9 results about "Fibroblast cell line" patented technology

This invention relates to the interdisciplinary field of cell biology, experimental oncology, and translational medicine, specifically to a cancer-associated fibroblast cell line, its construction method, and its applications. This invention fills the research gap in mouse-derived oral squamous cell carcinoma-associated fibroblast cell line models and provides a method for constructing such a cell line. Through the inverted adherent culture and enzymatic digestion methods of this invention, a mouse-derived oral squamous cell carcinoma-associated fibroblast cell line with a clear origin, high purity, and long-term phenotypic and functional stability can be obtained. This allows for the establishment of a cryopreservable and resuscitable mouse oral squamous cell carcinoma-associated fibroblast cell bank, providing technical support for research related to the tumor microenvironment and immunotherapy.

The invention discloses a rumen fibroblast line as well as a construction method, a culture method and application thereof, the rumen fibroblast line G8 is preserved in the China Center for Type Culture Collection, the preservation number is CCTCC NO: C2025332, the rumen fibroblast line G8 has a typical fibroblast-like form and can be stably passaged, and the rumen fibroblast line G8 can be used as a rumen fibroblast cell line. The cell culture medium also has the characteristic of highly expressing fibroblast marker genes and the function of tolerating and utilizing rumen core metabolite butyric acid, and the proliferation rate and metabolic activity of the cell culture medium under the condition that 15mmol / L butyric acid is used as a main energy source are obviously superior to those of glucose culture; according to the construction method of the rumen fibroblast line, the steps of function screening and molecular identification are introduced, so that the identity authenticity of the constructed cell line can be ensured; the culture method of the rumen fibroblast line is simple and effective, culture medium materials are easy to obtain, and culture conditions are easy to realize; the rumen fibroblast line can be used for rumen in-vitro research, and is beneficial to clarification of a mechanism for regulating rumen interstitial metabolism and tissue repair by butyric acid.

An sgRNA targeting a porcine PLIN1 gene and use thereof. The sgRNA action site is located on the second exon and the fourth exon of the porcine PLIN1 gene. Provided is a method for knocking out a PLIN1 gene in a porcine fetal fibroblast cell line by using a CRISPR / CAS9 system. The method comprises designing and synthesizing an sgRNA targeting a porcine PLIN1 gene according to a porcine PLIN1 gene sequence, then constructing a CRISPR / Cas9 targeting vector containing the sgRNA, and transferring same into a porcine fetal fibroblast cell to obtain a PLIN1 gene-knocked-out porcine fetal fibroblast cell. The cell can be used for the preparation of a PLIN1 gene-edited porcine, and is not only of great significance for porcine breeding, but also of great reference value for revealing the occurrence mechanism of human hereditary lipodystrophy and seeking a treatment method.

The invention provides a sheep embryo fibroblast line and application thereof, and belongs to the technical field of cell biology. The name of the sheep embryo fibroblast line is sheep embryo fibroblast OEF, the sheep embryo fibroblast line is preserved in China Center for Type Culture Collection, the address is Wuhan University, Wuhan, China, the preservation date is January 26, 2022, and the preservation number is CCTCC NO: C202212. The sheep embryo fibroblast line can be used for separation and culture of bovine and sheep pox viridae viruses such as LSDV, GTPV, SPPV and ORFV, has the characteristics of short proliferation time and high virus titer, and can avoid the risk that other viruses are polluted by viruses generated by primary cell separation.

The invention relates to the technical field of molecular biology and genetic engineering, and discloses sgRNA targeting a chicken CD30 gene and a combination and application thereof. The sgRNA and the sgRNA combination provided by the invention perform targeted knockout on the seventh exon region (transmembrane region) of the chicken CD30 gene, and the sgRNA knockout target of the chicken CD30 is transfected into a chicken fibroblast line DF-1 to prepare the CD30 knockout DF-1 cell, namely the CD30 / -DF-1 cell, and compared with a control group cell through detection, the CD30 expression quantity in the CD30 / -DF-1 cell is extremely low, and the result shows that the CD30 is successfully edited. The establishment of the CD30- / -DF-1 cell line provides an experimental material for deeply researching the functions of CD30.

This invention belongs to the field of animal cell culture technology, specifically relating to chicken embryo fibroblast cell lines, their culture media, acclimatization methods, and applications. This application provides a chicken embryo fibroblast cell line derived from… Gallus gallus The chicken embryo fibroblast suspension cell line CEF-P17 3D is deposited at the China Center for Type Culture Collection (CCTCC) with accession number CCTCC NO: C202661 and deposit date of March 31, 2026. This application also provides a culture medium for culturing the chicken embryo fibroblast cell line and a method for acclimatizing the chicken embryo fibroblast cell line, which shortens the acclimatization period, simplifies the operation, and is suitable for large-scale culture. This application also provides the application of the culture medium for the chicken embryo fibroblast cell line. Furthermore, this application also provides the application of the chicken embryo fibroblast cell line in the preparation of cultured meat.

The invention provides a preparation method and application of an in-vivo CAR-T cell for treating interstitial lung disease, and provides a T cell of a chimeric antigen receptor which is modified by genetic engineering and is used for expressing targeted fibroblast activating protein. A targeted fibroblast activation protein (FAP) CAR-T cell therapy model is constructed by targeting CD5 entrapped mRNA nano-liposome (LNP) transfection and lentiviral vector, and the method comprises the following steps: firstly, verifying the difference of the killing ability of FAP CAR-T constructed by CD5 LNP-mRNA and lentivirus in a 293T cell which stably co-expresses FAP, Luciferase and mCherry, and then verifying the difference of the killing ability of FAP CAR-T constructed by LNP-mRNA and lentivirus in the 293T cell which stably co-expresses FAP, Luciferase and mCherry; further verifying the effectiveness of the CAR-T cells constructed by transfecting the CD5 LNP-mRNA on cell lines of human fibroblasts (CDD19Lu, LL29 and LL97A) and primary fibroblasts of human and mice in vitro, and further verifying the effectiveness and safety of the CD5 LNP-FAP CAR-T in treatment of pulmonary fibrosis through in-vivo experiments in animals. A novel and effective anti-fibrosis treatment thought is provided for patients with fibrosis interstitial lung diseases.

This application provides a method for establishing and applying mouse gastric squamous cell carcinoma cell lines and gastric cancer-related fibroblast cell lines. The culture medium for culturing these cell lines comprises L-WRN cell culture supernatant, and 100×N-2 additive (0.5 vol%–1.5 vol%), 50× vitamin A-free B-27 additive (1.5 vol%–2.5 vol%), N-acetylcysteine ​​amide (1 mM–1.5 mM), nicotinamide (8 mM–12 mM), FGF-10 (80 ng / mL–120 ng / mL), EGF (40 ng / mL–60 ng / mL), A83-01 (0.8 μM–1.2 μM), Y-27632 dihydrochloride (8 μM–12 μM), Glutamax additive (0.5 vol%–1.5 vol%), and Matrigel. The culture medium provided in this application contains 5 vol%–7 vol% Hepes buffer (8 mM–12 mM) and gastrin (0.8 nM–1.2 nM). This medium can significantly improve the success rate of in vitro culture of primary cell lines. The mouse gastric squamous cell carcinoma cell lines and gastric cancer-related fibroblast cell lines obtained from this medium are genetically stable and can be used to construct ectopic or orthotopic implantation models in immune-active mice.

The present application relates to a method for constructing a pig endogenous retrovirus inactivated xenotransplant donor pig, and belongs to the technical field of xenogenic organ transplantation. The copy number and PERV-pol genotype of PERVs are determined by ddPCR, Sanger sequencing and second-generation sequencing. Based on the RNP system of CRISPR / Cas9, the pig PERV-pol gene targeting sequence is designed and synthesized, and is co-transfected into a pig fetal fibroblast cell line with spCas9 protein, positive cells are screened, somatic cell cloning and embryo transfer are carried out, and the PERV-pol knockout condition of the fetus is identified when pregnancy occurs. If not completely knocked out, the sgRNA targeting sequence needs to be designed and synthesized again, and the cell line with completely knocked out PERV-pol gene is screened. If completely knocked out, somatic cell cloning and embryo transfer are directly carried out; and the xenotransplant donor pig with inactivated PERVs is obtained.