200 results about "Perfusion Culture" patented technology

A three-dimensional (3-D) culture “Petri-dish” for research in regenerative medicine, biotechnology and clinical translation is described. This 3-D perfusion culture dish is to advance in vitro culture tools from static 2-D to dynamic 3-D perfusion culture. Interwoven hollow fiber capillary membranes divide the “Petri-dish” culture space into a controllable 3-D pattern of different compartments, serving the functions of the organ's larger vasculature. These physically active scaffolds, which can be suitable for cell adhesion or cell aggregate immobilization, offer a supply of cells with high-performance mass exchange including gas supply and under perfusion conditions. In contrast to static and discontinuous medium supply, a dynamic culture can be achieved with continuous or alternating medium supply and integral oxygenation. They provide a more physiologic supply in the cell macro environment, including homeostasis of oxygen, pH, nutrition, soluble factors, and gradients of metabolites for the cells. Also, medium perfusion can be achieved. Consequently the invention was made for cultures at tissue density, especially stem cells and support cells, which strive to create their own stem cell niche.

The invention discloses a system for perfusion culture of cells which comprises: a bioreactor; a filter unit with a retentate inlet end, a retentate outlet end and a permeate outlet port; a reciprocating pump fluidically connected to the retentate inlet end, with the retentate inlet end fluidically connected to the bioreactor via an inlet check valve arranged to allow flow in the direction from the bioreactor to the retentate inlet end and to block flow in the reverse direction, and where the retentate outlet end is fluidically connected to the bioreactor via an outlet check valve arranged to allow flow in the direction from the retentate outlet end to the bioreactor and to block flow in the reverse direction; and where the inlet and outlet check valves are each fluidically connected to a tubing branch point, which is further connected to the bioreactor via a length of tubing. In an alternative embodiment, each of said inlet and outlet check valves is fluidically connected to the bioreactor via separate lengths of tubing. The invention further discloses a pre-sterilized system for perfusion culture of cells which comprises: a bioreactor; a reciprocating pump and a filter unit fluidically connected to the reciprocating pump and fluidically connected to the bioreactor via at least one aseptic connector.

The invention relates to a perfusion bioreactor system which comprises a circulating perfusion system, a constant temperature system, a gas exchange system and an on-line monitoring system, wherein the circulating perfusion system comprises a culture chamber, a liquid storage bottle and a peristaltic pump which are sequentially connected through a pipeline to form a perfusion culture loop; the constant temperature system comprises a constant temperature water bath device of which the inside is provided with a circuitous circulating pipeline, and is applicable to an electric heating and refrigerating control part; the gas exchange system is composed of a gas filter, a gas flowmeter, a high-pressure gas bottle, a pressure relief valve and a connecting pipeline; and the on-line monitoring system is composed of a monitoring probe, a monitor terminal, monitoring software, a data conversion chip and a mass transfer regulator. The perfusion bioreactor system provided by the invention can maximally simulate the in-vivo microenvironment in the tissue or organ in-vitro construction process, ensures sufficient substance exchange and normal physiological functions of seed cells in the biological material, enhance the yield of the seed cell / biological stent material, shorten the culture period and lower the culture cost.

The invention discloses a high-yield reactor, and a production method and an application thereof. The high-yield reactor comprises a bioreactor and an ATF perfusion apparatus connected with the bioreactor. The protein production method using the high-yield reactor comprises the following steps: 1, connecting the ATF perfusion apparatus with the bioreactor, and completing an offline disinfection or online disinfection program; 2, inoculating seed cells to the bioreactor to a work volume according to the density, and sampling and detecting the number and biochemical indexes of the cells each 24h; 3, starting the ATF apparatus program for perfusion culture when the density of living cells increases to a certain density; and 4, collecting a culture solution in the bioreactor or filtered through hollow fibers in the ATF perfusion apparatus, and purifying to obtain required protein products. The high-yield reactor has the advantages of great shortening of the technological exploitation time of monoclonal antibody or fusion protein expression, production cost reduction, benefiting for the project registering and reporting and the project period shortening, and acceleration of the accomplishment application speeds of the biopharmacy industry.

The invention relates to a bioreactor for three-dimensional tissue cell perfusion culture. A reaction chamber, a liquid storage tank and a peristaltic pump are sequentially connected through transfusion pipes, wherein the reaction chamber comprises a box cover and a box body comprising lateral walls, a bottom wall, a cut-off device, an overflow insert plate, screen frames, a static pressure channel, a sluice channel and a draining channel; the cut-off device is a gate or a weir flow dam, and a plurality of uniformly arranged water holes are formed in the same plane along the longitudinal section of the cut-off device; an overflowing opening matched with the draining channel is formed in the lateral wall of the static pressure channel; and the screen frames are fixed on two lateral walls of the box body. The invention also relates to a bioreactor with a laminated structure, which can be used for solving the problem of cell death easily caused by non-uniform flow field and non-uniform nutritional transfer in the existing perfusion culture, is suitable for three-dimensional culture of tissue cells, can be used for improving the quality of tissue engineering products, and has the characteristics of good expansibility, strong universality and wide application range.

The invention discloses a technology for producing a recombinant antibody protein through the high density perfusion culture of hybridoma cells. The technology which aims at the metabolic characteristic of the hybridoma cells screens a specific culture component for the perfusion culture, and allows metabolic substrates to be fully supplied in the culture process and the concentration of metabolic byproducts to be simultaneously and effectively reduced, so objects of high efficiency and high yield is reached. In the process control aspect, a specific perfusion rate CSPR perfusion culture calculating model is established to carry out real time calculation on culture parameters, and concentrated medium subsection fed batch is adopted to control the perfusion rate, so important substrates ofglucose and glutamine are supplied and the influence of the byproducts to culture is minimized. The technology which is suitable for the preparation of the production antibody protein through the large scale culture of the hybridoma cells allows a large amount of protein antibodies to be provided in a short time, has the advantages of small scale, low investment, realization of the high efficiency output in a short time, low batch difference, high repeatability, and realization of the production of antibody medicines.

The present invention is related to a recovery and purification method of (a) biological(s) of interest produced by cells retained in a bioreactor under appropriate conditions and by appropriate means for producing the biological(s) inside the bioreactor having a volume higher than 1,5 l culture, and cultivated at a high cell density, preferably at a cell density higher than 10x10<6 >cells / ml, said bioreactor being submitted to an acoustic sonoperfusion allowing the recovery of a sonoperfused medium from said bioreactor, characterised in that it is submitted thereafter directly to an Expanded Bed Specific Adsorption for the direct recovery and uninterrupted purification of said biological(s). The present invention is also related to a process unit for recovering and purifying said biological(s) of interest.

The invention discloses a vascular tissue engineering reactor with rotation function. The reactor overcomes the disadvantages that the common arterial tissue engineering reactors can not simultaneously supply the vascular tissue with the culture conditions of intravascular perfusion and vascular rotation, and the mass transfer performance is poor. The vascular tissue engineering reactor comprises a liquid storage bottle, a liquid driving device, a vascular tissue culture cavity, a linear motor, a step motor, a vascular tissue transmission device, and the like, realizes the vascular tissue rotation in the course of intravascular perfusion culture, and has good mass transfer performance.

The invention relates to the field of tissue engineering, and in particular relates to a three-dimensional perfusion culture system and a tissue organ formed through 3D printing. The three-dimensional perfusion culture system comprises the following parts which are connected in series: a culture liquid supply part, a combined power source, a three-dimensional culture device and an oxygen / carbon dioxide circular supply part, wherein the culture liquid supply part is used or storing a culture solution for the three-dimensional perfusion culture system; the combined power source can simulate a heart supply system in the human body; the three-dimensional culture device enables the tissue organ to be cultured in a multi-freedom-degree movement manner; the oxygen / carbon dioxide circular supply part is used for simulating the lung respiratory system in the human body. With the three-dimensional perfusion culture system provided by the invention, a heart and lung simulation environment can be provided for the three-dimensional tissue organ formed through biological 3D printing and containing the tissue cells, and thus a microenvironment similar to the inside of the body is provided for the growth of the tissue organ. The invention further provides the tissue organ formed through 3D printing and cultured by the three-dimensional perfusion culture system.

The invention discloses a method for producing PRRS (Porcine Reproductive and Respiratory Syndrome) viruses through culturing Marc-145 cells by applying a torrent-perfusion bioreactor. The method comprises the following steps of: (1) selecting Marc-145 cells as cells for culturing seedlings; (2) subculturing the Marc-145 cells; (3) reproducing virus seeds for production; (4) perfusing and culturing the Marc-145 cells on a paper carrier in the torrent-perfusion bioreactor; (5) producing the PRRS viruses in the torrent-perfusion bioreactor; and (6) treating the obtained virus antigen solution. The invention can greatly decrease the production cost, improve the input-output ratio by more than 10 times and expand the production scale easily and rapidly, has short production preparation cycle and high degree of automation, occupies less area, causes less environmental pollution which is easy to treat, consumes less labor, is easy to realize balanced and stable virus quality and can obviously improve the yield and the quality of the viruses.

A rotatable perfused time varying electromagnetic force bioreactor with a rotatable perfusable culture chamber and a time varying electromagnetic force source operatively connected to the rotatable perfusable culture chamber. In use, the rotatable perfused time varying electromagnetic force bioreactor supplies a time varying electromagnetic force to the rotatable perfusable culture chamber of the rotatable perfused time varying electromagnetic force bioreactor to expand cells contained therein.

The invention discloses a modified AKTA pure chromatographic apparatus. On the basis of the structure of original standard AKTA pure, three multifunctional valves, as well as a second column position valve or a Loop ring valve, are added, so that two flow paths, which work simultaneously, are formed by three columns; one flow path is used for continuously loading a sample, while the other one flow path is used for pretreatment before and post-treatment after sample loading, so that continuous purification of a perfusion culture material solution is achieved. The modified apparatus can achieve low-cost, multi-cycle and automated purification of fed-batch culture solutions, avoids accumulation and degradation of the sample, can maintain the stability of the sample, is free of an extra large-volume buffer container, and is high in purification efficiency.

Disclosed is a method for obtaining differentiated cells and / or differentiated cell products, including the steps of: introducing undifferentiated cells into a perfused organ or living tissue; subjecting the undifferentiated cells introduced to perfusion culture together with the organ or living tissue so as to allow the undifferentiated cells to differentiate, thereby obtaining differentiated cells and / or differentiated cell products; collecting a perfusion culture solution that contains the resulting differentiated cells and / or differentiated cell products; and obtaining the differentiated cells and / or differentiated cell products contained in the collected perfusion culture solution.

The invention discloses a continuous pouring culture device of plant cell stirring typed biological reactor in the cell engineering technical domain, which comprises the following parts: one-grade sediment device, two-grade sediment device, three-grade sediment device, liquor storage bottle, sampling device, on-line monitor system, gas supplying device, sediment device in connection with the reacting tank on the magnetic stirrer, wherein the waste liquor bottle connects the sediment device. The invention is a group of three-gravity inner-outer cylinder biological reactor of sediment cell interception device to provide reference to culture large scale of plant cell, which improves the separating efficiency of the cell and culture liquid of reactor to reduce the probability of pollution.

A method for manufacturing a multilayered cell sheet characterized in fabricating a vascular bed that constructs a vascular network extending to the surface from a channel for perfusing a medium, the channel being embedded in a gel; and layering a cell sheet onto the vascular bed to construct a vascular network in the cell sheet. This manufacturing method makes it possible to construct a vascular network in the cell sheet and to fabricate a thick multilayered cell sheet in a simple manner by layering cell sheets. Such a thick multilayered cell sheet is useful as an in-vivo tissue substitute in regenerative medicine involving a variety of tissues.

The invention discloses an integrated dynamic inoculation culture method for an osteogenous seed cell and a matched perfusion type bioreactor thereof. A peristaltic pump, a perfusion chamber, a liquid storage bottle, a channel converter and a connection tube constitute a double loop structure for perfusion inoculation and perfusion culture. When the double loop structure is used, two loops are switched by regulating the channel converter, thereby achieving the effect of integrated dynamic inoculation and dynamic culture. The dynamic inoculation and dynamic culture of the cell are carried out through the perfusion type bioreactor, thereby enhancing the inoculation efficiency and uniformity of the cell, enlarging the range and efficiency of supplying nutrition and oxygen and simultaneously providing the shearing strength stimulus to the osteogenous cell so that the inoculation and culture of the cell are integrally carried out in the same environment, thus omitting a conversion link. Compared with the existing cell inoculation and culture technology, the invention has the advantage that a proper environment is provided for adhesion, distribution and differentiation of the cell and tissue construction, thereby obviously enhancing the effect of tissue engineering bone construction.

The present invention provides an incubator with a clean bench function to have both function of opened and closed system and a proposal how to use, allowing cells or others to be “closable and openable” manipulated and cultured, as well as an incubator, a globe box, a clean bench. This is implemented in this system has a specific structure and a characteristic component, e.g., a structure classified two units by the inside temperature, an operation cover and a circular fixture, a channel from / to inside or outside etc. to allow such as used, a non-opening operation and supply and observation from outside, or a moving of perfusion culture device and medium storing bags kept on connecting tubes or others. An incubator 100 with a clean bench function mainly includes a chamber 110, 210, 220, 230, a temperature control unit 160, 213 and a gas concentration control and supply unit 140, 240, as well as a CO2 cylinder 170, a N2 cylinder 180, and a N2 gas generator 190. The chamber 210 has an ability of setting on low temperature. The chamber 110 has the shape of a rectangular parallelepiped and includes an opening 111 in the front, and a sensor 161. The opening 111 attached on a sealing door 117 and an inner door 118, and their intervening space with elastic materials 120-122 and channels 126-128 in a peripheral portion 112 around the opening 111. For the closable operation inside the chamber 110 from the outside, an operation cover 136 and a circular fixture 300, a membrane with / without slit 134 and the fixer 134a, b is attached on / off the fixing hole 119 in the inner door 118, and a first hook 114 to hang of the operation cover is attached to each of two inner side surfaces 113 of the chamber 110. A UV lamp 116 adapted to emit UV rays for sterilization is attached on upper site of inner back surface 115 in the chamber 110. The N2 gas supplied in large quantity into the chamber from N2 cylinder 180 that positive pressure is maintained in the chamber 110 relative to outside pressure as used the clean bench function. In particular, this present invention is possible to mostly three new matters as follows: First, the exchange and regulate with gas composition and temperature in this system is both of them different constituent at the same time. Second, this system is an ability to operate under the condition of very low temperature (from 4 degree Celsius) and low O2 concentration (from 1%), too. Third, this system is an ability applied to variety of opened and closed operation etc. under the keeping with condition.

The invention relates to a perfusion operation method for large-scale culture of mammal cells, which is suitable for culturing the mammal cells with recombinant protein, and more suitable for culturing the mammal cells capable of producing TNFR-Fc fusing protein. The method has the following characteristics: firstly, engineering cells are amplified gradually in the generation and amplification process; secondly, the engineering cells are cultured in a of fermentation tank of 5L; and thirdly, during the later stage of the large-scale culture, the expression amount of the protein is improved byreducing the culturing temperature, lowering the pH value, reducing the rotation speed and adjusting the filling speed rate. According to the process, the production period is prolonged, the cell density is increased, the cell viability is improved, and the expression amount of the protein is improved greatly.

The invention discloses an extraction and perfusion culture system for MSCs (Mesenchymal Stem Cells), belongs to a field of medical instruments, and more specifically, provides a set of continuous and completed system design for obtaining high-purity stem cell products from a placenta organ. The device provided by the invention comprises an extraction device and a perfusion culture system, and the extraction device and the perfusion culture system are connected hermetically and aseptically, wherein the extraction device comprises a placenta cutting device, a reaction tank, a filter device, a filtrate collection device and a density gradient centrifugation device; and the perfusion culture system comprises a culture solution storer, a monitoring reactor, a cell culture bottle, a liquid waste bottle, a bottle for obtaining liquid, a biochemical criterion detection head group, an online monitoring system, a speed control fluid propeller and a pipeline device capable of controlling speed. The extraction and perfusion culture system for the MSCs (Mesenchymal Stem Cells) has benefits that the cutting efficiency is high; the operation is convenient and effective; the production efficiency can be greatly improved; the preparation technology is simple; and the quality control is simple and convenient, so as to efficiently obtain uniform and good stem cell products within a short time.

The invention provides a method for producing a recombinant adenovirus with serum-free suspension cells. The method comprises the following steps: (1) cell expansion; (2) inoculation into a bioreactor, culture solution perfusion culture and cell re-expansion: inoculating cells harvested from spinner bottles into the bioreactor to be cultured, wherein the pH value and the dissolved oxygen (DO) value of the culture solution are automatically controlled by a controller of the bioreactor via the pH value and DO probes, the pH value is controlled between 6.5 and 8.2, DO is controlled between 15% and 75%, the temperature of the culture solution is automatically controlled between 36 DEG C and 37 DEG C by an electric heater in a platform of the bioreactor, the swing speed is controlled between 15rpm and 20rpm, and the swing angle is controlled between 8 degrees and 15 degrees; (3) cell dilution and cell transfer to the bioreactor with big volume; (4) recombinant adenovirus infection and expansion. The method can achieve large-scale industrialization and can be used for preparing recombinant adenovirus products safely, economically and effectively according to GMP (good manufacture practice).

The invention relates to a double-loop oscillation perfusion-type biological reaction system which is applied to co-culture of seed cells and biomaterials. The biological reaction system comprises a support perfusion culture device, a liquid storage bottle, a recovery bottle and peristaltic pumps, wherein the support perfusion culture device is internally provided with a plurality of supports for co-culture of seed cells and biomaterials; the liquid storage bottle is used for conveying a culture solution in the bottle to the supports; the recovery bottle is used for receiving the culture solution flowing from the support perfusion culture device through a liquid conveying pipe; the flowing direction of the liquid storage bottle and the flowing direction of the recovery bottle are controlled through respective three-way pipes, so that the liquid storage bottle and the recovery bottle are switched between a perfusion liquid-changing channel and a perfusion culture channel; the peristaltic pump is respectively arranged in the perfusion liquid-changing channel and the perfusion culture channel and used for controlling the flow velocity of a culture solution passing through the liquid conveying pipe. According to the double-loop oscillation perfusion-type biological reaction system, the combination between an oscillation method and the perfusion method is achieved, the seed cells inside the biomaterials are subjected to sufficient substance interchange and stress stimulation, the production efficiency of engineered tissues is improved and the requirements of large-scale classed culture can be met.

The present invention is perfusion culture medicine method and apparatus for hematopoietic cell. The apparatus includes at least stirring bioreactor and perfusing culture cavity connected via pipeline to the stirring bioreactor. The perfusion culture process includes first static perfusion culture until reaching higher cell density, and subsequent stirring suspension culture to eliminate the cell density gradient and accelerate cell growth. Test result shows that the initial culture period has cloning in great amount of total cell as well as CFU-Mix, CFU-GM and CD34+ cell, and the subsequent stirring suspension culture stage has further cloning of total cell as well as CFU-Mix, CFU-GM and CD34+ cell. The said method has greater cloning than single stirring suspension culture.

The invention discloses a continuous filling culture gas-lift biological reactor system of medicinal plant tissue cell in the biological technical domain, which comprises the following parts: liquid storage bottle, reactor host, gravity sediment device, waste liquor bottle, tissue / organ continuous acquiring device, duplex sampling bottle, liquid pump, oxygen probe, pH value probe, on-line monitor system and computer, wherein the liquid storage bottle, liquid pump, reactor host, gravity sediment device and waste liquor bottle connect through silica gel pipe sequently; the oxygen probe and pH value connect computer through on-line monitor system. The invention can be applied for culturing kinds of different tissue, cell and organ to save cost, which avoids the damage of plant tissue cell for medicinal plant tissue, cell and organic suspension culture.

A cell depositing interception device of hematopoietic cell continuous perfusion cultivation bioreactor system, comprises an inclined tube-form depressor with tube-form cross-section, the upper portion of which is arranged with a feed inlet, a discharge port is arranged on the bottom of the depressor corresponding to the feed inlet position, the upper surface of the other end is provided with at discharge port and an exhaust port.

Methods of expanding T lymphocytes in a microfluidic device are provided. The methods can include introducing one or more T lymphocytes into a microfluidic device; contacting the one or more T lymphocytes with an activating agent; and perfusing culture medium through the microfluidic device for a period of time sufficient to allow the one or more T lymphocytes to undergo at least one round of mitotic cell division. The expansion can be non-specific or antigen-specific. T lymphocytes produced according to the disclosed methods are also provided, along with methods of treating cancer in a subject. The methods of treating cancer can include isolating T lymphocytes from a tissue sample obtained from the subject; expanding the isolated T lymphocytes in a microfluidic device; exporting the expanded T lymphocytes from the microfluidic device; and reintroducing the expanded T lymphocytes into the subject.

The invention discloses a high-flux dynamic culturing system for 3D cells, tissues and organoids. The high-flux dynamic culturing system comprises a machine shell body, culture solution mixing tanks,peristaltic pumps, a perfusion culturing chamber, a waste solution storing chamber and a pure air source, wherein the culture solution mixing tanks, the peristaltic pumps, the perfusion culturing chamber and the waste solution storing chamber are sequentially arranged in the machine shell body in the transporting direction of culture solutions, and the pure air source is arranged at the side partof the machine shell body and is communicated with the culture solution mixing tank. The system provided by the invention can be used for high-flux dynamic culturing of 3D cells, tissues and organoids; and through the peristaltic pumps, the air-enriched culture solutions can be transported into high-flux perfusion culturing boards at a perfusion culturing region, 3D dynamic culturing of three dimensional cultured cells, the tissues and the organoids can be automatically completed. The high-flux dynamic culturing system for 3D cells, tissues and organoids disclosed by the invention is safe andreliable in the whole course, efficient and convenient, the three dimensional culturing requirements of researchers in different fields can be met, and the high-flux dynamic culturing system conformsto the developing direction of biomaterials and cell biology and has high popularization and application value.

The invention provides a proliferation culture medium for 3D simulation culture of stem cells, the proliferation culture medium is free of serum addition, definite in composition and suitable for 3D simulation perfusion culture of stem cells, and comprises inorganic salt, amino acid, saccharides, lipids, nucleosides, vitamins, cell metabolites, cell proliferation factors and water, the osmotic pressure of a culture solution is 270-330mOsmol / kg.H2O, and the pH value of the culture solution is 7.2-7.4.

The invention relates to a proliferation culture medium for 3D simulation culture of stem cells, the proliferation culture medium is free of serum addition, clear in composition and suitable for 3D simulation perfusion culture of stem cells, and comprises inorganic salt, amino acid, saccharides, lipids, nucleosides, vitamins, cell metabolites, cell proliferation factors and water, the osmotic pressure of a culture solution is 270-330mOsmol / kg.H2O, and the pH of the culture solution is 7.2-7.4.

The invention discloses one filteration- deposition combined perfusion system used for animal cell perfusion culture. It comprises filtering- deposition- perfusion column, peristaltic pump, automatic controller, tri- way pipeline and pipe clamp; filtering- deposition- perfusion column is connected with peristaltic pump through tri- way pipeline, the rotation of peristaltic pump is controlled by automatic controller, pipe clamp is installed on pipe between filtering- deposition- perfusion column and peristaltic pump. The invention is characterized by simple structure, low cost, high collecting efficiency, long life length and wide application on micro- carrier or cluster animal cell perfusion culture.

The invention discloses a recombinant human erythropoietin-CTP fusion protein production process. The process is characterized in that a microcarrier suspension perfusion culture technology or a serum-free suspension fed-batch culture technology is adopted as a recombinant engineering cell line culture manner. The invention also discloses a recombinant human erythropoietin-CTP fusion protein purifying process. Through optimization, a simple process flow is obtained, such that high-purity and high-activity recombinant human erythropoietin-CTP fusion protein is obtained. The invention also relates to the ticokinetics study of the recombinant human erythropoietin-CTP fusion protein in monkey bodies, and the application of the recombinant human erythropoietin-CTP fusion protein in treating rat renal anemia.