49 results about "In vitro proliferation" patented technology

The invention belongs to the technical field of biomedicine, and relates to a human bladder cancer cell line ZSZB1 with primary cis-platinum drug resistance and application thereof. The cell line is preserved in the China Center for Type Culture Collection (CCTCC), the preservation number is CCTCC No: C2024120, and the cell line is derived from a radical operation specimen of primary chemotherapy drug-resistant bladder cancer patients of Chinese population, and has stable in-vitro multiplication capacity and primary cis-platinum drug-resistant characteristic. The ZSZB1 provides an ideal experimental model for bladder cancer pathogenesis research, cis-platinum drug resistance mechanism exploration and novel antitumor drug screening. Establishment of the cell line can effectively promote transformation from basic research of bladder cancer to clinical application, and the cell line is of great significance to promotion of precise medical development.

The invention discloses a pharmaceutical composition for inducing leukemia cell copper death and application thereof, and relates to the technical field of tumor cell apoptosis. The active ingredients of the pharmaceutical composition are oleanolic acid and illlisemom. It is found for the first time that oleanolic acid can significantly up-regulate expression of a copper death key regulatory factor FDX1, when oleanolic acid is used in combination with a copper ion carrier Ilismor, a large synergistic effect can be generated through an FDX1 / DLAT axis, leukemia cells are significantly induced to generate copper death, and therefore in-vitro proliferation and in-vivo tumor forming ability of the leukemia cells are effectively inhibited. The composition provides a new drug development strategy with a clear mechanism for treatment of leukemia, especially drug-resistant leukemia.

The invention discloses a cell culture medium supplement as well as a preparation method and application thereof, and relates to the technical field of cell biology. The supplement comprises DL-alpha-tocopheryl acetate, linolenic acid, ethanolamine, L-carnitine hydrochloride, 3, 3 ', 5-triiodoL-sodium thyrosinate, and serum albumin. As a cell culture additive with definite chemical components and simple composition, the cell culture additive can replace a traditional culture system with unknown chemical components or too complex composition, and is used for supporting proliferation, differentiation and characterization analysis of cells. The culture medium additive is clear in chemical component, simple to prepare, stable in effect and relatively low in cost, can promote in-vitro proliferation and oxidation resistance of cells including human embryonic stem cells and mesenchymal stem cells, and has important application value in actual production.

The invention discloses an application of SB216763 in improving the in-vitro proliferation capacity of porcine spermatogonium. Therefore, the SB216763 can be used as an additive of the porcine spermatogonial cell in-vitro culture medium, and the porcine spermatogonial cell in-vitro culture medium combined with various growth factors is prepared. Researches show that clone groups of the porcine spermatogonium cultured by the culture medium additionally added with the SB216763 are increased, the number of the clone groups is remarkably increased, the expression quantity of genes related to proliferation is remarkably increased, apoptosis genes are remarkably reduced, and the expression of genes related to reproductive specificity and pluripotent maintenance is also increased, so that the in-vitro proliferation and long-term maintenance of the porcine spermatogonium are improved, and the porcine spermatogonium can be used for preparing a novel culture medium for the porcine spermatogonium. Therefore, the in-vitro culture level of the PSCs is systematically improved. Therefore, the invention provides a reliable basis for establishing a long-term, stable and efficient in-vitro culture system of the porcine spermatogonium, and provides a stable and efficient cell model for gene editing of the porcine spermatogonium, preparation of transgenic animals and research on reproductive diseases; the method also has potential application value in the aspects of genetic improvement of pig breeds or preservation of male germplasm resources.

This invention discloses a jugnitin compound and its applications. The invention provides a jugnitin compound, which is a compound of formula (I), or a pharmaceutically acceptable salt, stereoisomer, or tautomer thereof; the compound provided by this invention has one or more effects selected from the group consisting of: (1) inhibitory activity against the in vitro proliferation of tumor cells; (2) in vivo tumor-suppressive effect; (3) in vivo tumor-targeting ability; and (4) good in vivo safety.

The present application relates to bovine tuberous skin disease virus, in particular, to bovine tuberous skin disease virus ORF145 gene deletion strain and its application. The gene deletion strain is characterized in that it is constructed by deleting ORF145 gene from bovine tuberous skin disease virus virulent strain LSDV XJ201901. Compared with the parent strain, the replication level of the constructed deletion strain rLSDVΔ145 in in vitro cells is equivalent to that of the parent strain, indicating that the gene deletion does not affect the in vitro proliferation ability of the virus. In addition, after rLSDVΔ145 infects MDBK cells, it can significantly reduce the mRNA expression level of pro-inflammatory factors IL-1beta and IL-6, while not interfering with the expression of IFNbeta mRNA. Further transcriptomic analysis shows that the deletion strain significantly reduces virulence and pathogenicity on the basis of maintaining good immunogenicity; it not only loses the inhibition ability to the innate immune pathway, but also can effectively activate the adaptive immune response, and significantly reduce the risk of inflammation and tissue damage, showing significant potential as a safe and efficient attenuated live vaccine.

The invention provides the application of the Chaetomugilin Q in preparation of the medicine for treating the tumors, the research proves that the Chaetomugilin Q can regulate and control the pathway of pathways in caner and the pathway of EGFR (epidermal growth factor receptor) tyrosine kinase inhibitor caner, and particularly, the Chaetomugilin Q can remarkably inhibit in-vitro proliferation of cancer cells of breast cancer, prostatic cancer, liver cancer, melanoma or leukemia, so that the Chaetomugilin Q has the application prospect in tumor treatment.

Compositions and methods are provided for modifying eukaryotic cells, e.g., to express a transgene of interest and / or to generate a population of expanded cells ex vivo.SOLUTION: A method of transducing a population of eukaryotic cells, such as a population of pluripotent cells, to express a gene of interest by contacting the cells with a viral vector, e.g., a lentiviral vector, and a diblock copolymer, e.g., a diblock copolymer comprised of a hydrophilic region and a hydrophobic region. For example, the diblock copolymer can be composed of polyoxyethylene (PEO) subunits and polyoxypropylene (PRO) subunits. In addition, the compositions and methods of the invention can be used to promote the ex vivo expansion or survival of a population of pluripotent cells (e.g., CD34 + hematopoietic stem or progenitor cells), for example, by contacting the cells with a diblock copolymer.SELECTED DRAWING: None

A hematopoietic stem cell (HSC) proliferation medium includes a basal medium, 0.1 ng / mL to 200.0 ng / mL of thrombopoietin (TPO), 0.1 ng / mL to 200.0 ng / mL of stem cell factor (SCF), 0.1 ng / mL to 20.0 ng / mL of interleukin-3 (IL-3), 0.1 ng / mL to 100.0 ng / mL of interleukin-6 (IL-6), 0.1 ng / mL to 200.0 ng / mL of Flt-3 ligand (FL), 0.1 μg / mL to 20.0 μg / mL of insulin, 0.1 μg / mL to 120.0 μg / mL of transferrin, 0.1 g / L to 3.0 g / L of human serum albumin (HSA), 0.1 μM to 4.0 μM of pyrimido-indole derivative UM729, and 0.1 μM to 2.0 μM of aryl hydrocarbon receptor (AhR) antagonist StemRegenin 1 (SR1). Methods for in vitro proliferation of hematopoietic stem cells (HSCs) and in vitro production of hematopoietic progenitor cells (HPCs) using the HSC proliferation medium are also provided.

The invention belongs to the field of immune drug development. The invention particularly relates to a T cell in-vitro proliferation culture method and application thereof. By changing the coating amount of the anti-CD3e antibody and the concentration combination of beta-mercaptoethanol with different concentrations in the T cell culture medium provided by the invention, the effect of accurately controlling the background proliferation of the T cells is achieved; the kit prepared from the related raw materials of the method is used for detecting the activity of the immune preparation.

The invention provides application of an overexpressed ifitm3 gene in promoting in-vitro steady state maintenance of oogonial stem cells of paralichthys olivaceus, and belongs to the technical field of gene engineering. The overexpression vector containing the ifitm3 gene is constructed and transfected to the oogonial stem cells of the paralichthys olivaceus, so that the in-vitro proliferation activity and migration ability of the stem cells can be remarkably enhanced, and the undifferentiated state and the stemness characteristic of the stem cells are effectively maintained. The technology provides a key means for in-vitro long-term culture, directional induced differentiation and excellent germplasm resource preservation of paralichthys olivaceus oogonial stem cells, and has important application value in fish reproductive biology research, gene editing breeding and culture industries.

The invention discloses an ecteinin compound and an application thereof. The invention provides a compound as shown in a formula (I), or pharmaceutically acceptable salt, ester, stereoisomer, tautomer, polymorphic substance, solvate, isotope label, metabolite or prodrug thereof. The compounds provided by the invention have one or more effects selected from the following groups: (1) the compounds have inhibitory activity on in-vitro proliferation of tumor cells; (2) an in-vivo tumor inhibition effect is achieved; (3) the polymer has in-vivo tumor targeting ability; and (4) the compound has good in-vivo safety.

The invention relates to the technical field of biological medicines, and particularly discloses a drug-resistant cell line MC38-IR for immunotherapy of mouse colon cancer as well as a preparation method and application of the drug-resistant cell line MC38-IR. The cell line is preserved in the China Center for Type Culture Collection, and the preservation number is CCTCC NO: C2025358. Experimental results show that the MC38-IR cell line has a remarkable PD-1 antibody drug resistance characteristic while maintaining the in-vitro proliferation and migration capabilities of parent cells. An in-vivo experiment shows that the tumor volume of a mouse inoculated with the cell line is not obviously inhibited after PD-1 antibody treatment, the lifetime has no statistical difference compared with that of a control group, and the parent MC38 cell shows obvious treatment response. The invention solves the problem of lack of colon cancer immunotherapy drug resistance models in the prior art, provides an important tool for researching drug resistance mechanisms, screening reverse drug resistance drugs and developing combined treatment strategies, and has higher scientific research and clinical application values.

The present application provides a method for in vitro proliferation of cells, comprising the following steps: adding a NKG2A conjugate to a first cell culture solution to form a second cell culture solution; and adding a cell to a cell culture container containing the second cell culture solution for a cell expansion culture to obtain an expanded cell. The method for in vitro proliferation of cells provided by the present application can efficiently obtain a large number of high-purity NK cells, and the expanded cells have a significant tumor killing ability.

The invention discloses application of TAT-MSI1 fusion protein in promotion of virus in-vitro proliferation, and belongs to the technical field of virology. The TAT and the RNA binding protein MSI1 are fused for the first time, the TAT-MSI1 fusion protein is constructed, the efficient cell penetrating power of the TAT is combined with the RNA binding protein function of the MSI1, and a brand new tool is provided for studying replication of some viruses difficult to replication; secondly, a transmembrane delivery system is prepared by using the cell-penetrating peptide TAT, so that a technical means is provided for rapidly and greatly obtaining antigens in vaccine production.

The present application relates to the technical field of cell expansion, in particular to a NK cell culture method based on multi-factor combined induction and application. The present application combines CD16, OK432, IL-2, IL-15, IL-18 and NK cell proliferation peptide and other factors to synergistically promote the proliferation of NK cells, so that the in vitro proliferation activity of NK cells is significantly improved, which is conducive to promoting the application of NK cells in clinical treatment.

The invention belongs to the technical field of medicines, and particularly relates to a pharmaceutical composition for treating pancreatic cancer and a preparation method thereof. The pharmaceutical composition comprises an annonaceous acetogenin derivative and cimicifugin in a mass ratio of 1: (0.3-0.5). The annonaceous acetogenin derivative is prepared through three-step reaction of hydrolysis, oxidative bromination and condensation, and has remarkable OGT enzyme inhibitory activity. After the annonaceous acetogenin derivative and the cimicifugin are combined according to a specific mass ratio, the in-vitro proliferation inhibition effect on pancreatic cancer cells and the tumor inhibition effect in an animal model of the annonaceous acetogenin derivative and the cimicifugin are remarkably better than those of a single drug and an original drug, and the annonaceous acetogenin derivative and the cimicifugin have a good synergistic interaction characteristic; in addition, the composition can obviously reduce the toxicity to normal cells, and provides a new drug selection and combined treatment strategy for pancreatic cancer treatment.

The invention relates to a bovine nodular dermatosis virus, in particular to a bovine nodular dermatosis virus ORF145 gene deleted strain and application of the bovine nodular dermatosis virus ORF145 gene deleted strain. The gene deletion strain is characterized in that the gene deletion strain is constructed by deleting an ORF145 gene from a bovine nodular skin disease virus virulent strain LSDV XJ201901. Compared with a parent strain, the replication level of the constructed deletion strain rLSDVdelta145 in in-vitro cells is equivalent to that of the parent strain, which indicates that gene deletion does not affect the in-vitro multiplication capacity of the virus. In addition, after MDBK cells are infected with the rLSDVdelta145, the mRNA expression level of proinflammatory factors IL-1beta and IL-6 can be remarkably reduced, and meanwhile expression of IFN beta mRNA is not interfered. Further transcriptomics analysis shows that the toxicity and pathogenicity of the deletion strain are obviously reduced on the basis of keeping good immunogenicity; the vaccine not only loses the ability to inhibit natural immune pathways, but also can effectively activate adaptive immune response and obviously reduce inflammatory response and tissue injury risks, and shows significant potential as a safe and efficient attenuated live vaccine.

The invention relates to the technical field of biological medicine, in particular to thunberg fritillary bulb polysaccharide as well as an extraction and purification method and application thereof. The extraction and purification method comprises water extraction, alcohol precipitation and two times of purification, the extraction and purification method is stable in process and good in repeatability, and a polysaccharide component with high uniformity can be efficiently obtained from the thunberg fritillary bulb. In addition, the intestinal flora structure of a human body can be remarkably regulated, specifically, the capacity of the flora for totally producing short-chain fatty acids (SCFAs, such as acetic acid, propionic acid and butyric acid) is improved, the relative abundance of beneficial bacteria (such as bifidobacteria and bacteroides) is increased, and the abundance of potential harmful bacteria is reduced; in-vitro proliferation of common probiotics (such as bifidobacterium adolescentis, bacteroides multiflora and the like) can be remarkably promoted, it is indicated that the Zhejiang fritillaria thunbergii has excellent prebiotic potential, and a new way is provided for deep development and high-value utilization of the Zhejiang fritillaria thunbergii.

The invention relates to the technical field of biological medicine, in particular to application of a COMPP-LPAR4 signal axis related to colorectal cancer. The invention finds that the expression level of LPAR4 in a colorectal cancer cell line is remarkably up-regulated, knocking-down of LPAR4 can realize in-vitro proliferation and migration of colorectal cancer cells, migration and proliferation capacity of overexpression COMP-induced colorectal cancer cells are reversed, and it is proved that LPAR4 plays a relatively important role in a COMP-regulated cancer promotion process. Targeted intervention of the signal axis can significantly weaken the migration and proliferation ability of colorectal cancer cells.

The invention discloses a synthesis method of a sulfonyl-containing indolo [1, 4] diazacycloheptanedione compound, and belongs to the technical field of organic chemistry. The method comprises the following steps: by taking N-alkyl-N-(2-(1H-indole-aryl) methacrylamide, aryl diazonium salt and a sulfur dioxide source as raw materials, carrying out heating reaction in a solvent under a catalyst-free condition, generating a sulfonyl free radical intermediate from the aryl diazonium salt and the sulfur dioxide source in situ, and carrying out free radical addition-cyclization reaction to obtain a target compound. According to the present invention, the method has characteristics of no catalyst, simple process, environmental protection, target product yield reaching up to 99%, strong substrate universality and strong functional group compatibility, and the synthesized compound has good in vitro proliferation inhibition activity on human colon cancer and breast cancer tumor cells, provides candidate molecules for anti-tumor drug research and development, and can easily achieve industrial production.

This invention belongs to the field of pluripotent stem cell culture, and relates to a culture medium additive, culture method, and culture medium for improving the proliferation rate of pluripotent stem cells. The additive is SUN11602; the additive is added to a basal culture medium with well-defined components. The technical solution of this application, while fully retaining and being compatible with existing culture medium formulations, significantly and safely improves the in vitro proliferation rate, colony formation rate, and population expansion efficiency of hPSCs by adding exogenous compounds without compromising hPSC pluripotency and genomic integrity.

The application belongs to the technical field of microorganisms, and provides GLP-1 / GIP recombinant engineering bacteria as well as a construction method and application thereof.The construction method comprises the following steps: taking a lactic acid bacteria constitutive expression plasmid as a skeleton, constructing a recombinant expression plasmid carrying a GLP-1 / GIP fusion target gene, introducing the recombinant expression plasmid into a host Lactobacillus plantarum, and obtaining the GLP-1 / GIP recombinant engineering bacteria; and the nucleotide sequence of the GLP-1 / GIP fusion target gene is shown in SEQ ID NO:1.The GLP-1 / GIP recombinant engineering bacteria not only have excellent in-vitro proliferation capacity and biological safety, but also exhibit significant effects of reducing blood sugar and weight, protecting liver and remodeling intestinal microecology in an obese mouse model.The application provides a new strategy and experimental basis for oral biological treatment of metabolic diseases, and has important theoretical value and broad clinical conversion prospects.

The invention belongs to the field of cell biology, and particularly relates to an in-vitro induction composition and an in-vitro induction culture method of cord blood-derived gamma delta T cells. The method comprises the following steps: performing induced differentiation on CD34 < + > hematopoietic stem cells to form gamma delta T cells, performing in-vitro separation and amplification on CD34 < + > to obtain a large number of hematopoietic stem cells, performing induced differentiation on the hematopoietic stem cells by using a proper induction culture medium to form gamma delta T, and performing proliferation culture through a proper in-vitro proliferation culture medium. According to the method, efficient amplification of the cord blood source gamma delta T cells can be achieved, the amplification capacity of the gamma delta T cells is remarkably improved, the problem that the amplification amount of the cord blood gamma delta T cells is insufficient is effectively solved, and the clinical use requirement can be met.

The application discloses a kind of RSP03 recombinant proteins, culture medium for prawn cell in vitro proliferation culture and its culture method, proliferation is prawn cell in vivo proliferation or prawn cell in vitro proliferation;The nucleotide corresponding to RSP03 recombinant protein is as shown in sequence SEQ ID NO.1, and the amino acid sequence of RSP03 recombinant protein is as shown in SEQ ID NO.2.The endogenous growth factor RSP03 gene of South American white prawn is cloned, and expression vector is constructed to express and purify endogenous LvRSPO3 recombinant protein, and the in-vitro culture activity of prawn cell can be extremely significantly enhanced by adding 5~30 ng / mL LvRSPO3 recombinant protein on base medium, can more effectively promote the proliferation and division of prawn cell in vitro, and can be cultured for at least 30 days, to overcome the problem that prawn cell is difficult to divide when in vitro culture, and provide a train of thought for establishing prawn continuous cell line.

The invention discloses an RSPO3 recombinant protein, a culture medium for in-vitro proliferation culture of prawn cells and a culture method of the RSPO3 recombinant protein. The proliferation is in-vivo proliferation of the prawn cells or in-vitro proliferation of the prawn cells; the nucleotide corresponding to the RSPO3 recombinant protein is as shown in a sequence SEQ ID NO.1, and the amino acid sequence of the RSPO3 recombinant protein is as shown in a sequence SEQ ID NO.2. An endogenous growth factor RSPO3 gene of penaeus vannamei boone is cloned, an expression vector is constructed to express and purify an endogenous LvRSPO3 recombinant protein, and 5-30 ng / mL of the LvRSPO3 recombinant protein is added to a basic culture medium, so that the in-vitro culture activity of penaeus vannamei boone cells can be remarkably enhanced, in-vitro proliferation and division of the penaeus vannamei boone cells can be more effectively promoted, and the survival rate of the penaeus vannamei boone cells is increased. The method can be cultured for at least 30 days for a long time, and provides a thought for overcoming the problem that prawn cells are difficult to split during in-vitro culture at present and establishing a prawn continuous cell line.

The present application belongs to the field of cell culture. The present application provides a method for inducing, culturing and amplifying human natural killer cells, comprising the following steps: extracting peripheral blood from a healthy person, and separating PBMC; coating a container with an NK activation solution; culturing with an NK cell proliferation medium, inoculating PBMC, and incubating at a constant temperature for 1-6 days; and proliferating with an NK cell amplification medium, and incubating at a constant temperature for 1-10 days. The present application effectively improves the in-vitro proliferation rate of human natural killer cells. Through flow cytometry analysis, the proportion of NK cells with phenotypes of CD3, CD16 and CD56 in the cell population is all above 80%, and the cell phenotype is CD56 + CD3 ‑ NK cell group, and the positive rate is 97.57%. The present application has a simple composition formula, ideal culture effect, and wide application prospect.

The invention discloses a mesenchymal stem cell with anti-apoptosis and in-vivo tracing functions as well as preparation and application thereof, and relates to the technical field of biology. The invention specifically discloses a mesenchymal stem cell for overexpressing anti-apoptotic protein, luciferase and fluorescent protein. The mesenchymal stem cell surface marker retains the characteristics of the mesenchymal stem cell surface marker, has multidirectional differentiation potentials of adipogenesis, osteogenesis and cartilage formation, and has no tumorigenicity in immunodeficient animal bodies; the method can track the cell distribution and survival ability in real time, has a good application prospect in the aspects of treating neurodevelopmental disorders or neurodegenerative diseases, researching a treatment mechanism and in-vivo / in-vitro proliferation, migration, colonization, differentiation or integration behaviors of the neurodevelopmental disorders or neurodegenerative diseases, provides a standardized and visual research platform for the field of stem cell research, and has a wide application prospect. The important scientific value and clinical transformation potential are realized.

The invention discloses a recombinant Marek's virus vector vaccine strain for expressing H9N2 subtype avian influenza virus HA protein and application of the recombinant Marek's virus vector vaccine strain, and belongs to the technical field of veterinary biological products. The amino acid sequence of the HA protein expressed by the recombinant Marek's virus vector vaccine strain is as shown in SEQ ID NO. 1. The amino acid sequence of the HA protein is obtained by modifying the 84th amino acid of the amino acid sequence shown as SEQ ID NO.2 into tryptophan from serine, eliminating the 82nd glycosylation site of the HA protein, replacing the signal peptide of the HA gene with the Ig lambda signal peptide of chicken, and adding GGGGS linker and one tetanus toxin T cell epitope to the carboxyl terminal of the HA protein. The vaccine strain is good in in-vitro proliferation characteristic, stable in genome heredity and capable of stably expressing HA protein, no side reaction or pathogenic effect exists after SPF chicken is immunized, 100% protection against H9N2 strain attack can be provided 21 days after immunization, and the problem that the immune protection blank period of other vaccines such as H9N2 inactivated vaccines is long is solved.

The invention discloses an application of pristimerin in preparation of a medicine for treating acute myelogenous leukemia, it is found for the first time that pristimerin acts on targets CTNNB1 and HIF1A of acute myelogenous leukemia cells and has an inhibition effect on acute myelogenous leukemia, CTNNB1 and HIF1A serve as the targets, high expression of CTNNB1 and HIF1A is regulated, and the medicine for treating acute myelogenous leukemia is obtained. The traditional Chinese medicine composition can be used for treating acute myelogenous leukemia, inhibiting abnormal proliferation of leukemia cells, promoting apoptosis of acute myelogenous leukemia cells and retarding the cell cycle of acute myelogenous leukemia at G0 / G1 phase, so that the tumor microenvironment is improved, the dominant conditions of leukemia cell survival are broken, and the purpose of treating acute myelogenous leukemia is achieved. The pristimerin can be used for accurately and effectively inhibiting in-vitro proliferation of acute myelogenous leukemia cells, a new thought and a new direction are provided for development of drugs for diagnosing and treating acute myelogenous leukemia, and the application range of pristimerin in the field of AML treatment is widened.