25 results about "Cellular viability" patented technology

Provided are methods for the creation of ultra-high density cryopreserved cell banks. In certain embodiments, these methods employ altered perfusion culture techniques that allow for production of ultra-high density cell cultures that can be cryopreserved at unexpectedly high cell densities without the need for any cell concentration steps, while retaining excellent cell viability and quality.

A method of inhibiting a cancer cell growth includes contacting the cancer cell with a Cu2(OH)3NO3 / CaSiO3@g-C3N4 nanocomposite material containing graphitic carbon nitride (g-C3N4), copper hydroxide nitrate (Cu2(OH)3NO3) and calcium silicate (CaSiO3), achieving an inhibition efficiency on human breast carcinoma (MCF-7) and human hepatocellular carcinoma (HepG-2) cell growth of greater than 95% in an in-vitro cellular viability assay.

The process provided in this invention transfects primary cells with gene editing reagents using a high-volume gas-permeable cell culture device and a flow-through electroporation device under conditions that improve gene editing performance, cell yield, and drug (DP) quality characteristics for cell therapy applications. As demonstrated in the examples, primary cells edited according to the process provided herein achieved improved double-strand break (DSB) formation rates, increased frequency of homology-directed repair (HR) and non-homologous end joining (NHEJ) combinations, increased frequency of bi-allelic and mono-allelic HR events, improved cell viability, proliferative capacity, and cell fitness after gene editing, and reduced manufacturing time.

The invention relates to the technical field of biomedical processing, and particularly provides a cryopreservation method of cord blood mononuclear cells, which comprises six steps of whole blood separation, plasma inactivation, CBMC cell separation, cryopreservation liquid preparation, cell cryopreservation and detection after cryopreservation recovery. The CBMC cell separation comprises three sub-steps; performing cell cryopreservation, namely averagely dividing the separated CBMC into a plurality of parts, filling the CBMC into a cryopreservation tube according to a certain density, putting the cryopreservation tube into a programmed cooling instrument, and performing cryopreservation according to a plurality of programs; and detecting after cryopreservation and resuscitation, wherein the detection comprises three sub-steps of cell viability detection, cell recovery rate detection and cell function detection. The problems that in the cryopreservation process of a traditional method, cell viability is reduced, the number of cells is greatly reduced, cell adhesion and caking are caused, cell functions are abnormal, cell phenotypes are changed after recovery, functional activity is damaged and the like are solved, the utilization rate of the CBMC is increased, and the technical scheme is obviously superior to the traditional cell cryopreservation method; the viability, the recovery rate and the function of the recovered cells are obviously improved.

The invention provides a fluorescent probe as well as a preparation method and application thereof, and belongs to the technical field of biological analysis and detection. The fluorescent probe disclosed by the invention has a structure as shown in a formula I; the invention also provides a preparation method of the fluorescent probe and application of the fluorescent probe in cell activity detection or preparation of a cell activity detection kit. The fluorescent probe provided by the invention can reflect the cell state by migrating organelles, realizes dual-channel imaging, and can meet the actual requirements of cell activity detection.

1. The name of the design product: multifunctional enzyme label instrument. 2. The use of the design product: for nucleic acid, protein concentration, enzyme activity analysis and other conventional molecular detection; signal transduction research, cell signal event active oxygen, modification detection; cell viability, apoptosis, killing and other overall level analysis. 3. The design points of the design product: in shape. 4. The picture or photo that best indicates the design points: perspective drawing.

Method for determining cell viability in a cell culture of biological cells for the biotechnological production of proteins, in particular antibodies, characterized in that a fluorescent dye selected from 4,4'-dianilino-1,1'-binaphthyl-5,5'-disulfonate (Bis-ANS) or 8-anilinonaphthalene-1-sulfonate (ANS) is used as a detection agent, and wherein the fluorescent dye is present in the cell culture at a concentration of 0.1 µM to 10 µM, the fluorescent dye selectively penetrates cells with damaged cell membranes and binds to intracellular proteins, the labeled cells are detected by excitation in the UV range and detection of the fluorescence in the green wavelength range, and the determination of cell viability is carried out using an automated imaging fluorescence microscope.

A method of treating cancer cells includes contacting the cancer cells with a CoO / CaSiO3@g-C3N4 nanocomposite material. The CoO / CaSiO3@g-C3N4 nanocomposite material includes hexagonal metal oxide nanoparticles comprising a CoO phase and a CaSiO3 phase dispersed on a matrix of g-C3N4 nanosheets. The hexagonal metal oxide nanoparticles have an average particle diameter in a range from 340 to 440 nm. The CoO / CaSiO3@g-C3N4 nanocomposite material has a percent inhibition for Human Breast Carcinoma (MCF-7) cells at least 55% inhibition in an in-vitro cellular viability assay.

The present disclosure provides, among other things, a method of cryopreserving and thawing cells that results in the thawed cells having high cellular viability and functionality post-thawing. In some embodiments, a large-scale method of cryopreserving cells is provided, the method comprising: (a) contacting the cells with a cryopreservation medium; (b) cooling the cells to −80° C. at a controlled rate to minimize latent heat of fusion; and (c) storing the cells in liquid nitrogen vapor phase, thereby cryopreserving the immune cells.

The invention relates to the field of application of radix aconiti carmichaeli polysaccharide, and provides novel application of radix aconiti carmichaeli polysaccharide, namely application of radix aconiti carmichaeli polysaccharide in preparation of medicine for treating rheumatoid arthritis. The method comprises the following steps: firstly, culturing MH7A cells in vitro, inducing the MH7A cells through monkshood polysaccharides with different concentrations, and systematically evaluating the regulation effect of the monkshood polysaccharides on the proliferation kinetics and apoptosis rate of the MH7A cells by adopting a cell activity detection kit and combining a flow cytometry analysis technology; it is proved that monkshood polysaccharides with different concentrations can inhibit MH7A cell proliferation and trigger a programmed cell death mechanism, and the monkshood polysaccharides are concentration-dependent. By dynamically monitoring the content of ROS in cells, the monkshood polysaccharide is found to be capable of reducing accumulation of ROS in MH7A cells. In addition, the phosphorylation modification level of NF-kB / P65 protein in MH7A cells and the expression level of TNF-alpha are measured through methods such as a Western Blot method and an ELISA method, and it is also proved that the monkshood polysaccharide can down-regulate the expression of the NF-kB / P65 protein and the TNF-alpha in the MH7A cells. In general, experimental results show that the monkshood polysaccharide has a strong anti-inflammatory effect on MH7A cells.

The present invention relates to a frozen cell formulation that can significantly improve the cell viability of cryopreserved cells, preferably stem cells. According to the present invention, the cell viability can be improved when thawing cells after freezing, thereby preventing a decrease in therapeutic efficiency that may occur during the storage and transportation of cell therapy agents.

A system and method for analyzing a sputum sample from a subject suspected of having lung cancer comprising obtaining a plurality of cells from the sputum sample from the subject, marking the plurality of cells with i) a plurality of cell lineage specific marker compositions, ii) a cell viability composition and iii) a tetra (4-carboxyphenyl) porphyrin (TCPP) composition; analyzing with the flow cytometer the plurality of cells marked with i-iii to obtain a subpopulation selected for cell size from the plurality of cells based upon an automatically selected bead size exclusion gate; from the cell size selected subpopulation, selecting a viable singlet population of cells using an automated non-debris gate and an automated singlets gate; from the viable singlet population of cells, obtaining flow cytometer values based upon the plurality of cell lineage specific marker compositions, the viability marker and the TCPP marker; applying a trained classifier to meta data from the subject and the flow cytometric values obtained; and generating, based upon the application of the trained classifier, a classification for the sputum sample wherein the classification is selected from a plurality of classification options comprising cancer and non-cancer.

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.

The invention discloses a cell tracking probe as well as a preparation method and application thereof. The retention time of the existing dye is relatively short (generally only several hours); the dye is not tolerant to fixation treatment, so that the dye leaks from cells, and the problem of efflux also exists; more seriously, the novel cell tracking and cell viability fluorescent dye has fluorescence, and aims to provide the novel cell tracking and cell viability fluorescent dye which is longer in retention time, does not need an additional cleaning step and is smaller in toxicity aiming at the technical problems existing in the existing main calcein orange red dye.

This invention relates to the field of human cell data analysis technology, and discloses a human cell viability data analysis system based on big data, comprising: a data acquisition module for acquiring raw experimental data and transcriptome data of human cell samples to be evaluated; a preprocessing and standardization module for preprocessing the raw experimental data and outputting a three-dimensional viability tensor; a multimodal fusion module for coupling and decomposing the three-dimensional viability tensor and transcriptome data to extract a shared sample coefficient matrix; an influence feature generation module for calculating dose-influence features, time-influence features, and viability-influence features based on the shared sample coefficient matrix and the three-dimensional viability tensor; and a prediction and interpretation module for establishing a linear regression model based on the viability-influence features and preset efficacy labels, and outputting the corresponding efficacy score for the sample to be evaluated. This invention achieves standardized integration and efficient analysis of multi-source heterogeneous human cell viability data.

This invention belongs to the field of biomedical technology, specifically relating to a method for preparing a renal-clearable optical diagnostic probe-cell conjugate and its application. The invention involves fusing a renal-clearable optical diagnostic probe with a maleimide-modified polyethylene glycol chain, followed by incubation with Traut's-pretreated cells to obtain the renal-clearable optical diagnostic probe-cell conjugate. This method is simple, time-saving, and efficient, applicable to various renal-clearable optical diagnostic probes and cell types. Furthermore, this method has minimal impact on cell viability, disease targeting ability, and ability to cross physiological barriers. In addition, the renal-clearable optical diagnostic probe-cell conjugate constructed using this method can release renal-clearable fragments in the disease microenvironment, thereby enabling early diagnosis of various diseases through urine analysis, exhibiting high sensitivity and specificity.

The invention provides a method for detecting the biological activity of GDF-11, which comprises the following steps: detecting the number of MPC-11 cells through a cell activity kit, analyzing the relationship between the dosage concentration of GDF-11 and the number of the MPC-11 cells, and calculating the half inhibitory concentration IC50, thereby evaluating the biological activity of GDF-11. The method has the advantages of large detection window, simple detection method, easily available raw materials, low cost and good result stability.

The present disclosure provides, among other things, a method of cryopreserving and thawing cells that results in the thawed cells having high cellular viability and functionality post-thawing. In some embodiments, a large-scale method of cryopreserving cells is provided, the method comprising: (a) contacting the cells with a cryopreservation medium; (b) cooling the cells to −80° C. at a controlled rate to minimize latent heat of fusion; and (c) storing the cells in liquid nitrogen vapor phase, thereby cryopreserving the immune cells.

There are provided a cell culture substrate that enables efficient spheroid formation for cells and can form spheroids having a uniform size and an arbitrary shape at a high cell viability, a method for producing the cell culture substrate, and a method of producing spheroids in which the cell culture substrate is used and a cell viability inside of the spheroids is excellent. A cell culture substrate includes a substrate and a stimulus-responsive polymer coated on the substrate, wherein the stimulus-responsive polymer is a block copolymer having a water-insoluble block segment and a stimulus-responsive block segment, and the cell culture substrate includes the following two regions (A) and (B): (A) an island-shaped region having cell proliferation properties and stimulus responsiveness and having an area of 0.001 to 5 mm2; and (B) a region adjacent to the region (A) and having no cell proliferation properties.

The invention discloses a steady-state long-term culture method of adipose-derived stem cells. The steady-state long-term culture method comprises the following steps: step 1, obtaining and pretreating adipose tissues; 2, constructing a basic culture system; step 3, regulating and controlling culture in stages; step 4, multi-dimensional steady state monitoring is carried out; 5, precise passage optimization is carried out; 6, whole-process pollution prevention and control; 7, cell quality identification; step 8, low-temperature preservation and recovery; according to the method, EGF, bFGF, TGF-beta1 and a Wnt inhibitor IWR-1 are added in stages, and an antioxidant is combined, so that long-term culture of more than 15 generations of AD-MSC is realized, the positive rate of a dry marker is always maintained to be greater than or equal to 95%, and the differentiation potential is not obviously reduced; a multi-dimensional steady-state monitoring system is established, the components of the culture medium and culture conditions are adjusted in real time, and the problems that the cell quality is not uniform and the inter-batch difference of the cell activity and the proliferation rate is smaller than or equal to 5% due to nutrient fluctuation and metabolic waste accumulation are avoided.

The invention relates to the technical field of gene therapy, in particular to a method for knocking out a Bax gene on the basis of an HEK293 cell line by utilizing a CRISPR / Cas9 technology, constructing a stable cell line expressed by foreign protein and improving the AAV packaging efficiency. The invention discloses a Bax2.6 cell strain. A pro-apoptosis factor Bax gene is knocked out from an HEK293T cell. Compared with a wild type HEK293T cell, the Bax2.6 cell strain provided by the invention shows higher cell activity under various stress conditions, the cell survival rate of the Bax2.6 cell strain is improved by at least 20% under the treatment conditions of small molecule inhibitors ZV-FMK, Nec-1, puromycin (Puro), cis-platinum (CDDP) and rotenone, and the Bax2.6 cell strain shows excellent environmental tolerance and production robustness. When the Bax2.6 cell strain provided by the invention is used for transient transfection and packaging of an adeno-associated virus (AAV) vector, the copy number of the obtained AAV virus can be up to 2 * 10 copies / mL, and the AAV packaging efficiency is improved by more than 10 times compared with that of a conventional HEK293T cell line.

The utility model discloses a kind of multi-cabin cell viability detection kits with spacing structure, it is related to kit technical field, including box and hatch, the hatch is hinged on the top of box, power switch is equipped at one side bottom end, multiple culture cabins are equipped in the box, each culture cabin is connected water bath cabin, the culture cabin bottom end has press rebound mechanism, the mechanism includes mechanism shell, spring holder, ratchet, top rod and helical groove sleeve.The utility model is by being put into the inside of culture cabin to water bath cabin, when water bath cabin is needed to be taken out, water bath cabin is pressed downward, top rod is moved downward, drives helical groove sleeve rotation, and then drive ratchet one-way rotation, spring in spring holder is compressed at this time, after button is released, spring holder releases elastic potential energy, pushes ratchet reverse rotation, top rod is moved upward by helical groove sleeve, makes water bath cabin rebound, simultaneously, mechanism shell provides support and protection for internal parts.

The invention belongs to the technical field of biological equipment, and particularly relates to a biological 3D printing visual craniocerebral tumor drug in-vitro screening model device and application thereof. The visual craniocerebral tumor drug in-vitro screening model device is mainly composed of a liquid inlet, a three-way valve, a bionic blood brain barrier model, a tumor model culture area, a culture container, a container cover, a cell culture bag, a camera and a cell activity detection electrode. The bionic blood brain barrier model and the tumor model can be realized by adopting a biological 3D printing mode; the construction of the in-vitro bionic blood-brain barrier model and the construction of the tumor model are integrated, and the method can be used for assisting in evaluating the type of the craniocerebral tumor treatment medicine and screening and evaluating the dosage of the medicine. Visualization of dynamic progress and drug screening in the rapid growth process of tumor cells is achieved, a stable and high-throughput drug screening chip is preliminarily formed through real-time monitoring of the cell activity detection electrode, and an auxiliary basis is provided for clinical treatment research of craniocerebral tumors.

The invention provides RNA (Ribonucleic Acid), shRNA (Short Hairpin Ribonucleic Acid) and a DNA (Deoxyribose Nucleic Acid) coding sequence of the shRNA for reducing the expression of a motor neuron survival gene SMN, and a delivery vector or a cell containing the RNA, the shRNA and the DNA coding sequence of the shRNA. Furthermore, the invention successfully constructs a stable transfection cell line with SMN-targeted shRNA transcriptional expression regulated by a tet-on regulation system, expression of the target gene SMN is successfully knocked down through DOX induction, a cell viability decline phenotype appears, an in-vitro cell model of the SMA disease is successfully constructed, and a better platform is provided for high-throughput screening of SMA treatment drugs. Furthermore, the invention provides a method for detecting the biological activity of the SMN protein, the method is short in period, low in cost and simple to operate, and an important means for rapidly evaluating the biological activity is provided for drugs for expressing or promoting expression of the SMN protein, including gene therapy drugs.

The present invention relates to a distributed three-dimensional cell production platform for personalized cell synthesis and treatment, in which, in cell culture and culture monitoring processes, cell culture conditions can be optimized through ultrasonic multimodal imaging and artificial intelligence analysis, and cells can be automatically cultured. Therefore, the platform according to the present invention enables a reduction in the cost required for culture by increasing the survival rate of cells during cell culture, and can improve the efficiency of cell culture.