91 results about "Animal cell line" patented technology

The present invention relates to the use of interferon in the in vitro cultivation of animal circular ssDNA virus such as Porcine Circovirus 2 or human TT virus in an animal cell line. Increased titres of animal circular ssDNA virus are obtained by addition of interferons or agents which ensure the production of endogenous interferons by said cell line.

The present invention discloses a strain of a recombinant cell line for stably expressing classical swine fever virus E2 protein, and applications of the recombinant cell line in preparation of subunit vaccines and diagnosis reagents of classical swine fever, wherein specifically the recombinant cell line is BCSFV-E2, is preserved in the China General Microbiological Culture Collection Center, and has the preservation number of CGMCC No.7719. The classical swine fever subunit vaccine prepared by using the recombinant cell line has characteristics of high safety, good immunization effect, easy mass production, less being susceptible to exogenous virus pollution or influence of antibodies, and no influence of the maternal antibody on immunization of swine, and can induce and produce high level classical swine fever virus neutralization antibodies after the swine is immunized. In addition, the present invention further discloses a method for constructing the recombinant mammalian cell line, a method for preparing the classical swine fever subunit vaccine, and applications of the antigen expressed by the recombinant cell line in preparation of classical swine fever prevention vaccines and diagnosis reagents.

Disclosed herein is an inducible high-expression cassette comprising a dihydrofolate reductase (DHFR) promoter from which GC-rich repeat sequences are partially or entirely removed, the cassette capable of more effectively improving a gene amplification system. Also disclosed are an expression vector comprising the inducible expression cassette and optionally a gene encoding a recombinant protein of interest, an animal cell line transformed with the expression vector, and a method of mass producing and purifying a recombinant protein by culturing the transformant. The present invention enables the shortening of the time required to establish a cell line producing a recombinant protein of interest at high levels using a low concentration of a DHFR inhibitor, thereby allowing more effective production of the recombinant protein.

The invention belongs to the field of micro-organism animal cell line and relates to a human hepatoma cell line which can emit high-intensity red or green fluorescence and has high transferring ability of lung and lymph node metastasis, and a method for establishing the same. The method comprises the following steps: using the human hepatoma cell line HCCLM3 and HCCLM6 which have high transferring ability of the lung and lymph node metastasis as mother cells, performing cotransfection on plasmid DNA of 239 cells through slow virus packaging plasmids to obtain false slow virus particles by expressing red or green fluorescent protein genes through eucaryon, and infecting liver cancer cell strains of the mother cells to obtain the chromosome integrated hepatoma cell line which has high transferring ability of the lung and lymph node metastasis and can stably expressing the red or the green fluorescence. The human hepatoma cell line which has high transferring ability of the lung and lymph node metastasis in vitro can be applied to the tracer studies on tumor cells, the molecular mechanism studies on the recurrence and transferring of liver cancer, as well as the pre-clinical drug efficacy studies on new anti-tumor drugs, thus the human hepatoma cell line has wide application prospect.

The invention belongs to the field of microbial animal cell lines, and relates to a new human acute B lymphocytic leukemia cell strain CHH-1. The in vitro isolate of the mononuclear cell of the marrow of a patient with firstly-diagnosed acute B lymphocytic leukemia undergoes cell primary culture to obtain the highly aggressive human acute B lymphocytic leukemia cell strain with add(11)(q23) chromosome abnormality, the preservation number of the cell strain is CGMCC No.11797, and the highly aggressive human acute B lymphocytic leukemia cell strain with add(11)(q23) chromosome abnormality is named as human acute B lymphocytic leukemia cell strain CHH-1, has a same clone source with patient's leukemia cells, can be infinitely and stably passed in vitro, and has the characteristics of clonality add(11)(q23) genetic abnormality, high tumorigenicity and high aggressiveness. The new human acute B lymphocytic leukemia cell strain CHH-1 provides a new good cell model for researches of human acute lymphocytic leukemia with No.11 chromosome long arm structure abnormality and development of biomedicines, and has wide prospect and great practical values in revelation of the pathogenesis of the human acute lymphocytic leukemia and development of medicines.

The invention discloses a staphylococcal enterotoxin gene engineering reshaped antibody and its preparation method and use. The staphylococcal enterotoxin gene engineering reshaped antibody has an amino acid sequence shown in the formula of SEQ ID No.1 in the sequence table. Through construction of light and heavy chain eukaryotic co-expression vectors of the staphylococcal enterotoxin monoclonal antibody, a high-efficiency expression and stable-secretion mammalian cell line and the gene engineering reshaped antibody having high singularity and strong affinity are obtained. The staphylococcal enterotoxin gene engineering reshaped antibody can be used in staphylococcal enterotoxin detection, cell indirect immunofluorescence detection and flow cytometry detection.

The invention discloses a staphylococcal enterotoxin micromolecule antibody and its preparation method and use. The staphylococcal enterotoxin micromolecule antibody has an amino acid sequence shown in the formula of SEQ ID No.1 in the sequence table. The preparation method comprises the following steps of designing and constructing eukaryotic mono-promoter co-expression vectors of staphylococcal enterotoxin monoclonal antibody light chain and heavy chain variable region genes, introducing cysteine residues to C- ends of the light chain and the heavy chain to form interchain disulfide bonds by an intracellular peptide fragment self-assembling principle, and simulating antigen binding domain spatial conformation of natural antibodies to obtain high-efficiency expression stable-secretion mammal cell line and the high-singularity strong-affinity micromolecule antibody. The staphylococcal enterotoxin micromolecule antibody can be used for staphylococcal enterotoxin detection, cell indirect immunofluorescence detection and flow cytometry detection.

The invention belongs to the field of microorganism animal cell lines, and particularly relates to a pancreatic cancer cell line with lymphatic channel high migration activity. According to the pancreatic cancer cell line with the lymphatic channel high migration activity, a human pancreatic cancer BxPC-3 cell line is adopted as source cells, by means of a nude mouse palmula subcutaneous vaccinization method, lymphatic metastasis oncocyte sunlines with different migration potentialities are separated from matrilineal BxPC-3 cells, a pancreatic cancer cell line BxPC-3-LN with the lymphatic channel high migration activity is screened out through experiments inside and outside human bodies, a storage number is CGMCC NO.5421, and the storage date is October 28th, 2011. The pancreatic cancer cell line with the lymphatic channel high migration activity is the same as the matrilineal BxPC-3 cells in the genetic background, is characterized by being strong in invasiveness, high in migration efficiency and concentrated in migration, provides a suitable experimental platform for research on pancreatic cancer migration mechanism and the screening of antineoplastic drugs, and has very important significance and wide application prospects.

The present invention relates to the use of interferon in the in vitro cultivation of animal circular ssDNA virus such as Porcine Circovirus 2 or human TT virus in an animal cell line. Increased titres of animal circular ssDNA virus are obtained by one or more of the following conditions: addition of interferons or agents which ensure the production of endogenous interferons by said cell line, reduction of endosomal-lysosomal system acidification, and cholesterol depletion.

The invention belongs to the filed of microorganism animal cell lines and provides a Chinese lung adenocarcinoma cell line with high metastases potentiality of bone, lever and adrenal gland. According to the invention, hydrothorax of an adenocarcinoma first-diagnosis patient is used as primary culture of cells; the human adenocarcinoma cell has the preservation number of CGMCC No.3137 and classified name as human lung adenocarcinoma cell line CPA-Yang3, can grow adhered to the wall and has the tumorigenesis rate up to 100 percent; the human lung adenocarcinoma cell can grow quickly and metabolize vigorously; the expression levels of cancer genes EMS1, VEGF-C, IL-6, IL-8, SVIL and AR gene are higher than the expression level of SPC-A-1 cell; and the lung adenocarcinoma cancel cell has the biological characteristics of mainly transferring the bone and having high transfer potential of the lever and the adrenal gland, high growth speed of bone transfer cells and complete cell morphology. The invention can be used for providing reference data to early diagnosis for transferring the human lung adenocarcinoma bone, further establishing the relevant gene chip technology and assessing and exploring the curative effects of various medicaments and the like by comprehensively applying the gene chip, quantifying PCR (Polymerase Chain Reaction), western blot in real time and other technologies.

The invention belongs to the field of microorganism animal cell line, and in particular relates to an engineering strain containing an integron. The engineering strain is eschrichia.coli, with a preservation number of C6MCC No.2353. The strain is named DHS, and the chromosome DNA of the strain has a structure of a sequence 1. In the invention, a transposon is used as a tool to insert an integron having a known sequence into the chromosome DNA of a colon bacillus. Tests show that the strain can capture an addA2 drug-resistance gene cassette in the presence of a high-expression integrase. The establishment of the strain lays a foundation for deeply researching a mechanism for the integron to capture the drug-resistance gene cassette in strains with known genetic background, and the strain can be used as a tool strain in other researches about the integron.

The present invention relates to the use of interferon in the in vitro cultivation of animal circular ssDNA virus such as Porcine Circovirus 2 or human TT virus in an animal cell line. Increased titres of animal circular ssDNA virus are obtained by addition of interferons or agents which ensure the production of endogenous interferons by said cell line and / or by the reduction of endosomal-lysosomal system acidification.

The invention belongs to the field of biotechnology and microbial animal cell lines and particularly relates to a method for cultivating lymph-node autologous CIK (cytokine-induced killer) cells and application of the lymph-node autologous CIK cells. The CIK cells are obtained by in-vitro cultivation of isolated lymph node tissue obtained in surgery in patients with lung cancer and are compared with the CIK cells obtained from cultivation of isolated peripheral blood, biological characteristics of the CIK cells and cytotoxicity effect on pulmonary granule cancer cells are observed, the result shows that the isolated lymph node issue of the surgical patients can be completely used for cultivation of CIK, in-vitro tumor-cytotoxicity cells are powerful in activity, and the activity and cancer inhibition function are prior to the CIK cells of the peripheral blood. The CIK cells can be used for preparing pulmonary granule and cancer inhibition products and made into compounds with chemotherapy drugs as well as used for adoptive immunity and intervention for postoperative patients with lung cancers, and occurrence rate for recurrence and metastasis of the postoperative patients with the lung cancers is beneficially decreased.

The invention relates to a construction method and applications of engineering strain for delivering mammalian cell protein. The construction method comprises the following steps: deleting four virulence factors including exoS, exoT, exoY and ndk genes from pseudomonas aeruginosa PAK strain genome, so as to have no cytotoxicity to mammalian cells; further deleting popN gene, for inhibiting bacterial III type secretory system (T3SS), from chromosome, so as to remarkably increase the injection quantity of T3SS-mediated protein, thus constructing the engineering strain delta5. The constructed engineering strain delta5 can be used for delivering proteins of various mammalian cell linings, performing gene editing, cell reprogramming, protein function research and other work. The types of delivered cells are wide, including human and mice skin cells, muscle cells, intestinal cells, liver cells, multipotent stem cells and the like.

The present invention relates to the use of interferon in the in vitro cultivation of animal circular ssDNA virus such as Porcine Circovirus 2 or human TT virus in an animal cell line. Increased titres of animal circular ssDNA virus are obtained by addition of interferons or agents which ensure the production of endogenous interferons by said cell line and / or by the reduction of endosomal-lysosomal system acidification.

The invention relates to a method for detecting medicine adverse reaction with a genome expression profiling, which comprises the following steps: inquiring and confirming known medicines that can trigger specifical adverse reaction, namely risk medicines; counting the frequency of the adverse reaction of the risk medicines; utilizing the risk medicines and target medicines to respectively process human or animal cell lines, and obtaining the whole genome expression profiling of the cell lines through a genome expression chip technical platform; and counting the similarity of the target medicines and the corresponding risk medicines on the genome expression profiling, and carrying out weighting calculation for the similarity of the expression profiling according to the relative frequency of the adverse reaction of the risk medicines so as to score and judge the risk of the target medicine triggering the adverse reaction. The detection method is suitable for all kinds of medicines, without limitation, is simple and easy to operate, can indirectly estimate the risk of the target medicine triggering the adverse reaction without at the cost of practical personal injury, and has excellent application prospect for clinical medicine risk assessment.

The present invention relates to cancer cell system. The establishment of tumor metastasis animal model is essential for the deep research of tumor invasion and metastasis mechanism. SCID mouse is used as experiment target, and the establishment includes subcutaneously inoculating breast cancer parent MCF-7 cell suspension in shoulder of SCID mouse, killing SCID mouse 68 days after inoculation to take out lung for primary culture, cell passage after cells fill the bottom of the culture bottle with the cell being named as LM-MCF-7, subcutaneously re-injecting tumor cell to SCID mouse after 20 generation culture in vivo of LM-MCF-7 and repeating the said steps. The process has tumor forming rate of 100 % and wide metastasis in lung, kidney, spleen, marrow, lymph node, heart, etc. of SCIDmouse is found.

Modified cell lines and methods for use in the production of cultured meat are disclosed. The inventive methods provide for insertion of cell cycle regulatory genes or genes that encode for animal myoglobin into the genome of an animal cell to proliferate and flavor cell productions, followed by excising the inserted genes to terminate proliferation.

The invention provides a Ming human infiltrating type gastric cancer cell line and an application thereof and relates to a microorganism animal cell line. The gastric cancer cell line is a Ming human infiltrating type gastric cancer cell line XGC-1 with the preservation number of CCTCC NO:C201385. The invention also provides an application of the Ming human infiltrating type gastric cancer cell line XGC-1 in establishment of a cell model showing occurrence, development or transfer of a Ming infiltrating type gastric cancer, an application of the Ming human infiltrating type gastric cancer cell line XGC-1 in establishment of a Ming infiltrating type gastric cancer animal model, an application of the Ming human infiltrating type gastric cancer cell line XGC-1 in establishment of a cell model for studying a differentiation mechanism, cellular morphology, dysfunction and a tumor infiltration transferring mechanism of gastric cancer and guiding clinical comprehensive diagnosis and treatment in a Ming parting aspect and an application of the Ming human infiltrating type gastric cancer cell line XGC-1 in study of a gastric cancer occurrence mechanism and screening of medicines used for preventing and treating gastric cancer.

The invention provides a Ming classification human expansive type gastric cancer cell line and application thereof, and relates to a microorganism animal cell line. The Ming classification human expansive type gastric cancer cell line is a Ming classification human expansive type gastric cancer cell strain XGC-2 with the preservation number of CCTCC (China Center for Type Culture Collection) NO:C201386. The Ming classification human expansive type gastric cancer cell strain XGC-2 can be applied in a cell model of occurrence, development or transfer of Ming classification expansive type gastric cancer, applied to establishment of a Ming classification expansive type gastric cancer animal model, applied to a cell model for researching the differentiation mechanism, cellular shape and function abnormality, tumor invasion and metastasis mechanism and clinical comprehensive diagnosis guidance and the like from the aspect of Ming classification types, and applied to research of gastric cancer occurrence mechanism and screening of gastric cancer preventing and treating medicaments.

The invention belongs to the field of microorganism animal cell lines, and relates to a cell system for generating hepatitis B virus (HBV) recombinant cccDNA. Through a transposon system, HBV single-copy genomes with two ends provided with loxP sites are integrated into HepG2 cells, a clone HepG2-HBV / loxP cell line integrated with the different-copy-number HBV genomes is obtained, the cell line does not express HBsAg, after cyclization recombination enzyme (Cre) is introduced through adenovirus transduction, the rcccDNA is generated and the HBsAg is expressed, and expression of the HBsAg is closely relevant to the generation and level of the rcccDNA; the rcccDNA is rapidly and stably generated, the structure feature of the rcccDNA is similar to that of real HBV cccDNA, and transcription, copy and protein expression of viruses can be supported; the rcccDNA can be quantitatively detected through quantitative PCR by designing a specific primer, and Southern Blot detection can be carried out by utilizing a digoxin marking system. The cell system can be used for establishing a platform for HBV cccDNA relevant biological research and screening and evaluation of anti-HBV medicines.

The invention discloses a human intrahepatic bile duct cancer cell line ICC-X3 and application thereof, and belongs to the field of microbial animal cell lines. The provided intrahepatic cholangiocarcinoma cell line is a human intrahepatic cholangiocarcinoma cell line ICC-X3, and has been preserved in the China Center for Type Culture Collection on February 22, 2022, and the preservation number is CCTCC NO: C202260. The human intrahepatic cholangiocarcinoma cell line ICC-X3 can be used as a cell model for researching the occurrence, development or metastasis mechanism of intrahepatic cholangiocarcinoma. The human intrahepatic cholangiocarcinoma cell line ICC-X3 can also be used for establishing an intrahepatic cholangiocarcinoma animal model. The human intrahepatic cholangiocarcinoma cell line ICC-X3 can be used as a cell model for studying the differentiation mechanism, cell morphology and dysfunction and tumor infiltration and metastasis mechanism of intrahepatic cholangiocarcinoma, guiding clinical comprehensive diagnosis and treatment and the like.

The invention belongs to the field of animal cell line, and specifically relates to a human liver cancer cell line for observing life period of hepatitis B virus in cells and applications thereof. The human liver cancer cell line is characterized in that the human liver cancer cell line can secrete hepatitis B virus particles, and different loci of core antigens of hepatitis B virus particles all carry a TC label. The human liver cancer cell line can directly combine with bi-arsenic dye, so the processes that hepatitis B virus enters a cell, is transported, duplicated and expressed in the cell, and finally is transported out of the cell can be continuously and stably observed, and thus the characteristics of hepatitis B virus can be more visually observed.

The invention discloses a new application of protein Arginine methyltransferase 5. The new application provided by the invention is that protein Arginine methyltransferase 5 and the coding gene thereof are utilized to promote mammalian cells to express foreign proteins. The invention protects a method for preparing a recombinant mammalian cell line expressing foreign proteins. The method is as follows: the coding gene of Arginine methyltransferase 5 is introduced in the mammalian cells to obtain the recombinant mammalian cell line expressing foreign proteins. The invention also protects the recombinant mammalian cell line expressing foreign proteins prepared by the method. The recombinant cell line prepared by the method can be utilized to efficiently express the drug protein modified by glycosylation, the cell line has higer efficiency and lower demand on the purification process; and when applied in the production of bio-engineering drugs, the cell line has obvious economic benefit.

The invention belongs to the field of microbial and animal cell lines and provides an MDS (myelodysplastic syndrome) transfection leukocyte line with capacity of stable expression of GFP (green fluorescent protein) as well as establishment method and an application of the MDS transfection leukocyte line. A human MDS transfection leukocyte line SKM-1 is taken as maternal cells and is transfected through lentiviruses carrying GFP genes, GFP-positive single cells are obtained with a limiting dilution method and cloned, cells after amplified culture are screened in a mouse body in a subcutaneous injection manner, tumor masses are separated after tumorigenesis and cultured continuously, and the MDS transfection leukocyte line SKM-1 / GFP with stable expression of GFP is obtained. The morphology, growth characteristics and the growth curve of the cell line have no difference with those of the maternal cells, and the cell line can express GFP stably, has tumorigenicity, can be further applied to establishment of a murine animal model and provides a platform for MDS, minimal residual disease and other preclinical study.

The present invention relates to a recombinant expression vector for an animal cell containing a dihydrofolate reductase (DHFR) coding nucleotide sequence operatively linked to a DHFR promoter, to an animal cell line transformed by the vector, and to a method for preparing a target protein using the same. As compared with existing animal cell expression vectors, the vector of the present invention enables an effective screening of a cell line clone in which foreign genes are amplified together with DHFR genes even at a much lower methotrexate concentration. The present invention exhibits excellent effects in cell line preparation as high-productivity cell lines can be ensured in a short time through the use of a lower concentration of methotrexate in the process of protein production cell line establishment.

The invention belongs to the filed of microorganism animal cell lines and provides a Chinese lung adenocarcinoma cell line with high metastases potentiality of bone, lever and salivary gland. According to the invention, hydrothorax of a lung adenocarcinoma first-diagnosis patient is used as primary cells for culturing; the human lung adenocarcinoma cell has the preservation number of CGMCC No.3138 and classified name as human lung adenocarcinoma cell line CPA-Yang4, with the tumorigenesis rate up to 50-100 percent; and the human lung adenocarcinoma cell can grow quickly and metabolize vigorously and has the biological characteristics of mainly transferring the bone, transferring the lever and the salivary gland and other organs as well as classic pathological characteristics of transferring tumor CPA-Yang4 bone and salivary gland. The invention can be used for providing reference data to early diagnosis for transferring the human lung adenocarcinoma bone and further analyzing lung cancer transferred relevant functional genomes and transferred relevant genes and can provide an effective molecule detection method for early diagnosis and curative effect assessment for bone transfer.

The invention discloses a multifunctional gene expression platform regulated by green tea metabolite protocatechuic acid. The platform relates to an on-off system regulated by protocatechuic acid, i.e. the expression is initiated or closed by the protocatechuic acid regulation gene. The on-off system comprises a protocatechuic acid regulated 'on' system and a protocatechuic acid regulated 'off' system. The invention also provides a eukaryotic expression vector comprising the on-off system, a mammalian cell line and a microcapsule. The invention also discloses an application of the multifunctional platform capable of the accurate expression release of controllable insulin or glucagon-like peptide in treating diabetes; and the gene expression regulation platform can be used for establishinga complicated biological computer as well as regulating the gene edition and gene expression activation or inhibition mediated by a CRISPR / Cas9 or CRISPR / dCas9 system. The multifunctional gene expression platform provides a useful novel gene expression regulation tool for the gene treatment and cell treatment.

The present invention belongs to the field of microbial animal cell lines, and specifically relates to a human diffuse large B-cell lymphoma glucocorticoid-resistant cell line Toledo / DEX having a glucocorticoid-resistant potential. The GR [alpha] expression level of the Toledo / DEX cell strain of the invention is below source cells, and the drug-resistant multiple of the cell strain for dexamethasone is higher than the source cells. The source cells are the diffuse large B-cell lymphoma cell line. The cell line is the first case of the human diffuse large B-cell lymphoma glucocorticoid-resistant cell line having the glucocorticoid-resistant potential at home and abroad. The cell line shows that establishing lymphoma drug-resistant cell lines by a long-term, intermittent, low-dose glucocorticoid increment induction method is practically feasible. The establishment of the Toledo / DEX cell line provides an experimental platform for subsequent glucocorticoid drug-resistant mechanisms and reversal.

The invention belongs to the field of microorganism animal cell lines, and especially relates to a pair of human stomach cancer cell lines having high metastasis towards livers and lungs and an establishing method thereof. The human stomach cancer cell lines are characterized in that the human stomach cancer cell lines are MKN28-M and SGC7901-M respectively; the MKN28-M is assigned the accession number CGMCC No.6901; and the SGC7901-M is assigned the accession number CGMCC No.6902. The cell lines are humanized and have no obvious difference from parent cells in in-vitro growth and multiplication capacities, but have obviously enhanced migration, invasion and transfer capabilities over the parent cells. Results of nude mouse tail vein injection visceral metastasis experiments show that the average rate of metastasis towards livers and lungs is 70%. The cell lines have characteristics of high metastasis capacity and high malignancy degree. An animal model established based on the cell lines provides a good platform for further mechanism study of human stomach cancer metastasis. The animal model is an effective and feasible experiment model used for studying mechanisms of human stomach cancer metastasis and evaluating anti-cancer drug curative effects at present.