9 results about "3T3 cells" patented technology

A recombinant humanized collagen type III alpha-1 (rhCol III a1), and an expression vector and use thereof are provided, relating to the technical field of bioengineering. A sequence on positions 154 to 1,221 of a human type III collagen α chain is ligated to an optimized vector pcDNA3.1, a resulting optimized expression vector is transfected into a human embryonic kidney cell Expi293F, and then rhCol III a1 is secreted to form a fusion protein with a soluble green fluorescent protein (GFP). The expressed rhCol III a1 that forms the fusion protein with the GFP shows an activity of promoting the migration of BALB / c 3T3 cells, thus exhibiting a great application potential in the field of skin care.

The invention provides a pair of sgRNAs targeting a mouse FAM50A gene and application of the sgRNAs, and belongs to the technical field of gene editing, the sgRNAs comprise sgRNA1 and sgRNA2, recognition sites of the sgRNA1 and the sgRNA2 are located in the third exon region and the fourth exon region of the mouse FAM50A gene respectively, the sequence of the sgRNA1 is as shown in SEQ ID NO.1, and the sequence of the sgRNA2 is as shown in SEQ ID NO.2. Based on the sgRNA1 and sgRNA2 of the targeted FAM50A gene provided by the invention, an NIH / 3T3 cell line and a beta-TC-6 cell line of which the FAM50A gene is stably knocked out can be efficiently constructed by combining a CRISPR / Cas9 system, and a research tool and a material are provided for the functional research of the FAM50A gene and the development of related disease models.

The invention discloses a polypeptide for promoting collagen synthesis and application thereof. The polypeptide capable of promoting collagen synthesis is specifically combined with a receptor TbetaR II to regulate collagen synthesis in fibroblasts, and is a TGF-beta mimic peptide analogue. And the TB2 has a proliferation effect on BALB / 3T3 cells. The TB2 can promote the expression of collagen related factors Collagen I, Collagen III, Smad3 and MAPK, and has the effects of promoting cell migration and promoting wound healing. Therefore, the polypeptide for promoting collagen synthesis provided by the invention can be used for preparing a preparation for promoting cell proliferation or collagen generation.

This application provides a bone-targeting biomimetic biomineral nanocomposite material, its preparation method, and its application, relating to the field of medical materials. The method includes the following steps: S1: Providing water-soluble phosphate and water-soluble calcium salt; S2: Mixing the aqueous solutions of the water-soluble phosphate and water-soluble calcium salt, followed by magnetic stirring and centrifugation to obtain calcium phosphate; S3: Washing the calcium phosphate and freeze-drying for storage; S4: Providing GLG1+ NIH-3T3 cells and expanding them; S5: Extracting the cell membrane from the GLG1+ cells and resuspending the cell membrane in pre-cooled PBS buffer to obtain a cell membrane solution, which is stored at -80°C for later use; S6: Mixing the calcium phosphate from S1 with the GLG1+ cell membrane solution and sonicating to obtain cell membrane-coated calcium phosphate nanoparticles; S7: Centrifuging the calcium phosphate nanoparticles to remove excess cell membrane, resuspending them in ultrapure water and washing at least twice to finally obtain the bone-targeting biomimetic biomineral nanocomposite material, which is then stored at 4°C.

The application discloses a high-throughput anti-fibrosis drug screening method based on a fluorescence-free image and application thereof, relates to the technical field of biological medicine, and aims to obtain a rapid and simple method for large-scale drug screening. In the application, DPC images of NIH / 3T3 cells are collected by a high-content instrument, correlation analysis is performed on morphological parameters obtained by self-analysis software and the expression amount of alpha-SMA protein, 13 morphological parameters highly correlated with the expression amount of alpha-SMA protein are screened out, and the 13 morphological parameters are combined with an artificial intelligence method to be used for large-scale drug screening. The method has the advantages of high throughput, high accuracy, simple operation, and the like, can be used to determine whether a drug has an anti-fibrosis effect, can improve detection efficiency, can be applied to prediction of whether a traditional Chinese medicine monomer has an anti-fibrosis effect, and proves the accuracy and effectiveness of the method as an anti-fibrosis drug screening method, thereby providing important experimental basis and technical support for future drug development and treatment of fibrosis diseases.

This disclosure provides a PD-1 antibody and its application. The PD-1 antibody or antigen-binding moiety provided in this disclosure has a high affinity for human PD-1 protein and can bind at a rate of 0.9 x 10⁻⁶. ‑8 The Kd value of M binds to human PD-1. Furthermore, the binding specificity is strong, and it does not cross-react with human CTLA-4 positive cells (3T3 cells overexpressing human CTLA-4) or human PD-1 negative cells (such as Raji cells). Based on these characteristics, the PD-1 antibody or antigen-binding moiety provided in this disclosure can be used for the detection of human PD-1 protein, and can also be used alone or in combination with other methods in tumor immunotherapy. That is, it can be effectively applied in the preparation of drugs for treating tumors, infectious diseases, autoimmune diseases, and anti-immune rejection.

The invention provides a PD-1 antibody and application thereof. The PD-1 antibody or antigen binding part provided by the invention has high affinity with human PD-1 protein, and can be bound with human PD-1 at a Kd value of 0.9 * 10 <-8 > M. Moreover, the binding specificity is high, and cross reaction with human CTLA-4 positive cells (3T3 cells overexpressing human CTLA-4) and human PD-1 negative cells (such as Raji cells) is avoided. Based on the characteristics, the PD-1 antibody or the antigen binding part provided by the invention can be used for detecting human PD-1 protein, and can also be independently applied to tumor immunotherapy or applied to tumor immunotherapy in combination with other methods, namely, the PD-1 antibody or the antigen binding part can be effectively applied to preparation of medicines for treating tumors, infectious diseases, autoimmune diseases, immunological rejection and the like.

A recombinant humanized collagen type III alpha-1 (rhCol III α1), and an expression vector and use thereof are provided, relating to the technical field of bioengineering. A sequence on positions 154 to 1,221 of a human type III collagen α chain is ligated to an optimized vector pcDNA3.1, a resulting optimized expression vector is transfected into a human embryonic kidney cell Expi293F, and then rhCol III α1 is secreted to form a fusion protein with a soluble green fluorescent protein (GFP). The expressed rhCol III α1 that forms the fusion protein with the GFP shows an activity of promoting the migration of BALB / c 3T3 cells, thus exhibiting a great application potential in the field of skin care.

This invention provides an application of a drug to inhibit silicosis fibrosis. This invention demonstrates that rhodioloside can inhibit the expression levels of fibrosis-related proteins Collagen 1 and α-SMA in SiO2-exposed mice, reduce the number of diffuse high-density shadows and characteristic silicotic nodules in the interstitial lung fields, and decrease the area of ​​diffuse collagen fiber deposition. Simultaneously, rhodioloside can also inhibit TGF-β-induced expression levels of Collagen 1 and α-SMA in NIH / 3T3 cells, inhibiting the transdifferentiation of fibroblasts into myofibroblasts. Furthermore, the research results show that the effect of the same dose of rhodioloside is comparable to, or even superior to, the current first-line clinical drug for alleviating silicosis fibrosis, tetrandrine. These research results prove that rhodioloside can effectively alleviate silicosis fibrosis.