134 results about "Artificial liver" patented technology

Artificial liver devices and methods for using the devices to purify a biological fluid are disclosed. The methods include the use of living hepatocytes (23) which are either unattached or attached to inert carriers and suspended in a cell culture medium which circulates in the devices with the hepatocytes (23). Blood or plasma passes on one side (7″) of semi-permeable membranes, on the other side (7) of which is the cell culture medium and across which is a concentration and/or pressure gradient. Solutes diffusing across the membrane into the cell culture medium are metabolized by the hepatocytes (23) and/or captured by additional removal means (4). Those undesirable substances which do not diffuse out of the blood or plasma into the hepatocyte containing culture medium are captured by additional removal means (50).

Disclosed is a 3-D bioreactor to culture hepatic cells, comprising a culture cavity provided with a cell feeding port and a vent, an inflow cavity with a culture fluid inlet and an outflow cavity with a culture fluid outlet which are arranged next to the culture cavity. A plurality of hollow fibers are arranged inside the culture cavity, each with one end communicated with the inflow cavity and the other end communicated with the outflow cavity, forming a culture fluid flow passage which is communicated with the culture cavity through the tiny holes on the hollow fibers. The invention is characterized in that a 3-d bracket with a plurality of holes is arranged inside the culture cavity. The invention can provide a growth and culture condition that accords better with the physiological environment for the hepatic cells, which is good for culturing dense hepatic cells with high activity so as to ensure the therapeutic effect of a bio-artificial liver with the hepatic cells as the core.

A drug testing system using a liver-slice culture apparatus. The apparatus has a chamber with plasma and gas valves, animal liver slices being positioned securely in the chamber so as to maximize the surface area of liver slices exposed to the culture medium. Plasma is supplied to the chamber so that it rises to contact the liver slices, and is alternately removed from contacting the liver slices. Gas is supplied to the top of the chamber. The system also includes a reservoir for containing media entering and exiting the chamber. Methods are provided for assessing the toxicity of a drug or drug candidate.

The invention discloses a microcapsule floating fluid-bed biological reactor of artificial liver in the medically biological technical domain, which is characterized by the following: displaying hopper shape with small bottom radius and large top radius; setting cell screen 8 at upper neck and cell screen 7 at lower bottom; depositing cell filter 8 at liquid outlet 2; setting buffer cavity 12 between liquid inlet 1 and cell screen 7, and between cell filter 11 and cell screen 8 respectively; containing the microcapsule liver cell 9 between cell screen 8 and cell screen 7; making the liquid inlet connect the membrane oxygenator 14 to connect immune adsorber 15 to adsorb immune macromolecule. The invention makes the moving trail of flow impaction as adjustment parabola to satisfy the character of fluid mechanics, which avoids dead perfusion cavity and ineffective perfusion to develop treating effect of biologically artificial liver obviously.

The invention discloses a low-expression CYP7A1 hepatic cell and a constructing method thereof. A method for inhibiting the CYP7A1 expression level of the hepatic cell comprises the step of introducing an encoding gene of small-interference RNA which is used for inhibiting the CYP7A1 gene expression into the hepatic cell so as to inhibit the CYP7A1 expression level of the hepatic cell. A slow virus vector system can be used for introducing the encoding gene of small-interference RNA which is used for inhibiting the CYP7A1 gene expression into the hepatic cell. An experiment proves that the method of the invention can obviously regulate the expression level of the CYP7A1 gene of the hepatic cell in an mRNA level and a protein level downwards and can further effectively reduce the secretoryvolume of total bile acid. The Low-expression CYP7A1 hepatic cell and the constructing method thereof can be used for the fundamental research and the clinical application of biologic artificial liver supporting treatment and have a wide application prospect.

The present invention relates to a novel tissue culture system that provides for the long term culture of proliferating hepatocytes that retain hepatic function. Disclosed are methods and compositions for ex vivo culturing of hepatocytes and nonparenchymal cells on a matrix coated with a molecule that promotes cell adhesion, proliferation or survival, in the presence of growth factors, resulting in a long-term culture of proliferating hepatocytes that retain hepatic function. The co-culturing method results in the formation of matrix/hepatic cell clusters that may be mixed with a second structured or scaffold matrix that provides a three-dimensional structural support to form structures analogous to liver tissue counterparts. The hepatic cell culture system can be used to form bio-artificial livers through which a subjects blood is perfused. Alternatively, the novel hepatic cell culture system may be implanted into the body of a recipient host having a hepatic disorder. Such hepatic disorders, include, for example, cirrhosis of the liver, induced hepatitis, chronic hepatitis, primary sclerosing cholangitis and alpha1 antitrypsin deficiency.

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.

The invention provides preserving fluid of hepatic cells for a biological artificial liver and a preparation method thereof. The preserving fluid is a solution compounded by ultrapure water. The solution contains the following components within the concentration range: 15-25mmol/L of disodium hydrogen phosphate, 1-10mmol/L of sodium hydrogen phosphate dehydrate, 4-6mmol/L of potassium citrate monohydrate, 10-30mmol/L of sodium chloride, 5-10mmol/L of magnesium chloride hexahydrate, 3-10mmol/L of disodium adenosine triphosphate, 1-5mmol/L of reducing glutathione, 0.1-0.5mmol/L of alpha-lipoic acid, 100-150mmol/L of trehalose (C6H12O5), 200/0.510-50g/L of hydroxyethyl starch and 2-10mg/L of matrine. The preparation method of the preserving fluid comprises the following steps of: accurately weighing all components according to the concentration requirements of the components, wherein the alpha-lipoic acid is weighed in a dark place; completely dissolving the other components except the alpha-lipoic acid by using the right amount of ultrapure water; sufficiently dissolving the alpha-lipoic acid in the dark place; and adding the ultrapure water to full dose. The preserving fluid can well protect the cell activity of the hepatic cells for the biological artificial liver and the special functions of the hepatic cells at low temperature so as to satisfy the short-term low temperature preservation of a large-scale hepatic cell bank for the biological artificial liver and/or the hepatic cell protection in the long-distance transportation process.

The invention provides an artificial liver tissue and a preparation method thereof. The artificial liver tissue comprises a lower chip, a first microporous membrane, a middle chip, a second microporous membrane and an upper chip from the top to the bottom in sequence in a laminated manner. Three layers of cavities of a lower cavity, a middle cavity and an upper cavity are formed in the lower chip, middle chip and upper chip respectively. Three cavities are all grouted with culture solution containing growth factor, extracellular matrix containing hepatic parenchymal cells and suspension liquid containing endothelial cells. Adjacent cavities are separated by the microporous membrane. The microporous membrane can separate the matrix flow outside of the cell, and cause no influence to the cell migration. The culture solution and is fed through an inlet and an outlet of the upper cavity and lower cavity. Under the guidance of the growth factor, endothelial cells pass through the extracellular matrix of the middle cavity to form a lumen, and form the artificial liver tissue containing capillaries. The artificial liver tissue can simulate the human body liver tissue in a good way, and simulate the intrahepatic capillary functions of mass transfer, selective permeability, toxic reaction and the like.

The invention provides a kit used for obtaining a full liver stent, comprising perfusion liquid I: 0.1 to 10 percent (volume fraction) of Tris-HCl solution of non-ionic detergent; and perfusion liquid II: 0.1 to 10 percent (mass fraction) of Tris-HCl solution of ionic detergent. The invention also provides a method for obtaining the full liver stent by using the kit. The invention leads the substantive organ liver to be acellular to completely and simultaneously reserves the structure and large part of components of extracellular matrix, thus being used as materials of biological stents and being applied to research of biological artificial liver.

The invention discloses a dual-cavity liquid storage tank for a biological artificial liver support system. The dual-cavity liquid storage tank comprises an inner cavity and an outer cavity, wherein the top of the outer cavity is provided with a first liquid inlet and a first air pressure balance hole; the top of the inner cavity is provided with a second liquid inlet and a second air pressure balance hole; the bottom of the inner cavity is provided with a first liquid outlet; the side wall, which is close to the bottom, of the inner cavity is provided with a second liquid outlet; the second liquid outlet passes through the outer cavity by means of a pipelike passage, and directly leads to the outside of the dual-cavity liquid storage tank from the inner cavity; and the pipelike passage is provided with a plurality of outer cavity side holes on the partial side wall of the outer cavity. The dual-cavity liquid storage tank for the biological artificial liver support system realizes that severe hepatitis plasma flows through a bioreactor for multiple times to form a high-efficiency circulation path, thereby playing better therapeutic effect. The proportion of volume sizes of the inner cavity and the outer cavity of the dual-cavity liquid storage tank for the biological artificial liver support system can also be reset according to clinical concrete practical requirements, and is expanded or reduced so as to be applied to clinic treatment more reasonably.

The present invention provides a method that embryo of human embryonic stem cells (HES) is directionally induced and divided into liver cells. First, the biological property of high degree of self-regeneration multiplication of HES cell is made use of, the collagenase IV digestion is adopted for transfer of culture and augmentation, thereby obtaining a plurality of stem cells; second, the HES cell is inoculated into a cell utensil with lower adsorbability to form a mature imitated embryonic plant (EB); third, the mature cystic EB is digested with 0.25 tryptic enzyme to 0.02 percent EDTA to a single cell, and then is inoculated to a tissue culture dish which is packed with I-shaped collagen in advance, solution culture is induced under the condition of dexamethasone and human trypsin, and the differentiation towards the liver cell direction is observed and judged. The prevent invention provides the method that the HES is efficiently divided into liver cells, at the same time, the method makes the follow-up judging work more simply and more efficient, and lay a foundation for the stem cellular transplantation or biologic artificial liver curing the disease such as serious hepatitis, hepatic failure and so on.

The invention discloses an immortalized human liver cell line, a preparation method and an application thereof. The immortalized human liver cell line has the main function of primarily cultured humanliver cell. The preparation method comprises the following steps of: transferring SV40T antigen and human telomerase catalyzed subunit gene into primarily cultured separated human normal liver cell by slow virus carrier; and sieving in a cloning way to obtain the immortalized human liver cell line. The immortalized human liver cell line also can be extrinsically amplified in large scales in a microcarrier way. The immortalized human liver cell line can be used for preparing a biological artificial liver support system and preparing the medicament for curing the liver failure.

The invention discloses an artificial liver reactor. The artificial liver reactor comprises an input header pipe, an output header pipe, and one or more cell culture bottles connected in parallel or in series, wherein the cell culture bottles are connected between the input header pipe and the output header pipe, an input pipe and an output pipe are arranged at two sides of the bottom of each cell culture bottle, and a peristaltic pump is arranged on each input pipe; a separation net is arranged in each cell culture bottle, the part between the separation net and the bottom of each cell culture bottle is filled with a filler with an adsorption function, a flake-shaped carrier is loaded to each separation net, and hepatic cells attach to the flake-shaped carriers. For the artificial liver reactor, due to the arrangement of the peristaltic pumps and the flake-shaped carriers, the cell loading capacity, the effective exchange area and the metabolism exchange efficiency of the artificial liver reactor can be effectively improved. The invention further discloses a method for preparing the artificial liver reactor, and a circular pouring bioartificial liver support system comprising the artificial liver reactor.

The invention discloses a construction method of a three-dimensional liver microtissue unit for a bio-artificial liver support system. The construction method is characterized by comprising the steps that decellularized scaffold components of a pig liver are encapsulated by microcapsules prepared from sodium alginate-polycations, liver cells, vascular endothelial cells and fibroblasts are composited, and the functional three-dimensional liver microtissue unit is constructed through cell self-assembly and applied to the bio-artificial liver support system. According to the method, operation is easy, cost is low, the liver cells, the vascular endothelial cells and the fibroblasts are self-assembled into three-dimensional liver microtissue by simulating interaction between the cells and an extracellular matrix, interaction between the cells and soluble factors and interaction between the cells and the cells in a liver tissue microenvironment, the three-dimensional liver microtissue has good synthesis, secretion and detoxification functions, and therefore the three-dimensional liver microtissue can serve as the microunit to be applied to the bio-artificial liver support system.

The invention discloses a method for producing the packed liver project bracket. First according to the inner micro-structure of natural liver, processing the bionic design of liver project bracket blood vessel system and the liver hole; using the fast molding micro-complex technique to produce the silicon rubber mould of bionic liver project bracket; plating the biological material with better bioavailability and biodegradability on the silicon rubber mould; packing the liver project bracket mould after removing mould to realize the spatial structure of liver project bracket; and wetting, sopping, cooling and drying the liver project bracket to attain the spatial liver project bracket with high porous property. When in the external culture, the endothelial cell of blood vessel is planted on the blood vessel network system, and the liver cell is planted on the liver cell hole; then, with the degradation of spatial liver project bracket, the endothelial cell can form the blood vessel, and the liver cell can increase and reunite to grow into the liver organization, to supply the base of transforming the artificial liver into natural liver.

Disclosed is method to prepare biological artificial pig for liver and liver cell of human, which includes the two courses of pouring outside the body of separating liver cell and washing the liver cell, uses the four-stepped pouring method on the basis of the one with two steps, and designs the relative agent. The invention has increased the quality, living rate and purity of separating the liver cell, it is suitable for acquiring liver from pig, dog or grown-ups, or liver separation from liver of human embryo by legal artificial labor, and the liver cell prepared can be widely used in research and transplantation of liver cell, biological artificial liver and mixed artificial liver.

This invention relates to an artificial live bioreactor with nanofiber scaffold material as the culture medium for liver cells. A plasma inlet and an oxygen inlet are set at one end of the bioreactor hull, while a plasma outlet and an oxygen outlet are set at the other end. A nutrient solution inlet and an outlet are set at both ends of one side of the bioreactor hull, respectively. A culture medium composed of nanofiber scaffold and hollow fibers is set in the bioreactor hull. The nanofiber scaffold and the hollow fibers form coiled sandwich structure, with the nanofiber scaffold coiled into spiral drum and the hollow fibers encapsulated in the interlayer. The liver cell aggregates are adhered to the hollow fibers. An oxygen passage is set in the bioreactor to realize real-time oxygen supplement.

The invention relates to medical facilities, aiming at providing a hybrid artificial liver support system equipped with a nano fibre sheet stacked reactor. The system comprises a plasma separating part, a nonbiological liver circulating part and a biological liver circulating part which comprises a thermostatic gas bath incubator; an extracorporeal circulating pool, a plasma pump and dissolved oxygen which are connected sequeitically and form a circulating system; a temperature testing bottle; and the nano fibre sheet stacked reactor. The system is easily to be amplified so as to improve the defect that cells block a semipermeable membrane easily, the efficiency of material exchange is high and both oxygenation and immunity obstruction function are provided. The reactor has strong function, can be used for biological or hybrid artificial liver, and also used for the cultivation and inducing differentiation of stem cells. The reactor of the system has oxygenation function, thereby saving an oxygenator needing to be equipped independently in normal systems.

The invention discloses an artificial liver and kidney supporting system, which is characterized by the following: connecting output tube of human body blood with blood inlet of the filter; connecting the blood inlet of the filter to blood inlet of plasma separator; communicating the outlet of the plasma separator with inlet of venous pot; connecting input pipe of human body blood to the outlet of the venous pot; connecting film outer up interface of the filter to waste liquid bag through waste liquid pipe; cascading heater with isosmotic solution bag; dividing to two road; connecting one road to film outer lower interface through the second peristaltic pump; communicating the third peristaltic pump with the inlet of the venous pot; connecting the film outer lower interface of the plasma separator to carrier pipe of the separating matter; cascading the separating matter carrier pipe with infusion device through the forth peristaltic pump; communicating with the inlet of the venous pot. This invention increases survival rate of liver and renal failure patient.

The invention discloses a bioreactor for artificial liver, wherein the conventional hollow fiber inner core bioreactor is used as a basis, nonwoven fabrics are employed to enclose a plurality of hollow fiber inner cores, thus the hollow fiber inner cores can be attached to the nonwoven fabrics with the gaps sustained. The nonwoven fabric windings are arranged parallel in the cylindrical container, the nonwoven fabrics have strong affinity to hepatocyte, thus the hepatocyte is easy to be adhered to the nonwoven fabrics walls for exerting its biological functions.

The invention discloses a deflector-type micro-capsule suspension type fluidized bed bioreactor for an artificial liver. The deflector-type micro-capsule suspension type fluidized bed bioreactor for the artificial liver comprises an end cover part, a main body part and a base part, wherein the base part comprises an inlet buffer tank, a fixed turbine guide vane, and an inlet filter net arranged at the joint surface of the base part and a subject part; the main body part comprises a cylindrical containing cavity, a sampling and micro-capsule pouring lip and a valve; the sampling and micro-capsule pouring lip and the valve are arranged in a housing of the main body part; and the top cover part comprises an outlet buffer tank, and an outlet filter net arranged at the joint surface of the subject part and the top cover part. By adopting the sealed structure and a multi-opening design, the microencapsulated hepatocyte can be conveniently sampled and added; and meanwhile, the possibility of pollution in an operation process is reduced. The turbine guide vane of the base part can disperse large local flow speed at the inlet; micro-capsule damage is reduced; and the blood purification efficiency is improved. By adopting the deflector-type micro-capsule suspension type fluidized bed bioreactor for the artificial liver, the contact efficiency of the cultivated cell in the bioreactor and perfusate can be improved; meanwhile, the cyclic purification efficiency of the biological part in an artificial liver system is improved; and clinic application is facilitated.

The invention discloses a filling rack perfusion biological reactor of artificial liver in the medically biological technical domain, which is characterized by the following: the appearance is cylinder shaped container; the liquid outlets 1 and 2 are upper and lower ends of the cylinder container; the outlet 3 and 4 are distributed in the center of lateral wall symmetrically; the physical rack 6 has 14-16 biological racks as crossing arrangement to deposit separation area and support liver cell; the liquid buffer area 7 is set in the middle of the inner rack of the reactor; setting cell screen and cell screen brush at outlet; connecting the upstream of two outlets of biological reactor with membrane oxygenator 14; connecting the downstream of two outlets with immune adsorber 15; the biological rack of the liver cell is chitose rack or alginic acid/chitose and modified alginic acid/galactose chitose/heparin rack; possessing good biological compatibility; fitting for growing liver cell and developing the function; avoiding collapse of the biological rack of the liver cell.

The invention relates to a method for preparing a porcine hepatocyte and mesenchymal stem cell co-microencapsulated internal bio-artificial liver. The invention comprises the following steps: establishing an in-vitro culture amplification system for porcine mesenchymal stem cells, performing in-situ perfusion separation of porcine primary hepatocytes by a two-step collagenase method, and preparing alginate-poly-L-lysine microencapsulated hepatocytes and mesenchymal stem cells. The invention overcomes the defects that internal cells usually can not survive for a long time; the proliferation ability is quite limited; and the function of the injected hepatocytes for generating various stimulating factors for promoting liver regeneration is inhibited. The invention introduces mesenchymal stem cells into an internal bio-artificial liver system, fully simulates the in-vivo microenvironment of hepatocytes by heterogeneous intercellular interaction, effectively prolongs the survival time of hepatocytes, promotes the proliferation and differentiation of hepatocytes, maintains the specific biological functions of hepatocytes, and thus significantly improves the efficacy of the internal bio-artificial liver on treating hepatic failure.

The present invention relates to the field of extracorporeal liver perfusion and, more particularly, to the design of a chamber in which a biological component can be housed to form e.g. a bio-artificial liver (BAL). It also relates to a bio-artificial liver per se, it's components and methodological steps associated with its development and use. The chamber (10) for the biological component (100) of a bio-artificial liver (200) is configured to allow: • Proliferation of the biological component, in situ; • Cryopreservation of the biological component, in situ, and • Perfusion of the biological component, in situ.

The invention discloses a reciprocally turning perfusion bioreactor for biological artificial liver and a processing method thereof. The invention discloses a bioreactor, which comprises a cylindrical container, wherein the upper and lower ends of the circumferential side wall of the container are both provided with a liquid inlet, and the two end parts of the container both have a liquid outlet. Pressure buffering rings are arranged at the positions of the liquid inlets in the container respectively, and cell screens are arranged at the positions of the liquid outlets at the two ends parts in the container respectively. The invention also provides a reciprocally turning perfusion bioreactor for biological artificial liver, which has the bioreactor, and a processing method thereof. The bioreactor has the advantages that: liver cells cultured on a micro carrier are uniformly distributed in the entire bioreactor; and in a reciprocating motion, the bioreactor causes little damage to the liver cells, allows liver failure plasma to be contacted fully for effectively avoiding perfusion dead cavity and ineffective perfusion, can detoxify plasma with high efficiency, is simple in operation, low in cost and remarkable in effect, and allows the biological artificial liver to play a treatment effect to a maximum degree.

A bioartificial liver system has a separator for separating plasma and blood, a liver-slice culture apparatus, or bioreactor, to detoxify the plasma. The bioreactor has a chamber with plasma and gas inlets, at least two meshes mounted parallel one above the another, near the upper portion of the chamber forming at least two horizontal layers separated by a space. A plurality of liver slices positioned within the space, a supply of plasma is provided to the chamber so that it rises to contact the liver slices, and is alternatively removed from contacting the liver slices, and a supply of gas is provided to the top of the chamber. The system also includes a reservoir for containing plasma entering and exiting the chamber. Methods are provided for detoxifying plasma using the bioartificial liver system.