
Below just to take the cryoprotective agent cleaning in red blood cell suspension be example, further illustrate:
The first step, ultrafiltration is cleaned: the first magnetic valve V1, the second magnetic valve V2, the 3rd magnetic valve V3, the 4th magnetic valve V4 and the 5th magnetic valve V5 open, dialysis magnetic valve V closes, the first peristaltic pump P1, the second peristaltic pump P2 and the 3rd peristaltic pump P3 open, cell suspending liquid is through mixing tank and displacement liquid mixed diluting is laggard enters ultra-fine filter 3, part ultrafiltration forms waste liquid and enters waste fluid container C3, and remainder returns to cell suspending liquid container C 1, forms circulation.Wherein, the rotating speed of the second peristaltic pump P2 constantly adjusts to keep higher cleaning efficiency with cleaning process; The 3rd peristaltic pump P3 rotating speed divides third gear: acquiescence D, high H and low L, and acquiescence rotating speed D is consistent with the second peristaltic pump P2.First sensor 21, the second sensor 22 and the 3rd sensor 23 weigh displacement liquid consumption and ultrafiltrated growing amount---that is the groundwater increment of cell suspending liquid and the contrast situation of ultrafiltration volume, and information is fed back to Controlling System, adjust the 3rd peristaltic pump P3 rotating speed: when ultrafiltration volume is greater than groundwater increment, adjusting the 3rd peristaltic pump P3 rotating speed is low L, when ultrafiltration volume is less than groundwater increment, adjusting the 3rd peristaltic pump P3 rotating speed is high H, thereby realize, keeps cell suspending liquid volumetric balance in ultrafiltration cleaning process.
Second step; dialysis is cleaned: the first magnetic valve V1 to the five magnetic valve V5 are closed; dialysis magnetic valve V opens; the exocoel import 33 that allows displacement liquid enter ultra-fine filter 3 as dialyzate by the 3rd tubing 63---temperature controller 1---dialysis magnetic valve V---tubing 63 '; and enter waste fluid container C3 by exocoel outlet 34; blood enters ultra-fine filter 3 by the first peristaltic pump P1 recycle pump simultaneously; form a cell suspending liquid and isotonic saline solution in the countercurrent flow of hollow-fibre membrane both sides; cryoprotective agent in cell suspending liquid is constantly eliminated, until reach removing requirement.
The 3rd step, ultrafiltration and concentration: the first peristaltic pump P1, the 3rd peristaltic pump P3 operation, the second peristaltic pump P2 stops, magnetic valve Close All.Cell suspending liquid is concentrated under Ultrafiltration, until meet the requirements of cell pack ratio.
Above-described specific embodiment; object of the present invention, technical scheme and beneficial effect are further described; institute is understood that; the foregoing is only specific embodiments of the invention; be not limited to the present invention; within the spirit and principles in the present invention all, any modification of making, be equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (5)
1. a multi-functional full-automatic cell and solution-treated instrument, there is ultra-fine filter (3), cell suspension liquid container (C1), displacement liquid container (C2) and waste fluid container (C3), between each container and ultra-fine filter (3), by tubing, be connected, wherein cell suspension liquid container (C1) is by inner chamber import (31) UNICOM of the first tubing (61) and ultra-fine filter (3), by the second tubing (62), form cell suspending liquid loop with inner chamber outlet (32) UNICOM of ultra-fine filter (3) simultaneously, displacement liquid container (C2) is exocoel import (33) UNICOM with ultra-fine filter (3) by the 3rd tubing (63), waste fluid container (C3) is exocoel outlet (34) UNICOM with ultra-fine filter (3) by the 4th tubing (64), it is characterized in that: described cell suspension liquid container (C1) is provided with first sensor (21) and the first peristaltic pump (P1) with ultra-fine filter (3) loop, the 3rd tubing (63) is provided with the second sensor (22), the second peristaltic pump (P2), temperature controller (1) and dialysis magnetic valve (V), the 4th tubing (64) is provided with the 3rd sensor (23) and the 3rd peristaltic pump (P3), and be respectively equipped with at least one mixing tank on the first tubing (61) and the second tubing (62), each mixing tank is by one section of tubing and corresponding magnetic valve and the 3rd tubing (63) UNICOM, described mixing tank has five, be respectively series connection and be arranged on the 4th mixing tank (44) and the 5th mixing tank (55) on the first tubing (61), and series connection is arranged on the first mixing tank (41) on the second tubing (62), the second mixing tank (42) and the 3rd mixing tank (43), wherein the first mixing tank (41) is connected with the first magnetic valve (V1) by the first mixing tubing (631), the second mixing tank (42) mixes tubing (632) by second and is connected with the second magnetic valve (V2), the 3rd mixing tank (43) mixes tubing (633) by the 3rd and is connected with the 3rd magnetic valve (V3), the 4th mixing tank (44) mixes tubing (634) by the 4th and is connected with the 4th magnetic valve (V4), the 5th mixing tank (45) mixes tubing (635) by the 5th and is connected with the 5th magnetic valve (V5).
2. multi-functional full-automatic cell according to claim 1 and solution-treated instrument, it is characterized in that: described each peristaltic pump, magnetic valve, sensor and temperature controller (1) are all connected with Controlling System, Controlling System is the feedback signal formation instruction in conjunction with each sensor according to selected handling procedure, control state and the operating parameter of temperature controller (1), each peristaltic pump, magnetic valve, thereby control cell suspension and flow velocity, the flow direction and the temperature of other solution in consumptive material.
3. multi-functional full-automatic cell according to claim 1 and solution-treated instrument, is characterized in that: described dialysis magnetic valve (V), the first magnetic valve (V1), the second magnetic valve (V2), the 3rd magnetic valve (V3), the 4th magnetic valve (V4) and the 5th magnetic valve (V5) are pinch valve.
4. multi-functional full-automatic cell according to claim 1 and solution-treated instrument, is characterized in that: described tubing is one-time infusion tube.
5. multi-functional full-automatic cell according to claim 2 and solution-treated instrument, it is characterized in that: described sensor comprises that temperature sensor and electricity lead sensing device, temperature sensor measurement temperature signal feeds back to Controlling System, and control temperature controller (1) by Controlling System, electricity is led sensing device by the electric conductivity value of measurements and calculations cell suspension, realize solute concentration in monitoring cell suspension and change, real-time judge is processed progress.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210032654.3A CN103243026B (en) | 2012-02-14 | 2012-02-14 | Multifunctional full-automatic cell and solution treating instrument |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201210032654.3A CN103243026B (en) | 2012-02-14 | 2012-02-14 | Multifunctional full-automatic cell and solution treating instrument |
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| CN103243026A CN103243026A (en) | 2013-08-14 |
| CN103243026B true CN103243026B (en) | 2014-09-03 |
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Families Citing this family (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103725599B (en) * | 2014-01-14 | 2015-05-27 | 刘韬 | Fluid film filter cell separation device |
| CN104611218B (en) * | 2015-01-29 | 2017-02-22 | 中国科学技术大学 | Device and method for removing low-temperature protective agent in cell suspension solution |
| EP3093335A1 (en) * | 2015-05-13 | 2016-11-16 | Bayer Technology Services GmbH | Process control system for regulating and controlling a modular system for the production of biopharmaceutical and biological macromolecular products |
| CN105925475B (en) * | 2016-06-16 | 2018-02-09 | 芜湖中科医凌生命科技有限公司 | A kind of full-automatic concentration instrument |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN201046965Y (en) * | 2007-05-14 | 2008-04-16 | 高大勇 | Universal cell processor for adding and removing low-temperature protecting agent in cell suspension |
| CN101173209A (en) * | 2006-11-02 | 2008-05-07 | 杭州晶格科学仪器有限公司 | Diaphragm type liquid basal cell gathering system and method thereof |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| DE102004023080B4 (en) * | 2004-05-11 | 2009-01-15 | Fresenius Medical Care Deutschland Gmbh | Method and device for monitoring the supply of substitution fluid during extracorporeal blood treatment |
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Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101173209A (en) * | 2006-11-02 | 2008-05-07 | 杭州晶格科学仪器有限公司 | Diaphragm type liquid basal cell gathering system and method thereof |
| CN201046965Y (en) * | 2007-05-14 | 2008-04-16 | 高大勇 | Universal cell processor for adding and removing low-temperature protecting agent in cell suspension |
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