56 results about "3T3 cells" patented technology

The invention discloses a target quaternary ammonium salt cationic polymer lipid gene carrier, a preparation method and an application thereof. The target quaternary ammonium salt cationic polymer lipid genetic carrier is characterized in that: a polymeric quaternary ammonium salt and lipid are adopted for preparing a quaternary ammonium salt cationic polymer lipid genetic carrier according to a mass ratio, wherein the mass ratio of the polymeric quaternary ammonium salt to the lipid is 0.05-20:1; then a assembly method or a modification method is adopted for modifying to prepare a folic acid or EGFR antibody modified cationic polymer lipid gene carrier. Results of gene transfection experiments show that: gene transfection efficiencies of the target quaternary ammonium salt cationic polymer lipid gene carrier in 293T cells and NIH-3T3 cells are the same as the gene transfection efficiencies of positive control lipofectamine of lipofectamine<TM>2000 in the 293T cells and the NIH-3T3 cells; the gene transfection efficiencies of the EGFR antibody modified cationic polymer lipid genetic carrier in liver cancer Huh-7 cells and breast cancer MCF-7 cells are higher than the gene transfection efficiencies of the lipofectamine<TM>2000 in the liver cancer Huh-7 cells and the breast cancer MCF-7 cells. The cationic polymer lipid genetic carrier system provided by the present invention has good biocompatibility and low cytotoxicity, and can be as an excellent non-viral gene delivery carrier.

The invention belongs to the technical field of zoobiology, and relates to an establishment method of a positive screening system based on gene knockout cells; on the basis of the established system,p53 and MAD2 genes are knocked out in NIH/3T3 cells, and the cells, which undergo gene knockout, are subjected to positive screening. According to the establishment method, target sequences of the sixth exon of the p53 gene and the first exon of the MAD2 gene are subjected to site-directed knockout in an NIH/3T3 gene line, and in the combination with a green fluorescent protein gene and a puromycin gene, the cells, which undergo p53 and MAD2 gene knockout, are subjected to positive screening; and by virtue of the established positive screening system, the p53 and MAD2 genes are simultaneouslyedited in living mouse liver, so that a mouse liver cancer model is finally obtained. The method provided by the invention lays a foundation for related researches on editing the p53 and MAD2 genes ina living mouse.

The invention belongs to the technical fields of medicinal chemistry and polypeptide biology, and relates to an oligopeptide derivative P15 capable of combination of basic fibroblast growth factor (bFGF), wherein the sequence of the P15 is N'-PILQAGLGGGS-NH2-C'. The invention specifically relates to the sequence of the P15, and an application of the P15 in fields of basic research and medicine. The present invention further comprises various peptide derivatives of the P15, wherein the P21 is the peptide derivative of the P15, and the sequence of the P21 is N'-PLLQA TA GGGS-NH2-C'. With combination of the bFGF, the P15 and the P21 can provide good inhibition activities for bFGF-induced NIH3T3 cell proliferation and bFGF-induced Bable/C 3T3 cell proliferation, can significantly inhibit proliferations of bFGF-induced lung cancer A549 cells and bFGF-induced glioma U251 cells, and provide prospects for development into anti-tumor peptide drugs.

The invention discloses a recombinant human fibronectin III1-C (rhFNIII1-C), and a preparation method and application thereof. The preparation method comprises the following steps: optimizing a targetgene segment of the target protein according to codon expression preference, and inserting the obtained gene segment into pET-32a to construct a recombinant plasmid; transforming and introducing therecombinant plasmid into an expression vector BL21(DE3), and carrying out screening to obtain positive clone bacteria capable of realizing efficient soluble expression; carrying out enlarging fermentation culture on the positive clone bacteria, carrying out induced expression, performing crushing and centrifuging to obtain a supernatant, and carrying out purification through elution with a His-tagaffinity chromatography column, dialysis, filtration and other steps to obtain a high-quality rhFNIII 1-C solution with a protein concentration of 1.0 mg/mL or above and a purity of 90% or above. According to detection and observation results of cell adhesion promoting tests, the rhFNIII1-C has the performance of obviously promoting adherence and adhesion of MDBK cells and Balb/c/3T3 cells and rapid division and growth of the cells, which indicates that the rhFNIII1-C has huge potential of being used as a raw material and a finished product for cosmeceuticals and medical skincare.

The invention provides a vitamin B2 modified IONzyme, and in particular to vitamin B2 modified nano-scale ferriferrous oxide particles. The invention further provides a preparation method of a vitaminB2 modified IONzyme and a composition for promoting oral ulcer healing. The vitamin B2 modified IONzyme provided by the invention has higher catalase activity and superoxide dismutase activities thanthe IONzyme; and compared with the individually used vitamin B2 and IONzyme, the vitamin B2 modified IONzyme presents better protection effect for BALB/3T3 cells and HOK cells in a hydrogen peroxideenvironment, better ROS clearance at a cellular level, and better ability to reduce IFN-gamma and IL-6 in ulcers. The vitamin B2 modified IONzyme has a better therapeutic effect on oral ulcers and canbe used to prepare effective drugs for promoting the healing of oral ulcers.

The invention relates to a Scid mouse model of electromagnetic radiation carcinogenesis and a constructing method thereof. The method comprises the following steps of: culturing an NIH/3T3 cell and carrying out passage; exposing the cell in the presence of the electromagnetic radiation of 30-90W/m<2> and culturing for 4-6 weeks; and carrying out a soft agarose culturing experiment and inoculating the cultured soft agaroes into a Scid mouse for carrying out an oncogeen experiment. The result shows that a cell colony is formed in the soft agarose experiment and Scid mice in the oncogeen experiments all generate tumors. By using the method, the Scid mouse model with tumors induced by using the electromagnetic radiation can be established; and the method has the advantages of remarkable effect, simple steps and favorable repeatability. An in vitro cell transformation experimental model established by the method can be used for simulating the vicious transformation of an in vivo cell and provides a favorable experimental model and a powerful technical means for the research on a pathogenesis and prevention and treatment measures of the tumor induced by the electromagnetic radiation. The method can be applied to the carcinogenic risk pre-evaluation on the electromagnetic radiation in the industry, science, medical treatment and daily life and has an important value of preventing the long-term harm of the electromagnetic radiation on the human health.

The invention provides a purification preparation method and an application of mesenchymal stem cell secreted factors. The purification preparation method comprises the following steps: (1) preparing MSC supernate; (2), purifying the mesenchymal stem cell secreted factors, including filtration and concentration, heparin affinity chromatography, and heparin affinity chromatography elution peak 5kD ultrafiltration or G-25 desalination column desalination; and (3) using the purified MSC secretion factors in the step (3) to prepare masks. According to the invention, a heparin affinity chromatography medium is adopted to specifically adsorb mesenchymal stem cell (MSC) secreted factors and purify substances with an effect of stimulating NIH 3T3 cell proliferation in MSC secretions, and the substances are potentially used for repairing refractory wounds and are added to cosmetics as an additive. The effects of repairing damaged skin, eliminating skin wrinkles, shrinking pores, improving complexion and the like can be achieved.

The invention belongs to the technical field of bioengineering, and particularly relates to EGF-like protein and a construction method thereof, and chimeric protein as well as a preparation method andapplication thereof. The invention provides the EGF-like protein; the EGF-like protein is EGF-CTA2-TAT protein; the nucleotide sequence of the EGF-CTA2-TAT protein is shown in SEQ ID NO: 1. The EGF-CTA2-TAT protein is chimeric with EGF protein with a repair effect, cell-penetrating peptide assisting transdermal absorption, and a cholera toxin A2 subunit (CTA2) with the ability of penetrating cellmembranes; it is shown by a cell activity experiment test result that the EGF-CTA2-TAT protein has obvious effect of promoting the growth of BALB/c 3T3 cell strains, and has bioactivity.

The invention provides a OsrbFGF gene optimized by paddy rice genetic codon, correlated vectors, a method for preparing OsrbFGF transgenic paddy rice seeds, and a separation purification method of OsrbFGF. The OsrbFGF transgenic paddy rice seed is prepared through specific expression of OsrbFGF by paddy rice endosperm cells, and OsrbFGF is separated and purified. The purity of obtained OsrbFGF is 95%, and obtained OsrbFGF has the activity of promoting NIH/3T3 cell proliferation in vitro and promoting wound healing in vivo.

The invention relates to application of a chicken anemia virus (CAV)-derived VP1-aa 23-43 polypeptide as an efficient cell penetrating peptide. The sequence of the VP1-aa 23-43 polypeptide is shown in SEQ ID NO.2. The polypeptide can carry FITC to enter a 293T cell, an HCT-116 cell, a 3T3 cell, an MDCK cell and an MSB1 cell within 30min; furthermore, the cell penetrating efficient is positively correlated to the concentration of the cell penetrating peptide; the key feature is that the polypeptide can efficiently enter suspension culture cells. The results of a laser scanning confocal microscope and a flow cytometry indicate that the cell penetrating efficiency of the CAV VP1-aa 23-43 polypeptide is over 2 times higher than that of a TAT oligopeptide at 10Mu M.

The invention discloses a cell culture layer which is prepared by the following methods: culturing BALB3T3 cells by adopting a DMEM culture medium, performing cell adherent culture until 70-80 percent of a culture bottle is full, and irradiating; performing X-ray irradiation by taking 3000cGy as a radiation dose; culturing by using an improved F culture medium after irradiation; culturing for 8 hours, and obtaining the culture, namely the cell culture layer. The cell culture layer can be used for culturing human primary cancer cells or cancer stem cells. The invention also discloses improvement of cell types of the cell culture layer as well as a preparation method of the cell culture layer. Compared with J2 cells, the 3T3 cells for preparing the culture layer are easily obtained, and the cell concentration and state can be easily controlled in the culture process. When the cell culture layer is used for culturing the human primary cancer cells or cancer stem cells, growth of the primary cancer cells can be well promoted, and human cancer stem cells in a higher ratio are obtained (the number of CD44 positive cells is improved by about 80 percent).

The present invention discloses one kind of human protein with 3T3 cell conversion promoting function, polynucleotides encoding the polypeptide and the recombinant process of producing the polypeptide. The present invention also discloses the agonist resisting the polypeptide and its treatment effect. The present invention also discloses the application of the polynucleotides encoding the human protein with 3T3 cell conversion promoting function.

The Rev peptide that binds to nucleophosmin/B23 with the highest affinity exhibits the greatest cytotoxicity on Ras-3T3 cells and inhibits tumor growth most effectively in nude mice. The efficiency of colony formation in soft agar of Ras-3T3 cells is significantly inhibited by treatment with Rev peptide. In addition, Rev peptide can potentiate the doxorubicin-induced decrease of cellular viability in U1 bladder cancer cells and inhibition of tumor growth in nude mice. Treatment of Rev peptide increases protein expression and transcriptional activity of p53 and inhibits the nucleophosmin/B23-mediated PCNA promoter activation. Peptides having high affinity of binding to molecular targets such as nucleophosmin/B23 represent a useful approach to anti-cancer biotherapeutics.

The invention relates to a use of a chicken anemia virus (CAV) VP1-aa 1-19 polypeptide as a highly efficient cell-penetrating peptide. The VP1-aa 1-19 polypeptide sequence is shown in SEQ ID NO. 2. The polypeptide can carry FITC and enter 293T cells, HCT-116 cells, 3T3 cells, MDCK cells and MSB1 cells in 30min, and the cell-penetrating efficiency is positively correlated with the concentration of the cell-penetrating peptide. The VP1-aa 1-19 polypeptide can efficiently enter suspension culture cells. The laser confocal microscope and flow cytometry result shows that the CAV VP1-aa 1-19 polypeptide has cell-penetrating efficiency significantly higher than that of a TAT short peptide.

The invention discloses a method for efficiently expressing an AFP3-PTEN fusion protein, and belongs to the technical field of biology. The method comprises the steps of cloning an HBx gene to a constructed expression vector, enabling the constructed vector to be transfected with human liver cancer cells, after expressing HBx proteins in the liver cancer cells, enabling the third structured domainof AFP and an overall-length PTEN gene to be connected to the expression vector and transfected into the human liver cancer cells expressing the HBx proteins, expressing the HBx proteins in the livercancer cells, promoting efficient secretory expression of a human AFP3-PTEN fusion protein through the HBx proteins, and concentrating and purifying culture mediums to obtain the AFP3-PTEN fusion protein. The AFP3-PTEN fusion protein expressed by the liver cancer cells can restrain cell proliferation and can enable the cell proliferation capacity of NIH 3T3 cells to be reduced by 15%-60%. But a reference protein AFP enables cell proliferation capacity to be obviously increased by 22-125%. Serum proteins do not have notable capacity for promoting NIH 3T3 cell proliferation. The obtained AFP3-PTEN fusion protein can be used for researching tumor treatment.

The invention discloses a cationic macromolecular proteolipid gene medicine carrier, a preparation method and an application, wherein the cationic macromolecular proteolipid gene medicine carrier is a macromolecular transferrin liposome prepared by transferrin-long-chain alkyl quaternary ammonium salt and a lipid ingredient in a mass ratio of (0.05 to 20): 1. The result of a gene transfection experiment indicates that the gene transfection efficiency of the macromolecular transferrin liposome in 293T cells, NIH-3T3 cells, liver cancer Huh-7 cells, SMMC-7721 cells, lung cancer A549 cells is equivalent to the gene transfection efficiency of a positive control, namely a cationic liposome Lipofectamine TM2000. The system of the cationic macromolecular proteolipid gene medicine carrier is good in biocompatibility, low in cytotoxicity, and capable of being used as an excellent non-viral gene medicine delivery carrier.

The present invention discloses a new kind of human protein with 3T3 cell transformation improving function, polynucleotides encoding this polypeptide and recombination process to produce the polypeptide. The present invention also discloses the agonist resisting the polypeptide and its treatment effect. The present invention also discloses the application of the polynucleotides encoding this human protein with 3T3 cell transformation improving function.

The invention discloses a method for establishing a transgenic removable human skin fibroblast feeder cell line. By utilizing a retrovirus-mediated gene transfer way, the production tool introduces areinforced green fluorescent protein gene, a telomerase reverse transcriptase gene and a herpes simplex virus thymidine kinase gene into human skin fibroblasts to establish a fluorescence-labeled immortalization-removable TERT + TK-D human feeder cell line. After being treated by mitomycin C, the established cell line serving as feeder cells is co-cultured with human limbal stem cells, and the culture result is compared with a culture result of 3T3 feeder cells. Transgenic fluorescence-labeled immortalization-removable human skin fibroblast feeder cells are expected to replace the 3T3 cells for corneal regeneration therapy.

A novel human protein with the function of promoting 3T3 cell transform, the polynucleotide for coding the peptide, the process for preparing said polypeptide by recombination, the antagon of said polypeptide and its medical action, and the application of said polynucleotide are disclosed.