CN101732771A - Cell reactor and artificial liver support system comprising same - Google Patents
Cell reactor and artificial liver support system comprising same Download PDFInfo
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- CN101732771A CN101732771A CN200910242907A CN200910242907A CN101732771A CN 101732771 A CN101732771 A CN 101732771A CN 200910242907 A CN200910242907 A CN 200910242907A CN 200910242907 A CN200910242907 A CN 200910242907A CN 101732771 A CN101732771 A CN 101732771A
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Abstract
The invention provides a cell reactor and an artificial liver support system comprising the same. The cell reactor comprises a container, a liquid outlet is formed at the bottom of the container, a sealing cover is arranged at the top of the container, a liquid inlet is formed on the sealing cover, layers of filter meshes of 100-400 meshes are respectively arranged at the bottom of the container and in the sealing cover at the top of the container, covers which are in seal fit with the liquid outlet and the inlet are formed at the liquid outlet and the inlet, and a mixture of a micro-carrier growing liver cells and resin is accommodated between the two layers of the filter meshes in the container. The reactor is connected on a dialysate circulation path of a blood purifying device for constituting the artificial liver support system, thereby leading the liver cells not to be in direct contact with blood and realizing good biological safety; furthermore, the reactor can be used for a plurality of times through regeneration treatment, thereby greatly reducing the treatment cost of a biological artificial liver.
Description
Technical field
The invention belongs to field of biomedicine technology, be specifically related to a kind of cell reactor and preparation method thereof and renovation process, also relate to the artificial liver support system that comprises this cell reactor.
Background technology
Liver failure is the whole performance in latter stage of various hepatopathys, its incidence rate and mortality rate are very high, artificial liver support system is present more convenient effective treatment means, and it can be the patient liver function Supporting Therapy is provided, thereby makes the patient successfully carry out the transition to liver transplantation or self impaired liver function recovery.Artificial liver comprises three types: abiotic type artificial liver, Biotype artificial liver and hybrid artificial liver.Wherein abiotic type artificial liver comprises hemodialysis, plasmapheresis, activated carbon adsorption etc., though can play certain Detoxication, differs greatly with real liver function.And the Biotype artificial liver is owing to use growth to have hepatocellular cell reactor that patient's blood plasma is detoxified and metabolism, its more perfect functions, but a large amount of toxicants is difficult to remove in a short time in the blood samples of patients, can produce damage by the interior hepatocyte of pair cell reactor.Hybrid artificial liver system synthesis the advantage of preceding two kinds of artificial livers, earlier blood/plasma is circulated by abiotic liver, reduce the concentration of noxious substance in the blood, and then by biological liver detoxify, metabolism, synthetic and secretion, the effect of its alternative liver is best, therefore has a good application prospect.
Real performance liver function is biological liver cyclic part in hybrid artificial liver system and the Biotype artificial liver system, but biological liver circulating device type is various, does not still have preferred plan at present.Because the major part that biological liver cyclic part is an artificial liver, therefore the improvement to this part is the emphasis of artificial liver development.At present, big quantity research has been carried out to cell material and the cell reactor type used among the bioartificial liver in the research center of a plurality of countries in the world, and existing part biological artificial liver system enters clinical experimental stage, and has obtained certain curative effect.And domestic still do not have at present can be applicable to clinical bioartificial liver and device thereof.Cell reactor is as the nucleus of bioartificial liver system, and not only directly cell activity and function are wherein loaded in influence, and directly influence the safety of bioartificial liver system.Before the present invention, the cell reactor of clinical use roughly is divided into 4 types: hollow fiber type, and the single flat template, perfusion bed or support rack type and microcapsule bag are by floating type.Wherein the hollow fiber type cell reactor is that application is maximum at present, it is that a branch of hollow fiber conduit is sealed in the shell of transparent organic material, this fiber is divided into inside and outside two chambeies with shell, the mobile patient's of general inner chamber blood plasma, and exocoel can install the liver cell of cultivation.This fiber also has the semipermeable membrane function simultaneously, realizes the two-way transportation of material in the time of isolated cell, thereby reaches the purpose of dialysis and function of detoxification.
Yet there is certain defective in this reactor in bioartificial liver's preparation.At first, the doughnut dense arrangement in this reactor makes the outer capacity of pipe and effectively exchange area is very little, is difficult for loading a large amount of hepatocyte; Secondly, cell can not be uniformly distributed between the doughnut after hepatocyte is packed into, causes blood plasma to reduce from acellular space by the detoxifcation efficient that makes cell; The 3rd, the easy dense accumulation of the hepatocyte of packing into makes the exchange of cytotrophy composition limited, has reduced hepatocellular activity and function.
In theory, ideal cell reactor should possess following condition: 1, doughnut has outward and holds enough hepatocellular space, makes a cell reactor can satisfy treatment to 1 routine liver failure patient.2, the microenvironment of hepatocyte survival and function can be provided.3, micromolecule passes through the two-way transportation of doughnut in the permission, and intercepts the macromolecular substances exchange.4. prolong the time that cell reactor is applied to treat as far as possible, the use of maybe can regenerating.Yet the cell reactor that does not up to the present also meet these conditions in the world is applied to clinical.
Summary of the invention
A technical problem to be solved by this invention provides a kind of cell reactor, and this cell reactor has been avoided in the hollow fiber bioreactor problem of uneven distribution behind the narrow and small regulating liver-QI cell device of the doughnut external space.Can also solving hepatocyte, directly to cultivate in bioreactor hepatic cell growth bad and be difficult to observe the problem of hepatic cell growth situation.
In order to solve the problems of the technologies described above, technical scheme provided by the present invention is:
The invention provides a kind of cell reactor, described cell reactor comprises a container, this container bottom is provided with liquid outlet, container top is provided with capping, capping is provided with the liquid inlet, be respectively equipped with one deck 100～400 purpose filter screens in container bottom and the container top capping, liquid outlet and porch are provided with the lid that is sealed and matched with it, hold the long mixture that hepatocellular microcarrier and resin are arranged between the two-layer filter screen of internal tank.
Load the long mixture that hepatocellular microcarrier and resin are arranged in the cell reactor of the present invention, wherein said hepatocyte is animal or human's primary hepatocyte, the human or animal's hepatic cell line or the tumor cell of liver of immortalization, described resin particle diameter is 100～4000 μ m, resin accounts for and loads 5～30% of cumulative volume, and it is 3: 1～9: 1 that described length has the volume ratio of hepatocellular microcarrier and mixed with resin.The purpose of mixing resin is liquid and the hepatocellular contact area that flows through reactor for increase, because the diameter of the microcarrier of cell culture is generally about 50～200 μ m, the resin diameter that the present invention uses is bigger 2～20 times than microcarrier, when with microcarrier with than the big mixed with resin of its diameter after, can there be a lot of holes in the mixture, hepatocellularly on the liquid that therefore can guarantee to flow through cell reactor and the microcarrier fully contact.Used resin particle can be the resin particle of any medical resin material preparation, and its concrete material is including, but not limited to polrvinyl chloride, polyethylene, polypropylene, low density polyethylene (LDPE), ultra-high molecular weight polyethylene, politef, FEP, polymethyl methacrylate, polyoxyethylene methylene, polyethylene terephthalate, polyurethane, silicone rubber, polysulfones, polylactic acid, polyglycolic acid etc.
Long have hepatocellular microcarrier and mixed with resin loading reactor to have tangible advantage: the one, and hepatic cell growth is on microcarrier, the hepatocyte density of cultivation is increased greatly, and hepatocyte can keep surviving and the transfer and the loading of excellent function state and very convenient cell for a long time; The 2nd, mix the bigger mixed with resin of diameter, make laden microcarrier keep certain liquid communication duct on every side, avoided the perfusion dead space, guaranteed that liquid fully contacts with hepatocellular; If the resin of Shi Yonging is an adsorbent resin in addition, also has the effect of absorbing toxin.
Another feature of cell reactor of the present invention is to be easy to amplify, and a plurality of cell reactor series connection of the present invention can be used, to strengthen its function.
Another technical problem to be solved by this invention provides a kind of preparation method of cell reactor.Its technical scheme is:
A kind of preparation method of above-mentioned cell reactor.At first hepatocyte fresh separated or that amplification in vitro is cultivated is seeded on the microcarrier, according to hepatic cell line 1 * 10
7～1 * 10
8The mixed of cell: 1～2g microcarrier: 500ml～1L culture medium is cultivated under the situation of shaking, and cell is attached on the microcarrier.Wherein said hepatocyte can be any hepatocyte that is applicable to the artificial liver treatment, and described microcarrier can be any microcarrier that is suitable for cell culture, comprises various solid microcarriers and various hollow microcarrier.Treat hepatocyte grow on the microcarrier 80～100% converge after, the resin particle that itself and diameter is bigger mixes, wherein the diameter of resin is 100～4000 μ m, resin accounts for cumulative volume about 5～30%.It is the capping of opening container top band filter screen behind the mix homogeneously in 3: 1～9: 1 according to volume ratio that length is had hepatocellular microcarrier and resin, and this mixture of packing into is built top closure then.
The advantage of cell reactor preparation method of the present invention is, used tank capacity carries out the cell loading operation, avoided in the hollow fiber bioreactor problem of uneven distribution behind the narrow and small regulating liver-QI cell device of the doughnut external space.Hepatocyte grows to desired density and growth conditions and is filled in the bioreactor when good again on microcarrier, having solved hepatocyte, directly to cultivate in bioreactor hepatic cell growth bad and be difficult to observe the problem of hepatic cell growth situation.
Another technical problem to be solved by this invention provides the renovation process of cell reactor, and its technical scheme is:
A kind of renovation process of above-mentioned cell reactor.This method comprises the cell reactor serum-free cell culture medium perfusion after using, after fully removing dialysis solution, its port of export is used the closed with covers that is sealed and matched with it, in container, add cell culture medium, arrival end is added a cover supporting with it lid, and lid unscrewed to the degree that allows air to see through, put into cell culture incubator and cultivate; Perhaps carry out perfusion by peristaltic pump with hepatocyte in the culture medium pair cell reactor and cultivate, according to the state and the patient treatment needs at interval of cytoactive, the time that regeneration is cultivated can be selected 1～7 day.Cell reactor after regeneration is cultivated can be recycled and reused for the preparation artificial liver.
The technical problem that the present invention will solve at last provides a kind of artificial liver support system, this artificial liver support system comprises apparatus for purifying blood and one or more above-mentioned cell reactor, described apparatus for purifying blood be any can the commercial apparatus for purifying blood that obtains, the dialysis solution port of export of described apparatus for purifying blood is connected with the fluid inlet end of described cell reactor, the liquid outlet end of cell reactor is connected with the dialysis solution arrival end of apparatus for purifying blood, constitute closed circuit, described a plurality of cell reactor arranged in series.
Artificial liver support system operation principle of the present invention is, in the blood circulation path, patient's arterial blood is flowed in the semipermeable membrane of doughnut by the dialysis cartridge that the blood inlet enters haemodialysis equipment, and the outer dialysis solution of semipermeable membrane flows in the opposite direction.Just entered the position of dialysis cartridge like this at blood, the blood middle high concentration noxious substance can enter dialysis solution by semipermeable membrane under the Concentraton gradient effect, along with noxious substance constantly enters dialysis solution, in the dialysis cartridge blood forward in the process of flowing toxin reduce gradually.And in the dialysis solution peripheral passage, liquid flow direction is opposite with direction of flow of blood, the port of export of blood inlet and dialysis solution is in the same side on dialysis cartridge, therefore the dialysis solution that contains toxin after the mass exchange flows out through outlet, enter cell reactor, after hepatocyte is detoxified in reactor, toxin reduces in the dialysis solution, the synthetic material of hepatocyte increases, therefore when dialysis solution by dialysis cartridge on dialysis solution enter the mouth when entering, the synthetic material of hepatocyte passes through semipermeable membrane under the Concentraton gradient effect, enter the blood of having removed toxin, this blood of having removed toxin and having increased the hepatocyte synthetic is got back in the human body under blood monitoring warning system monitoring by dialysis cartridge blood outlet outflow.
The advantage of artificial liver support system of the present invention is:
1. will load hepatocellular bioreactor and be connected on the dialysis solution peripheral passage of apparatus for purifying blood, connected mode is easy; And hepatocyte directly do not contact with blood, and biological safety is good.
2. adopt a plurality of bioreactor series connection, improve the detoxifcation efficient of artificial liver.
3. bioreactor is a cell reactor, is again a cell culture system, can repeatedly use by Regeneration Treatment, greatly reduces bioartificial liver's treatment cost.
Further specify the present invention below in conjunction with the drawings and specific embodiments, but should be pointed out that drawings and Examples example only of the present invention as an illustration, do not limit the scope of the invention.
Description of drawings:
Fig. 1 cell reactor sketch map of the present invention;
Fig. 2 artificial liver support system organigram of the present invention;
Fig. 3 bioreactor hepatocyte survival condition on the microcarrier of back of repeatedly regenerating, figure A-E is respectively regeneration and cultivates Gimsa dyeing photo after 0,1,3,5,7 time, and F is the hepatocyte survival percentage ratio that MTT measures.
The specific embodiment
The composition of embodiment 1 cell reactor
One, the composition of cell reactor
As shown in Figure 1, cell reactor of the present invention comprises a container 10, these container 10 bottoms are provided with liquid outlet 8, container top is provided with capping 3, capping 3 is provided with liquid inlet 2, be respectively equipped with one deck 100～400 purpose filter screens 7,4 in container bottom and the container top capping, liquid outlet 8 and inlet 2 places are provided with the lid 9,1 that is sealed and matched with it, hold the long mixture that hepatocellular microcarrier 6 and resin 5 are arranged between the two-layer filter screen of internal tank.
Load the long mixture that hepatocellular microcarrier and resin are arranged in the cell reactor of the present invention, wherein said hepatocyte is animal or human's primary hepatocyte, the human or animal's hepatic cell line or the tumor cell of liver of immortalization, described resin particle diameter is 100～4000 μ m, resin accounts for and loads 5～30% of cumulative volume, and it is 3: 1～9: 1 that described length has the volume ratio of hepatocellular microcarrier and mixed with resin.
In the composition of cell reactor of the present invention, above-mentioned resin particle diameter is preferably 300～1000 μ m, and preferably, resin accounts for and loads 10～20% of cumulative volume.
The preparation method of embodiment 2 cell reactors
Get Cytopore 2 microcarriers (available from U.S. Pharmacia company), the pretreatment before using is carried out according to manufacturer's explanation.Bioartificial liver's cell will be applicable to, for example the human liver cell of immortalization system is suspended among the serum-free medium Keratinocyte-SFM (American I nvitrogen company), wherein adds rEGF (final concentration 0.1ng/ml) and BPE (bovine pituitary extract) 20～30 μ g/ml).According to human liver cell is 5 * 10
7Cell: 1g microcarrier: the mixed of 500ml culture medium, under the situation of shaking, cultivate, cell is attached on the microcarrier.Changed liquid once every 2 days.Hepatocyte form on the microcarrier and quantity are observed by Gimsa dyeing.Treat that cell is grown when closely converging (80～100%) on the microcarrier, its diameter 800 μ m resin particles (available from Zhuhai Lizhu Medical Biomaterials Co., Ltd) with sterilization mixed that the volume ratio of microcarrier and resin particle is about 9: 1.Get the tank container that aseptic the present invention is used for the preparation feedback device then, use the closed with covers bottom opening, open the container top capping, this mixture of packing into is prepared into stand-by cell reactor.
Embodiment 3: the renovation process of cell reactor
After seance is finished, the cell reactor of biological liver cyclic part is pulled down, the closed bottom outlet, pour into serum-free cell culture medium from its top arrival end, open outlet at bottom after waiting to fill, liquid is flowed out from outlet at bottom under action of gravity, repeat 3～5 times with abundant removal dialysis solution.Treat then liquid flow out fully after with port of export closed with covers, in cell reactor, add cell culture medium, arrival end is added a cover lid, and lid is unscrewed to allowing air to see through, and puts into cell culture incubator and cultivates; Perhaps after cleaning dialysis solution fully, carry out perfusion by peristaltic pump with hepatocyte in the culture medium pair cell reactor and cultivate.Regeneration was cultivated after 12～24 hours, and the microcarrier that takes a morsel uses Gimsa dyeing observation of cell survival condition.Use the percentage ratio that MTT analyzes survivaling cell, recover then to abandon culture medium, with being used to prepare artificial liver once more after the dialysis solution washing as cell viability.So repeatedly, each cell reactor is repeatedly reusable.As shown in Figure 3, after regeneration is used 1,3,5,7 time on the microcarrier cell survival situation do not have obvious decline.
Embodiment 4: the composition of hybrid artificial liver support system
The artificial rami hepatici of the present invention system of holding comprises apparatus for purifying blood 13 and one or more embodiment 1 described cell reactor 22.Described apparatus for purifying blood is sophisticated instrument in the prior art; it is structure and operation principle for artificial liver support system of the present invention more clearly is described that present embodiment provides a kind of apparatus for purifying blood of structure, should not be construed as limiting the scope of the invention.
As shown in Figure 1, 2, apparatus for purifying blood 13 comprises blood monitoring warning system 14, dialysis cartridge and dialysis solution feed system 21, and blood access 11 that is connected with patient's tremulous pulse and the blood access 12 that is connected with patient's vein.Wherein dialysis cartridge is performance blood purification and antidotal main place, and it comprises dialysis solution peripheral passage 18, dialysis solution outlet 19, dialysis solution inlet 20 in dialysis cartridge inner blood peripheral passage 17, blood inlet 15, blood outlet 16 and the dialysis cartridge.Blood monitoring warning system 14 is connected in the outer blood circulation path of dialysis cartridge, the control blood circulation state; Dialysis solution feed system 21 is connected in the outer dialysis solution peripheral passage of dialysis cartridge, control dialysis solution recurrent state; Described cell reactor 22 is embodiment 1 a described structure.The dialysis solution port of export 19 of described apparatus for purifying blood 13 is connected with the fluid inlet end 2 of cell reactor 22, the liquid outlet end 8 of cell reactor 22 is connected with the dialysis solution arrival end 20 of apparatus for purifying blood 13, constitute closed circuit, described a plurality of cell reactor 22 arranged in series, its scale and quantity can be amplified according to the needs of practical application or dwindle.
In artificial liver support system of the present invention, comprise two peripheral passages: blood circulation path and dialysis solution peripheral passage.The blood circulation path be the blood lead body of patient or laboratory animal outer after, enter the blood monitoring warning system 14 of apparatus for purifying blood through blood path 11, enter dialysis cartridge inner blood peripheral passage 17 through dialysis cartridge blood inlet 15 then, flow out through dialysis cartridge blood outlet 16, at last under blood monitoring warning system 14 monitoring in blood path 12 is got back to patient's body.The dialysis solution peripheral passage is by 21 controls of dialysis solution feed system, dialysis solution dialysis solution inlet 20 on dialysis cartridge enters dialysis solution peripheral passage 18 in the post, dialysis solution outlet 19 flows out into cell reactor 22 on dialysis cartridge, cell reactor 22 effusive dialysis solution enter dialysis cartridge by inlet 20, so constitute the dialysis solution circulation.
Be separated with semipermeable membrane between circulation of dialysis cartridge inner blood and dialysis solution circulation, liquid flow direction is opposite with direction of flow of blood in the blood circulation path 17 in the peripheral passage 18 of dialysis solution, and blood and dialysis solution at this mass exchange take place.The result is that blood flow toxin concentration in the process of dialysis cartridge reduces gradually, and the synthetic biological substance of hepatocyte increases gradually, has therefore been detoxified by the hepatocyte in the cell reactor indirectly when blood flows out dialysis cartridge.
Embodiment 5: hybrid artificial liver support system is to the function of detoxification of liver failure patient blood plasma
For detoxification ability and the increase and decrease cell reactor quantity of estimating the novel recyclability hybrid artificial liver support system of the present invention influences function of detoxification, we utilize clinical liver failure patient's blood plasma that artificial liver support system of the present invention is estimated.At first according to embodiment 2 preparation artificial liver support system, liver failure patient's blood plasma with the 4L anticoagulant heparin is connected on the apparatus for purifying blood by blood access 11 then, start blood monitoring warning system with control blood circulation state, start the circulation of the dialysis solution that has connected cell reactor simultaneously.Carried out artificial liver treatment continuously 6 hours, bilirubin, urea concentration after measuring original bilirubin, urea concentration in the blood plasma then and having passed through apparatus of the present invention of different series connection quantity.The result is as shown in table 1, after artificial liver support system detoxifcation of the present invention, and blood plasma mesobilirubin lowering of concentration, urea concentration raises, and cell reactor of every increase, and effect also obviously increases.
In addition, we have also tested the effect of the cell reactor that regeneration uses, and the renovation process of describing according to embodiment 3 after each detoxifcation is finished to blood plasma, with the cell reactor Regeneration Treatment, and is used for antidotal therapy with a blood plasma.The result is as shown in table 2, and the cell reactor detoxifying effect that regeneration is used is good, and regeneration still has the better solutions toxic action after using 7 times.
Table 1: cell reactor quantity is held the influence of system's blood plasma Detoxication to the artificial rami hepatici of the present invention
Table 2: the cell reactor regeneration times is held system's blood plasma antidotal influence to the artificial rami hepatici of the present invention
Obviously, the above embodiment of the present invention only is for example of the present invention clearly is described, and is not to be qualification to embodiments of the present invention.For those of ordinary skill in the field, can also make other changes in different forms on the basis of the above description.Here can't give exhaustive to all embodiments.Everyly belong to the row that conspicuous variation that technical scheme of the present invention extends out or change still are in protection scope of the present invention.
Claims (10)
1. cell reactor, it is characterized in that, described cell reactor comprises a container, this container bottom is provided with liquid outlet, container top is provided with capping, and capping is provided with the liquid inlet, is respectively equipped with one deck 100～400 purpose filter screens in container bottom and the container top capping, liquid outlet and porch are provided with the lid that is sealed and matched with it, hold the long mixture that hepatocellular microcarrier and resin are arranged between the two-layer filter screen of internal tank.
2. cell reactor according to claim 1 is characterized in that, described hepatocyte is animal or human's primary hepatocyte, the human or animal's hepatic cell line or the tumor cell of liver of immortalization.
3. cell reactor according to claim 1 and 2 is characterized in that, described resin particle diameter is 100～4000 μ m, and resin accounts for and loads 5～30% of cumulative volume.
4. cell reactor according to claim 3 is characterized in that, it is 3: 1～9: 1 that described length has the volume ratio of hepatocellular microcarrier and mixed with resin.
5. the preparation method of cell reactor as claimed in claim 1, it is characterized in that, this method comprises hepatocyte is seeded on the microcarrier, treat hepatocyte grow on the microcarrier 80～100% converge after, there are hepatocellular microcarrier and mixed with resin to pack in the container length, are built into described cell reactor.
6. the preparation method of cell reactor according to claim 5 is characterized in that, the described method that hepatocyte is seeded on the microcarrier comprises: according to hepatic cell line 1 * 10
7～1 * 10
8The mixed of cell: 1～2g microcarrier: 500ml～1L culture medium is cultivated under the situation of shaking, and cell is attached on the microcarrier.
7. the preparation method of cell reactor according to claim 5 is characterized in that, it is 3: 1～9: 1 that described length has the volume ratio of hepatocellular microcarrier and mixed with resin.
8. the renovation process of cell reactor as claimed in claim 1, it is characterized in that, this method comprises the cell reactor after using, use the serum-free cell culture medium perfusion, fully remove dialysis solution after, with the liquid outlet end closed with covers of cell reactor, add cell culture medium, arrival end is added a cover supporting with it lid, and lid is unscrewed to the degree that allows air to see through, and puts into the cell culture incubator cultivation of regenerating.
9. the renovation process of cell reactor as claimed in claim 1, it is characterized in that this method comprises the cell reactor after using, uses the serum-free cell culture medium perfusion, after fully removing dialysis solution, carry out perfusion regeneration by peristaltic pump with hepatocyte in the culture medium pair cell reactor and cultivate.
10. artificial liver support system, comprise apparatus for purifying blood, it is characterized in that, also comprise the described cell reactor of one or more claim 1, the dialysis solution port of export of described apparatus for purifying blood is connected with the fluid inlet end of cell reactor, the liquid outlet end of cell reactor is connected with the dialysis solution arrival end of apparatus for purifying blood, constitutes closed circuit, described a plurality of cell reactor arranged in series.
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| CN101966352A (en) * | 2010-09-10 | 2011-02-09 | 浙江大学 | Bioartificial liver support system and combined type liquid storage tank thereof |
| CN102146356A (en) * | 2010-06-24 | 2011-08-10 | 辽宁成大生物股份有限公司 | Method for regenerating microcarrier |
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| US5270192A (en) * | 1991-02-07 | 1993-12-14 | Monsanto Company | Biological artificial liver |
| KR970706053A (en) * | 1992-09-11 | 1997-11-03 | 더블유. 제랄드 뉴민 | ARTIFICIAL LIVER APPARATUS AND METHOD |
| EP1063289A1 (en) * | 1998-03-03 | 2000-12-27 | JMS Co., Ltd. | Liver cell clones for artifical liver and extracorporeal liver assist device |
| CN201171818Y (en) * | 2007-10-31 | 2008-12-31 | 佛山市第一人民医院 | Protein circulation absorption filtration system |
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| CN102146356A (en) * | 2010-06-24 | 2011-08-10 | 辽宁成大生物股份有限公司 | Method for regenerating microcarrier |
| CN101966352A (en) * | 2010-09-10 | 2011-02-09 | 浙江大学 | Bioartificial liver support system and combined type liquid storage tank thereof |
| CN101966352B (en) * | 2010-09-10 | 2013-03-13 | 浙江大学 | Bioartificial liver support system and combined type liquid storage tank thereof |
| CN102199538A (en) * | 2011-03-14 | 2011-09-28 | 胡威 | Non-stirred bioreactor |
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| CN106620916A (en) * | 2017-01-23 | 2017-05-10 | 上海赛立维生物科技有限公司 | Artificial liver reactor as well as preparation method and applications thereof |
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| CN111704998A (en) * | 2020-07-14 | 2020-09-25 | 兰州大学第一医院 | Vertical plate constant-flow type biological artificial liver reactor |
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