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108 results about "Cell Maintenance" patented technology

Processes that promote, sustain, and preserve cell viability. (NCI)

Method for separating and extracting stem cells from placenta, umbilical cord or adipose tissue

The invention relates to a method for separating and extracting stem cells from a placenta, umbilical cord or adipose tissue, which comprises the following process steps: firstly mixing the placenta, umbilical cord or adipose tissue and a cell maintenance fluid according to the proportion by weight of 2.5-4:1, putting the mixture into a tissue crushing barrel, adding collagenase after crushing, uniformly mixing, hatching at the temperature of 37 DEG C, filtering, adding a precipitator, sucking a supernatant fluid after settling, centrifuging, removing the supernatant fluid, adding concentrated cells into a liquid of diatrizoate sodium-ficoll 400#, then centrifuging, collecting 10-15 ml of intermediate cell layer, washing with the cell maintenance fluid, counting the collected cells, and providing the cells for clinical use when the cell survival ratio is more than or equal to 95 percent. The invention not only realizes the separation and the extraction of all stem cells from the placenta, umbilical cord or adipose tissue, but also realizes industrialized production, and enables doctors to conveniently, safely and canonically obtain the adult stem cells in clinic and use the adult stem cells for treating the diseases of patients as using medicaments, thereby solving the bottleneck problem of difficult obtainment of adult stem cells in clinic and popularizing a cell treatment technology.
Owner:NINGXIA ZHONGLIANDA BIOPHYSICS

Rationally designed media for cell culture

This invention relates to methods for rationally designing cell culture media for use in cell cultures, e.g., cell cultures employed in polypeptide production; cell culture media designed with the disclosed methods; methods of producing a polypeptide of interest, e.g., an antibody, using such media; polypeptides produced using the methods and media disclosed herein; and pharmaceuticals compositions containing such polypeptides. The rationally designed media contain a concentration of an amino acid that is calculated for use in cell mass, a concentration of the amino acid that is calculated for use in cell maintenance, and a concentration of the amino acid that is calculated for incorporation into the polypeptide of interest. The rationally designed media may contain a baseline-adjusted concentration, A, of at least one amino acid that is calculated according to the formula A=[(M*X)+(N*P)+(Y*M*X)]*F, wherein X is the concentration of the amino acid that is used per unit of cell mass, P is the concentration of the amino acid that is used for incorporation into the polypeptide of interest per unit of polypeptide titer, M is the multiplier for the desired peak cell density of the cell culture, N is the multiplier for the desired concentration of the polypeptide of interest, Y is the cell maintenance factor; and F is the baseline factor. This rationally designed media may also be used to produce a starting cell culture medium comprising a concentration, B, of at least one amino acid according to the formula B=[A−(Z*V)]/(1−V), wherein Z is a concentration of the amino acid in the feeding cell culture medium, and V is a volume of the feeding culture medium as a proportion of the desired cell culture medium volume.
Owner:WYETH LLC

Animal rabies virus and vaccine and production method thereof

The invention discloses a method for producing animal rabies viruses and animal rabies vaccines on a large scale. By using a bioreactor, the animal rabies viruses and the animal rabies vaccines are produced by a cell micro-carrier suspension culturing system on a large scale by the following steps of: inoculating cells for preparing the vaccine into a carrier tank containing culture solution and micro-carriers to attach the cells to the micro-carriers; growing the cells on the micro-carriers until the concentration of culture solution is 5 to 20 times the inoculation concentration under a proper culturing environment; preparing virus suspension from the rabies virus to adsorb the virus suspension to the cells; replacing cell maintenance culture solution to culture the virus under the proper culturing environment; continuously culturing for 3 to 5 days and harvesting the virus solution for the first time, wherein the solution replacement ratio is 50 percent by using a semi-continuous process; continuously culturing for 9 to 11 days and harvesting and replacing the solution once every 24 hours; and mixing the harvested virus solution and the virus solution of the bioreactor, repeatedly performing freeze-thawing twice at the constant temperature of -20 DEG C, inactivating, purifying and adding adjuvants to prepare the rabies vaccine. The method has the advantages of large production scale, high single-batch yield and relatively low production cost.
Owner:PULIKE BIOLOGICAL ENG INC

Method for producing porcine reproductive and respiratory syndrome viruses (PRRSV) in large scale

The invention discloses a method for producing porcine reproductive and respiratory syndrome viruses (PRRSV) in a large scale. The method comprises the following steps of: producing PRRSV by using a cell microcarrier suspension culture system by utilizing a bioreactor; inoculating host cells used for preparing the viruses to a carrier pot containing a culture solution and microcarriers and uniformly mixing the cells and the microcarriers to ensure that the cells are attached on the microcarriers; under a proper culture environment, providing sufficient nutrients and an appropriate gas environment for the cells to ensure that the cells grow on the microcarriers to reach a concentration 5-40 times of an inoculating concentration; changing to use a cell maintenance culture solution and preparing the PRRSV into a viral suspension to ensure that the viral suspension is adsorbed on the cells; culturing the viruses under an appropriate environment; obtaining a viral solution after continuously culturing for 2-3 days; after the viral solution is inspected to be qualified, freeze thawing the obtained viral solution twice at -20 DEG C; and inactivating and purifying to obtain a PRRSV solution. The method has large production scale, high single-batch output and relatively lower production cost.
Owner:PU LIKE BIO ENG

Virus and vaccine of porcine reproductive and respiratory syndrome and preparation method of same

The invention discloses a method for preparing virus of porcine reproductive and respiratory syndrome on a large scale. In the method, the virus of the porcine reproductive and respiratory syndrome is prepared in a cell microcarrier suspension culture system by a bioreactor. The method comprises the following steps of: inoculating host cells for preparing the virus to a carrier tank containing culture solution and a microcarrier, and mixing the cells and the microcarrier uniformly to ensure that the cells are attached to the microcarrier; providing sufficient nutrients and appropriate gas environment for the cells under the appropriate culture environment to ensure that the cells are grown until the cells are in an amount which are 10 to 20 times of the inoculation concentration on the microcarrier; preparing virus suspension from the virus of the porcine reproductive and respiratory syndrome by using cell maintenance culture solution to ensure that the suspension is adsorbed to the cells; culturing the virus under the appropriate culture environment; culturing continuously for 2 to 3 days to obtain virus solution; and after the virus solution passes inspection, performing freeze thawing on the virus solution twice at the temperature of -20 DEG C, and inactivating and purifying to prepare an inactivated vaccine of the porcine reproductive and respiratory syndrome or adding a freeze-drying protective agent for freeze drying to prepare a live vaccine of the porcine reproductive and respiratory syndrome. The method has large production scale, high yield of single batch and low production cost.
Owner:PU LIKE BIO ENG

Production method of pseudorabies virus vaccine

The invention discloses a production method of a pseudorabies virus vaccine. The production method comprises the following steps: (1) adopting a cell of a continuous cell line, subjecting the cell to digestive passage, and continuously culturing the cell in a cell culture flask by using a cell growth medium; (2) diluting a virus seed into 10 times by using a cell maintenance medium, and inoculating a cell monolayer to obtain an infected cell virus liquid, that is, a virus seed for production; (3) preparing the cell monolayer formed in the step (1) into a cell suspension through digestion, inoculating the cell suspension into a bioreactor, and adding a microcarrier into the bioreactor; (4) performing virus-introduction operation on the cell, wherein the introduced virus seed is the virus seed produced in the step (2); and (5) harvesting liquid in the reactor together with the microcarrier until all of the cells on the microcarrier fall off and a dissolved oxygen value increases obviously, placing the liquid and the microcarrier under conditions of minus 20 DEG C, repeatedly freezing and thawing the liquid and the microcarrier twice, and removing the microcarrier and cell fragments to prepare the pseudorabies virus vaccine. The production method of the pseudorabies virus vaccine is short in production cycle and large in yield.
Owner:成都史纪生物制药有限公司

Method for separating and extracting stem cells from placenta, umbilical cord or adipose tissue

The invention relates to a method for separating and extracting stem cells from a placenta, umbilical cord or adipose tissue, which comprises the following process steps: firstly mixing the placenta, umbilical cord or adipose tissue and a cell maintenance fluid according to the proportion by weight of 2.5-4:1, putting the mixture into a tissue crushing barrel, adding collagenase after crushing, uniformly mixing, hatching at the temperature of 37 DEG C, filtering, adding a precipitator, sucking a supernatant fluid after settling, centrifuging, removing the supernatant fluid, adding concentrated cells into a liquid of diatrizoate sodium-ficoll 400#, then centrifuging, collecting 10-15 ml of intermediate cell layer, washing with the cell maintenance fluid, counting the collected cells, and providing the cells for clinical use when the cell survival ratio is more than or equal to 95 percent. The invention not only realizes the separation and the extraction of all stem cells from the placenta, umbilical cord or adipose tissue, but also realizes industrialized production, and enables doctors to conveniently, safely and canonically obtain the adult stem cells in clinic and use the adult stem cells for treating the diseases of patients as using medicaments, thereby solving the bottleneck problem of difficult obtainment of adult stem cells in clinic and popularizing a cell treatment technology.
Owner:NINGXIA ZHONGLIANDA BIOPHYSICS

Method for producing rabies virus antigens for animals at a large scale

The invention discloses a method for producing rabies virus antigens for animals at a large scale, which produces the rabies virus antigens at a large scale by utilizing a bioreactor by a cell microcarrier suspension culture system. The method comprises the following steps: inoculating cells for preparing the antigens into a carrier tank containing a culture solution and microcarriers to enable the cells to be attached to the microcarriers; in a proper culture environment, enabling the cells to grow on the microcarriers until the quantity of the cells is 5-40 times more than inoculum density; making rabies viruses into a virus suspension, and enabling the virus suspension to be adsorbed on the cells; culturing the viruses in the proper culture environment by using a cell maintenance culture solution; continuously culturing for 3-5 days and then harvesting a virus solution for the first time, wherein a semicontinuous process is adopted and the ratio of a changed solution is 50 percent; continue culturing for 9-11 days, and harvesting the changed solution once every 24 hours; mixing the harvested virus solution with the virus solution of the bioreactor; and carrying out freeze thawing at the temperature of -20 DEG C and inactivation purification to obtain the rabies virus antigens. The method has large production scale, high single-scale yield and relatively low production cost.
Owner:PU LIKE BIO ENG

Method for large scale cultivation in bioreactor

The invention discloses a method for large scale cultivation in a bioreactor, comprising the following steps: (1) making a recombinant H5N1 avian influenza virus adapt to MDCK cells; (2) preparing a seed lot by using the adapted virus strain as a reactor large-scale culture virus, wherein the seed lot virus generation is no less than F6, the hemagglutination titer is no less than 28, and EID50 is no less than 107.5; (3) culturing the MDCK cells by using polyester sheets as a carrier and the reactor as a basket fixed bed, and pouring the cells on every gram of the carrier, wherein the culture medium is newborn calf serum DMEM; (4) inoculating the virus on the cultured MDCK cells, washing the cells by using a virus diluent, conducting virus inoculation, after absorption, and replacing a cell maintenance media; (5) maintaining the MDCK cells and conducting virus propagation, stirring, and replacing the cell maintenance media; and (6) monitoring the hemagglutination titer and harvesting the virus, and harvesting the reactor to culture a volume virus liquid. According to the invention, the method is easy to operate and the operation is simple, the HA titer of the virus after adaption is higher than 2<7>, and the problem that the culture virus titer cannot satisfy the demand of produce vaccine in the existing culture of the recombinant H5 subtype avian influenza virus on the MDCK cells is solved.
Owner:WUHAN KEQIAN BIOLOGY CO LTD

Inactivation inspection method suitable for BVDV, IBRV, PIV3 and BRSV inactivated vaccines

InactiveCN108152498AGuaranteed infection timeImprove the detection rateMaterial analysisFreeze thawingAntigen
The invention discloses an inactivation inspection method suitable for BVDV, IBRV, PIV3 and BRSV inactivated vaccines, and relates to the field of biological medicines, in particular to an inactivation inspection method for BVDV, IBRV, PIV3 and BRSV inactivated vaccines. The inactivation inspection method comprises the following steps: (1) adsorbing an inactivated virus liquor on an appropriate cell for 40-60 min, and shaking for 2-3 times in the period; (2) after adsorption, adding a certain volume of a cell maintenance fluid in a cell spinner bottle, continuously performing culture, observing cytopathy, as for the virus fluid free from cytopathy, harvesting a cell culture fluid, and performing freeze thawing for 2-3 times; and (3) taking 10-15 ml of the frozen and thawed cell culture fluid, inoculating the taken cell culture fluid on the appropriate cell for adsorption, and continuously performing culture. Observation and fluorescence detection are performed, and if no specific fluorescence appears, complete inactivation is manifested. According to the technical scheme disclosed by the prevention, the anti-gen concentration and the virus infection time are guaranteed. The virus detection rate of the inactivation inspection method is high, and the bio-safety of the inactivated vaccines can be effectively ensured.
Owner:华威特(江苏)生物制药有限公司

Collection and enrichment system for microbial aerosol

The invention provides a microorganism aerosol collecting and enriching system, which comprises an aerosol collecting assembly, an aerosol enriching assembly and a sample collecting assembly, wherein the aerosol collecting assembly is connected to the input end of the aerosol enriching assembly, the sample collecting assembly is arranged on the outer side of the aerosol enriching assembly in a sleeving mode, the aerosol enriching assembly comprises an aerosol enriching device and a collecting tube cover, the aerosol enriching device is filled with a cell maintenance solution, the bottom end of the aerosol enriching device is open, a filter plate is packaged on the outer side of the opening, an air inlet pipe and an air outlet pipe are arranged on the collecting pipe cover in a penetrating manner, the air inlet pipe comprises a first end and a second end, the air outlet pipe comprises a third end and a fourth end, the first end and the third end are located above the collecting pipe cover, the second end and the fourth end are both located in the aerosol enriching device, the second end is fixedly connected with a porous spheroid located in the cell maintenance solution, the first end is connected with a first sucking pump, the fourth end is located above the liquid level of the cell maintenance solution, and a plug is detachably inserted into the third end.
Owner:JIANGSU PROVINCE HOSPITAL THE FIRST AFFILIATED HOSPITAL WITH NANJING MEDICAL UNIV
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