199 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 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 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.

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.

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.

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 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 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.

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.