65 results about "Viable cell" patented technology

The present disclosure includes compounds, compositions, and methods for nucleic acid amplification reactions. In particular, the present disclosure provides compounds, compositions, and methods for viability PCR (vPCR) applications, in which the compounds that selectively bind to nucleic acids from non-viable cells.

Methods for distinguishing particles in a fluid sample are disclosed. In one embodiment, the method includes acquiring a background SIMI image and a background brightfield image of a membrane filter while the membrane filter is free of a fluid sample, introducing a fluid sample onto the membrane filter, acquiring a SIMI image and a brightfield image of filtered particles resting on the membrane filter, distinguishing between the filtered particulates and the membrane filter based on the background SIMI image, generating a particle mask based on the SIMI image, and detecting beads via the particle mask. Methods for distinguishing particulates include distinguishing between viable and non-viable cell populations, distinguishing between cellular and non-cellular particulates, distinguishing between biological and non-biological particulates, distinguishing between first and second protein types, determining stability of monoclonal antibody drugs, identifying beads in a cell therapy, and detecting bacteria.

The application discloses a high Bacillus altitudinis SW2, belongs to the field of degrading bacteria screening, and the high Bacillus altitudinis SW2 has been preserved in the China General Microbiological Culture Collection Center on May 8, 2024, has a preservation number of CGMCC No.30525, and has a preservation address of No.3, Xili, Beichen West Road, Chaoyang District, Beijing City; is classified and named as the high Bacillus altitudinis. The strain is obtained through screening, purification and identification technologies from grape branch compost, and through research on strain characteristics, fermentation conditions and cellulase activity of the strain, it is shown that the enzyme activity of the strain is 547.82 U / g under the condition of 30 DEG C and PH=7, and when 5% casein is added as a protective agent, the effective viable cell number of the high Bacillus altitudinis SW2 is the highest and can reach 1.73*10 14 cfu / g.

Materials and methods for controlling nutrient feed in cell culture processes are provided. A sample is received from a bioreactor comprising a cell culture. A viable cell density and a residual nutrient measurement are determined from the received sample. A daily nutrient feed target is calculated based on the viable cell density and the residual nutrient measurement. The nutrient is fed to the bioreactor according to the calculated daily nutrient feed target.

The application belongs to the technical field of microorganism detection, and particularly relates to a bifidobacterium viable count detection culture medium, a detection method and application. The application provides a culture medium for detecting bifidobacterium, wherein the culture medium comprises MRS culture medium and an antibiotic solution; the antibiotic solution comprises streptomycin, kanamycin and sodium propionate. The culture medium can ensure normal growth of bifidobacterium, has strong inhibition on lactobacillus, can effectively realize accurate detection of bifidobacterium, and avoids interference of lactobacillus.

The invention belongs to the technical field of microorganisms, and discloses a high-density fermentation process of vagina-sourced lactobacillus crispatus. The invention discloses a fermentation medium which comprises glucose, tryptone, beef extract, yeast extract, dipotassium phosphate, ammonium citrate, L-cysteine or salt thereof, magnesium sulfate and manganese sulfate. According to the fermentation culture medium provided by the invention, through scientific compounding of a carbon source, a nitrogen source and key factors, the final viable concentration and thallus activity of vagina-derived lactobacillus crispatus can be remarkably improved. The fermentation culture medium is adaptive to the micro-aerobic metabolism characteristics of vagina-derived lactobacillus crispatus by precisely regulating and controlling the dissolved oxygen level (initial dissolved oxygen is controlled to be 5% or below), and when the fermentation culture medium is used for culturing lactobacillus crispatus, the fermentation delay period can be remarkably shortened to 2-3 h, the final viable bacterium yield is greatly increased to 1 * 10 < 10 > CFU / mL or above, and the production cost is reduced. And an efficient and stable technical support is provided for industrial production of the probiotic preparation.

Provided are methods of selecting a transformed cell, including methods comprising delivering an inhibitor of a protein to a plurality of cells, exposing the plurality of cells to a compound, wherein the protein (if present and active) converts the compound to a toxic compound, and selecting viable cells from the plurality of cells not killed by exposure to the compound.

Described are methods and systems for enhanced dissociation of cells from a solid biological tissue sample. In some embodiments, a self-contained cartridge apparatus includes a first chamber for receiving the tissue sample, enables ultrasonic energy from a transducer assembly of a processing unit to dissociate cells from the sample in the first chamber, and collect viable cells of interest from an aqueous suspension in a second chamber fluidly connected to the first chamber via a channel. In some embodiments, to enhance dissociation of viable cells, a filter device includes a tubular body configured to be telescopically inserted into a container containing the tissue sample in an aqueous fluid. The filter device also includes a cell-filter mesh that covers a bottom opening of the tubular body and that is configured to compress the tissue sample to expel cells from the sample when the filter device is fully inserted into the container.

The invention provides a complex microbial inoculant for promoting acid sugarcane soil improvement as well as a preparation method and application thereof, and belongs to the technical field of agricultural microorganisms. The complex microbial inoculant comprises bacillus amyloliquefaciens N15300, bacillus megatherium 2020055 and bacillus licheniformis 2020049, wherein the ratio of the effective viable count of the bacillus amyloliquefaciens N15300 to the effective viable count of the bacillus megatherium 2020055 to the effective viable count of the bacillus licheniformis 2020049 is (1.8-2.2): (1.8-2.2): (0.8-1.2), and the total effective viable count is greater than or equal to 1 * 10 The complex microbial inoculant can effectively act on acidic sugarcane soil in places such as Guangxi, has multiple functions of dissolving phosphorus, dissolving potassium and secreting cellulase and protease, and can activate fixed nutrients in the soil, improve the utilization rate of a fertilizer, neutralize the acidity of the soil and improve the pH value of the soil. Pot experiments show that the bacterial agent can significantly promote sugarcane root development and plant growth, improve the fertilizer utilization rate, and provide an effective way for realizing high and stable yield of sugarcane and green improvement of acid soil.

The application discloses a safe, efficient and low-cost cleaning solution for suspended cells and application thereof, which comprises multiple kinds of glucose, amino acid supplements, vitamin supplements, anti-shear protective agents and sodium bicarbonate, is designed for the harvesting and washing process of suspended cells, can effectively remove residual culture medium, cell fragments, metabolic waste and process-related impurities while effectively maintaining cell viability, viable cell density and normal morphology, so as to realize the harvesting target of high-purity, high-activity and low-risk material of suspended cells, and is especially suitable for the production process of high-demand cell products such as cell culture meat. The cleaning solution system can be adapted to the continuous harvesting process of a large-scale bioreactor, can be integrated with automatic harvesting equipment, realizes stable, efficient and repeatable cell collection and washing operation, and meets the full-scene requirements from laboratory research and development to industrial production.

Certain embodiments of the invention are directed to evaluating and identifying cells by recording and interpreting a time-dependent signal produced by unique cell respiration and permeability attributes of isolated viable cells. Some methods comprise dividing the sample into two or more sub-samples or sample portions, mixing each sub-sample or sample portion with one or more reagents and / or one or more reactants forming distinct sub-sample or sample portion mixtures, compartmentalizing each of the sub-sample or sample portion mixtures into a plurality of small volume compartments, monitoring characteristics of the small volume compartments over time and collecting compartment data, and transmitting the collected data to at least one neural network.

The invention relates to a preservative as well as a preparation method and application thereof. The preservative comprises a bacillus velezensis microbial inoculum and an elicitor cooperating with the microbial inoculum, and the addition amount of the elicitor is 0.01-0.1% of the total mass of the preservative. The preservative further comprises at least one of a protective agent, a preservative and an antioxidant; the bacillus velezensis microbial inoculum comprises the following components in parts by mass: 20-80 parts of a bacillus velezensis microbial inoculum, 0.001-0.1 part of an elicitor, 1-25 parts of a protective agent, 0.05-1 part of a preservative and 0.05-1 part of an antioxidant. The effective viable count in the bacillus velezensis microbial inoculum is greater than or equal to 10 < 9 > CFU / mL; the bacillus velezensis microbial inoculum comprises at least one of bacillus velezensis strains LTS-AF1, LTS-AF2, LTS-AF3 and LTS-AF4, and the bacillus velezensis microbial inoculum comprises at least one of bacillus velezensis strains LTS-AF1, LTS-AF2, the strain action effect is improved, the action time is prolonged, and the strain activity is stabilized.

The invention relates to a pesticide-fertilizer granule and a preparation method thereof, the granule is composed of a compound functional flora domesticated by saline-alkali gradient stress, an organic carrier prepared by fermenting agricultural and forestry wastes and livestock and poultry manure, and a chemical synergistic component, and the effective viable count is greater than or equal to 0.5 billion CFU / g. The preparation method comprises the following steps: (1) screening salt-dissolving, growth-promoting and stress-resistant composite flora from a saline-alkali soil sample; (2) performing high-temperature aerobic fermentation on forestry and agricultural residues and livestock and poultry manure to prepare a carrier; (3) inoculating the functional flora to the carrier by adopting a segmented inoculation process; and (4) extruding and granulating at a low temperature lower than 0 DEG C. The bottleneck that a traditional bacterial fertilizer is single in function, high in carrier cost, large in granulation viable bacterium loss and the like is broken through, a chemical modifier and bacterial fertilizer synergistic application mode is innovated for severe saline-alkali soil, the soil salt content is reduced by 25%-30%, the crop yield is increased by 15%-20%, the strain survival rate is larger than or equal to 75%, and a standard and low-cost solution is provided for green improvement of the saline-alkali soil.

The invention discloses a fermentation method and application of bacillus laterosporus, and belongs to the technical field of microbial fermentation engineering and biological control. In order to solve the technical problems of unstable fermentation level, low spore formation rate and the like of bacillus laterosporus, the invention provides a high-density fermentation method of bacillus laterosporus, which comprises the following steps: adding a nutrition-signal composite inducer in stages in the middle and later periods of fermentation, and coupling pH / DO two-stage control, so that the spore formation rate is greater than or equal to 95%; besides, fermentation liquor of bacillus laterosporus and the bentonite-trehalose-sodium glutamate composite carrier are mixed and then subjected to low-temperature spray drying, the viable bacterium yield of the prepared microbial agent is larger than or equal to 95%, the viable bacterium rate is larger than or equal to 90% after the microbial agent is stored for 12 months at the temperature of 25 DEG C, and the microbial agent shows a remarkable effect in tomato root-knot nematode prevention and treatment of livestock and poultry diarrhea and has good application prospects. The compound can completely or partially replace chemical pesticides and antibiotics, and has a wide market prospect.

Provided herein are methods of culturing a mammalian cell in a liquid medium including poloxamer-188 at a concentration of 1.8 g / L or at a greater concentration than 1.8 g / L more or a liquid medium that includes a poloxamer-188 concentration that is selected based on one or more factors selected from the group of: pore size, pore type, gas flow rate, viable cell density in the medium, and markers related to cell stress.

Provided herein is a composition comprising a hydrogel covalently bound to a functional moiety, wherein the hydrogel comprises a cross linkable biopolymer and a first cell population; the cross linkable biopolymer is at least partially crosslinked by a crosslinking agent within the hydrogel; and the functional moiety has a binding affinity to a cell. Furthermore, a method of growing cells using the composition of the invention, and a method of manufacturing the composition using a kit disclosed herein are also provided.

Embodiments of the present disclosure provide compositions and methods for injectable recombinant protein-based hydrogels for therapeutic delivery. Embodiments of the composition and methods comprise a recombinant protein, a cell consortia, and a biocompatible medium. The recombinant protein comprises one or more self-association domains and a flexible linker comprising a pseudorepeat of amino acids, forming stable hydrogels in contact with the cell consortia. Such composition and methods have broad application in delivery of viable cells to a biological tissue for tissue repair and regeneration.

This provides a continuous culture process for culturing populations of shear-sensitive cells. [Solution] A continuous culture process for culturing a population of cells consisting of shear-sensitive cells, comprising controlling the gas outflow rate of a bioreactor system so that the gas outflow rate does not exceed 20 m / s, wherein the bioreactor system includes at least 25 L of culture medium, and the population of cells is 20 × 10 6 From cells / mL to 15 × 10 7 The process is characterized by achieving a steady-state viable cell concentration within the range of cells / mL, and the gas efflux rate being controlled at least until the bioreactor system reaches steady-state conditions.

In certain embodiments, this disclosure provides novel methods of increasing the viable cell density of an N−1 large-scale bioreactor cell culture, comprising culturing a host cell expressing a recombinant poly peptide of interest in a non-perfusion-based culture system, and wherein the viable cell density is increased to at least 5×106 cells / mL. In certain embodiments, the disclosure provides novel methods for large-scale production of a recombinant polypeptide of interest, comprising: (1) culturing a host cell expressing a recombinant polypeptide of interest in an N−1 stage in a non-perfusion-based culture system, wherein the viable cell density is increased to at least 5×106 cells / mL; and (2) culturing N fed-batch production cells in an enriched media with high-seed density at least 1.5×106 cells / mL, wherein the N fed-batch production cells are inoculated from the N-1 stage in a non-perfusion-based culture system.

The invention relates to the technical field of biomedical detection, in particular to a method for separating circulating tumor cells and clusters thereof. The micro-fluidic chip based on the bionic vortex turbulent flow hole array and the detachable double filter membranes and the separation method of the circulating tumor cells and the clusters thereof have the advantages that the recovery rate is high (the CTC single cell gt, 90% of the cluster gt and 85% of the cluster gt); the purity is high (the leukocyte removal rate gt is 98%); synchronously and independently separating a single CTC and a heterogeneous cluster (such as a CTC-CTC cluster and a CTC-immune cell cluster); and the cell activity is maintained (the living cell rate is greater than or equal to 85%).

The present invention provides a continuous upstream manufacturing process for the production of bispecific antibody products, which comprise at least two binding domains. The process comprises at least the steps of (i) providing in a perfusion bioreactor at least one mammalian cell culture, which is capable of expressing the bispecific antibody product, (ii) growing the mammalian cell culture at a first perfusion rate until a set point viable cell density is reached, and (iii) maintaining perfusion culture at a second perfusion rate, wherein the bispecific antibody product concentration in the bioreactor is kept below a threshold value. The bispecific antibody product is then subject to subsequent downstream processing. Moreover, the invention provides a bispecific antibody product produced by the continuous upstream manufacturing process.

Provided are a decellularized amniotic membrane matrix hydrogel and its preparation method, which includes the following steps: pre-treatment of an amniotic membrane tissue, washing, decellularization, grinding, freeze-drying, digestion, and acid-base neutralization processes. The decellularized amniotic membrane matrix hydrogel provided in this disclosure retains components such as collagen, fibronectin, and glycoproteins from the amniotic membrane while removing viable cells, thereby reducing its immunogenicity at the source. The decellularized amniotic membrane matrix hydrogel may mimic the microenvironment in which cells exist in vivo, providing the necessary support and signals for cell adhesion and growth. It can be used to construct tissue engineering scaffolds, supporting directed cell growth and tissue regeneration. Furthermore, the decellularized amniotic membrane matrix hydrogel may also serve as a drug carrier for in vivo drug release and therapy, demonstrating significant potential for application in medical research and clinical settings.

This invention discloses a method for sensitively detecting water toxicity using CHO-k1 cell fluorescence and its application, belonging to the field of water safety detection technology. The method uses CHO-k1 cells as the test carrier. After cell preparation, solid-phase extraction enrichment of water samples, and exposure to toxic substances, the method simultaneously detects cell inhibition rate and intracellular reactive oxygen species (ROS) fluorescence intensity. The total fluorescence intensity is corrected by the number of surviving cells, and the relative ROS fluorescence intensity and oxidative stress impact rate (R) of a single cell are calculated to establish a four-level toxicity grading standard, enabling quantitative evaluation of water sample toxicity. The detection sensitivity of this method is far higher than traditional cytotoxicity detection methods. It can detect sub-lethal damage from low concentrations of pollutants, and the results are accurate and reliable. The operation is standardized and applicable to rapid toxicity screening and water environment health risk assessment of various water samples, including surface water, groundwater, municipal water supply, industrial wastewater, and reclaimed water.

Methods for distinguishing particles in a fluid sample are disclosed. In one embodiment, the method includes acquiring a background SIMI image and a background brightfield image of a membrane filter while the membrane filter is free of a fluid sample, introducing a fluid sample onto the membrane filter, acquiring a SIMI image and a brightfield image of filtered particles resting on the membrane filter, distinguishing between the filtered particulates and the membrane filter based on the background SIMI image, generating a particle mask based on the SIMI image, and detecting beads via the particle mask. Methods for distinguishing particulates include distinguishing between viable and non-viable cell populations, distinguishing between cellular and non-cellular particulates, distinguishing between biological and non-biological particulates, distinguishing between first and second protein types, determining stability of monoclonal antibody drugs, identifying beads in a cell therapy, and detecting bacteria.

Provided herein are methods and materials for labeling and annotating living cells in minute organoid spheres (MOSs), as well as methods for assessing MOS containing labeled living cells.

This application provides an agent or a composition comprising viable cells or dead cells of the Lactobacillus plantarum strain (International Accession No. NITE BP-03418) as an active ingredient for preventing enteritis or necrotic enteritis, for suppressing inflammation due to enteritis or necrotic enteritis, for suppressing decreased feed conversion efficiency or body weight loss, or for suppressing infection with enteric viruses in a bird (for example, a chicken), or a method that comprises feeding a bird (for example, a chicken) with the agent or the composition.

The present disclosure provides, among other things, continuous culture methods for producing a cell product, e.g., a recombinant protein, e.g., a glycoprotein, e.g., an antibody agent or a fusion protein. In some instances, methods herein allow large-scale production of a recombinant protein using continuous culture methods. The present disclosure identifies and addresses a problem with current continuous cell culture techniques in that at large-scale culture of certain cells have insufficient viable cell concentrations and impaired cell viability. The present disclosure provides, in part, methods and systems for large-scale continuous culture of shear-sensitive cells.

Methods of improving the efficiency of production of a protein product of interest in mammalian cell culture are presented. In particular, the methods result in an increase in the quantity of a protein product produced, or decreases protein product production time in a manufacturing-scale bioreactor cell culture. The disclosed methods comprise: (a) culturing the N-1 bioreactor culture to high viable cell densities; and (b) seeding the production bioreactor culture at high viable cell seeding densities.

The present invention belongs to the field of genetic engineering technology, and in particular to a RNA polymerase β subunit mutant and its application. The RNA polymerase β subunit mutant is obtained by deleting 6 amino acids from positions 215 to 220 based on the wild-type RNA polymerase β subunit (RpoC, NCBI Reference Sequences: NP_418415.1), and has the amino acid sequence shown in SEQ ID NO.1. It is applied to Escherichia coli producing 2'-fucosyllactose and 3-fucosyllactose, which can greatly increase the number of viable cells of the strain during fermentation, improve the tolerance of the strain in the fermentation environment, and the yield of 2'-fucosyllactose or 3-fucosyllactose is also greatly improved.

Provided in the present application are a real-time quantitative detection method for next-generation probiotics on the basis of flow cytometry and the use thereof. The next-generation probiotics comprise Akkermansia muciniphila and / or Bacteroides fragilis. In the present application, flow cytometry is used in the detection of next-generation probiotics to provide quantitative data of viable and non-viable cells in real time, thereby rapidly completing the high-throughput detection of viable cells in a sample while reducing the risk of contamination during the detection process. Moreover, the detection method can also detect next-generation probiotics that are unculturable due to insufficient metabolic activity, offering rapid detection speed and high accuracy.