53 results about "Established cell line" patented technology

The invention relates to a method for establishing and identifying a cynoglossus semilaevis testis cell line, which comprises the following steps: adopting a MEM (Minimum Essential Medium), adding 20 percent of fetal calf serum, 0.1 percent of B-mercaptoethanol, 2 ng / ml of human alkaline fibroblastic growth factors, 1 ng / ml of leukemia inhibiting factors, 1 nmol of sodium pyrurate and 1 nmol of glutamine and preparing into a complete medium; inoculating a tissue block into a culture bottle, overturning and inversely placing the bottle to culture for 3 h and positively placing the culture bottle after the tissue block is attached onto a wall, so that the tissue block is soaked into the culture medium for culture. A trypsin digestion method is adopted for subculturing cells. One cynoglossus semilaevis testis cell line is established by adopting the above method and is subcultured to fiftieth generations. Meanwhile, the invention also provides a method for identifying a chromosome of the established cell line and the level of a molecular marker and a method for identifying the sensitivity of the cell line to related viruses. The method for establishing the cynoglossus semilaevis testis cell line can be used for culturing other fish gland cells and establishing the cell line, thereby having wider promotion and application prospect.

The invention discloses a Pseudosciaena crocea muscle cell line and its establishing method. The cell line is preserved in China Center for Type Culture Collection on Oct., 31, 2013, and has a preservation registration number of CCTCC NO:C2013158. The Pseudosciaena crocea muscle cell line can be serially passed, can provide a large amount of the Pseudosciaena crocea muscle cell line, and can be used for researching pathogenic characteristics, vaccines, gene functions and breeding; and the Pseudosciaena crocea muscle cell line has excellent properties, is a fibroblast and is soma-fusiform or anomaly triangular, the cytoplasm outwardly stretches to form a plurality of protrusions with different lengths, and the protrusions are arranged in a radial or spiral manner. The Pseudosciaena crocea muscle cell line has the characteristics of strong splitting, short passage time, easy digestion, good adherent rate and starvation resistance. The establishing method has strong repeatability, and the established cell line has a good stability.

The invention aims at providing an establishment method of an ovary cell line of cynoglossus semilaevis, which comprises the following steps: 1) removing the outermost membrane of the ovary tissue of cynoglossus semilaevis under a sterile condition, cutting the tissue into small blocks in a culture solution, flushing the tissue blocks with a PBS solution and digesting with a trypsin solution, wherein the solution after trypsin digestion contains cells, cell clusters and tissue blocks not completely digested; and 2) centrifuging the solution to remove the supernatant trypsin, and suspending the cells, cell clusters and small tissue block precipitates with a culture solution; after a suspension is obtained, inoculating into a culture plate treated by the type-I collagen; adding a culture solution, and culturing in a biochemical incubator at 24 DEG C; and performing subculture to finish establishment of the ovary cell line. In the establishment method provided by the invention, an ovary cell line of cynoglossus semilaevis is successfully established by use of the ovary tissue of cynoglossus semilaevis, the established cell line can be continuously passed, the passage cells obtained by the method can be passed for over 60 generations, and a great quantity of ovary cells can be provided.

An object of the present invention is to provide a cytotoxic assay using a new established fish cell line. The invention attained according to the object is a cytotoxic assay, wherein an evaluation of toxicity of a specimen is performed on the basis of its toxicity to an established cell line originated from sturgeon, preferably a STIP-1 cell line (FERM BP-8421) and a STIP-3 cell line (FERM BP-8422). Furthermore, another object of the present invention is to provide a new established cell line expected to be used in the diagnosis of viral infection disease of sturgeon and so on. The invention attained according to the object is an established cell line originated from a sturgeon eyeball, particularly a STIP-1 cell line (FERM BP-8421) and a STIP-3 cell line (FERM BP-8422).

The invention relates to mouse stem cell culture technologies, and specifically discloses a construction method of mouse bone marrow mesenchymal stem cell lines. The construction method of the mouse bone marrow mesenchymal stem cell lines comprises the following steps: extracting bone marrow; carrying out primary culture of mesenchymal stem cells; carrying out subculture; carrying out cryopreservation; carrying out resuscitation; carrying out molecular identification of cell surface; and identifying differentiation into adipocytes and osteoblasts. Cell culture fluid adopted for the construction method of the mouse bone marrow mesenchymal stem cell lines is a low-sugar-content DMEM basic cell culture medium added with fetal bovine serum. The constructed mouse bone marrow mesenchymal stem cell lines are fibroblast-like in morphology. The construction method of the mouse bone marrow mesenchymal stem cell lines disclosed by the invention is simple in operation and high in repeatability; and the constructed mouse bone marrow mesenchymal stem cell lines are feasible for continuous passage, thereby enabling supply of a large number of mesenchymal stem cells. In addition, the constructed mouse bone marrow mesenchymal stem cell lines can differentiate into adipocytes and osteoblasts, and can be directly applied in study on functional genes of mice. The construction method of the mouse bone marrow mesenchymal stem cell lines is expected to become a research platform of stem cell functions and regulation mechanisms, and is capable of promoting practical application of tissue engineering on basis of mesenchymal stem cells.

The present invention relates to a method for picking a colony of cells and to producing a cell line from this colony, as well as to the cell line thus established and to use of this cell line in a method for producing a biological molecule or biological entity of interest.

An object of the present invention is to provide a cytotoxic assay using a new established fish cell line. The invention attained according to the object is a cytotoxic assay, wherein an evaluation of toxicity of a specimen is performed on the basis of its toxicity to an established cell line originated from sturgeon, preferably a STIP-1 cell line (FERM BP-8421) and a STIP-3 cell line (FERM BP-8422). Furthermore, another object of the present invention is to provide a new established cell line expected to be used in the diagnosis of viral infection disease of sturgeon and so on. The invention attained according to the object is an established cell line originated from a sturgeon eyeball, particularly a STIP-1 cell line (FERM BP-8421) and a STIP-3 cell line (FERM BP-8422).

The present invention belongs to the field of biotechnology and cell engineering, and relates to monoclonal antibody hybridoma cell line and its construction process. Technology, the present invention features that soluble immunogen is first prepared with human ovary carcinoma tissue sample and then used to immunize BALB / c mouse, and through fusion, screening and repeated cloning in hybridoma technology, hybridoma cell line capable of secreting stably human ovary carcinoma resisting monoclonal antibody is obtained. The hybridoma cell line, named as BUPH: OVCAMab, is preserved in the preservation number of CGMCC No. 1225. The present invention also discloses the method of preparing monoclonal antibody OVCAMab with the cell line and purifying the antibody. The antibody is used in early diagnosis, intracorporeal locating, detection, metastatic focus detecting, etc. of ovary carcinoma in radioimmunoassay.

The present invention relates to a method for screening a compound for preventing, ameliorating or treating glomerular lesions or disease of kidney, characterized by assaying the regulating activity of compound on 4F2hc expression shown by contacting human peripheral blood mononuclear cells or monocytes or an established line of human cultured cells having a nature of human macrophages with macrophage-activating substances. The screening method is useful for screening compounds capable of preventing, ameliorating or treating glomerular lesions or disease of kidney.

The present invention relates to Lymantria xylina cell lines established from pupal tissues of L. xylina Swinhoe, including NTU-LY-1, NTU-LY-2, NTU-LY-3, and NTU-LY-4. These four cell lines were confirmed to be newly established cell lines derived from L. xylina by random amplified polymorphic DNA-polymerase chain reaction (RAPD-PCR) and isozyme analysis. The genotypes and characteristics of the abovementioned cell lines are totally different from other insect cell lines. In addition, these four L. xylina cell lines are susceptible to insect baculovirus of L. xylina multiple nucleopolyhedrovirus (LyxyMNPV), LdMNPV-like virus, PenuMNPV, as well as microsporidia of Nosema sp. (isolated from Eurema blanda) and N. bombycis and the like. Therefore the invention can be applied in multiplication of the abovementioned insect-pathogenic microorganisms to produce biopesticides in pest control. In addition, the cell lines can also be used as hosts for the expression vectors of said baculovirus to produce recombinant proteins.

Belonging to the technical field of genetic engineering, the invention relates to a SgRNA targeting sequence for knockout of mouse Tet3 gene, a cell line for conditional induced knockout of mouse spermatogonia Tet3 gene, a construction method and application thereof. The gene editing off-target effect is an important aspect that puzzles the application of gene editing technology. The Tet3 gene target screened by the patent is the only sequence-targeted target screened out from the whole mouse genome, no similar sequence exists in the genome, and therefore off-target effect does not exist theoretically. At the same time, an induced knockout method is employed to close Cas9 protein expression and also can effectively prevent off-target, 24h later after adding of Dox, 97% or more of the mousespermatogonia genome can be knocked out, and the cell lines before and after Dox induction can be employed as good experimental group and control group. The cell line and method established by the invention can be widely applied to Tet3 and other genetic functional study.

The present invention provides a method for testing an allergy capable of rapidly and highly accurately testing an allergic reaction. The method can determine whether or not a patient has an allergy or whether or not an agent that may be allergenic to a patient has an allergenicity (an allergic reactivity) in the patient. The method may comprise the steps of causing migration of leukocytes separated from a healthy human or cells of an established cell line with a chemotactic factor contained in a sample such as body fluid or blood of the patient to be tested or a sample stimulated with the agent that may be allergenic to the patient and analyzing the cell kinetics such as migration velocity, migration distance, and migration direction.