14 results about "K562 cells" patented technology

The application provides a kind of HLA-A24:02 APC cell preparation method and application, it is related to cell preparation technical field.The present application constructs the APC cell (K562-HLA-A24:02) that can replace HLA-A24:02 subtype DC cell, fills the blank of HLA-A24:02 subtype special engineering APC cell, and the cell can realize the antigen presentation function consistent with natural HLA-A24:02 subtype DC cell, provides special tool cell for the immune research for the HLA subtype;The present application constructs APC cell with K562 cell line as base, K562 cell can be in vitro permanent passage amplification, without repeatedly separating and inducing DC cell from human peripheral blood, reduces the use amount of peripheral blood, reduces the raw material dependence and cost of cell acquisition, while avoiding the influence of individual difference of peripheral blood source on experimental results.

The present application belongs to the field of cell therapy and immunotherapy, and provides a genetically engineered human trophoblast, a preparation method and application thereof. The human trophoblast takes K562 cells as starting cells, and stably expresses membrane-bound interleukin 21, CD137 ligand and Delta-like ligand 1 after genetic engineering. The constructed K562 three-factor trophoblast can significantly improve the expansion efficiency, activation state and functional stability of NK cells and γδT cells. The synergistic mechanism includes enhancing the proliferation, cytotoxicity and stemness maintenance of NK cells and γδT cells through STAT3, NF-κB and Notch signaling pathways, respectively. The human trophoblast has the advantages of good expression stability, significant functional enhancement, and high activity after freezing and recovery.

The application discloses a kind of trichoderma fermentation processing indigo blue method, the product obtained and application.The method includes: preparation trichoderma seed liquid;After primary indigo blue sterilization, access seed liquid, fermentation 5-9 days under 25-30 DEG C, 120-180 rpm, avoid light, obtain secondary processing indigo blue.The application first utilizes trichoderma to carry out biological transformation to indigo blue, can significantly improve the content of its key anti-tumor component indigo carmine with specificity, and the increase is more than 150%.In vitro anti-tumor experiment shows that the inhibitory activity of secondary processing indigo blue prepared by the application to human leukemia K562 cells and human colon adenocarcinoma cell SW480 cell is significantly better than ordinary indigo blue.The product of the application can be used for preparing medicine for preventing and / or treating leukemia, colon adenocarcinoma, process controllable, suitable for industrialized production, and provides a new way for developing efficient indigo blue anticancer new drug.

The application discloses application of aza-3 in resisting erythroleukemia, wherein the aza-3 is a compound obtained by modifying a ring of aza-3 with 3alpha-O-(Boc-proline); the application proves that the aza-3 can obviously inhibit HEL and K562 cell proliferation, reduce splenomegaly, improve anemia indexes (RBC, HGB and HCT) and splenic and hepatic pathological infiltration, and has no obvious toxic side effects on main organs such as heart, lung and kidney through in-vitro MTT experiment and a Friend virus-induced erythroleukemia mouse model.

The present invention relates to the field of immune cell therapy, and specifically relates to a method for expanding NK cells and use thereof. The method comprises using OK-432 in combination with irradiated feeder cells, such as K562 cells, to expand and culture NK cells in vitro.

A method for producing γδ T cells modified by a chimeric antigen receptor, comprising: transfecting K562 cells with shFPPS targeted to FPP synthase by means of a lentiviral vector, such that the expression of FPPS in the K562 cells is lowered and a K562-shFPPS cell line with reduced FPPS expression is constructed; adding the K562-shFPPS cell line into a γδ T cell culture system for co-culturing with the γδ T cells, wherein it is found that the K562-shFPPS cell line can facilitate in vitro differentiation and proliferation of the γδ T cells; and adding a CAR-expressing lentiviral vector to the γδ T cell culture system comprising the cell line for co-culturing, wherein it is found that the K562-shFPPS cell line can further effectively improve the transfection rate of CAR genes. The provided solution effectively overcomes the technical challenge of the large-scale production of CAR-γδ T cells, and has a good application prospect.

The invention discloses a targeted protein degradation agent utilizing a ubiquitin-proteasome pathway as well as a preparation method and application of the targeted protein degradation agent, and belongs to the technical field of biological medicines. On the basis of a PROTAC (protein targeted degradation chimera) strategy and on the basis of imatinib, different Linkers are introduced to be connected with an E3 ubiquitin enzyme ligand Nutlin-3 derivative, a degradation tag is labeled on BCR-ABL cancer protein, and the degradation agent with the Bcr-Abl protein targeting capacity is constructed. A Western blot experiment shows that the targeted protein degradation agent shows a good degradation effect on the Bcr-Abl protein in K562 cells, and the compound WP shows a degradation effect on concentration dependence and time dependence of the Bcr-Abl protein. Tumor cell proliferation experiments show that the compound has certain inhibitory activity on K562 cells. Wherein when the Linker is dodecane-1, 12-diketone, the anti-proliferative activity is the best. An apoptosis experiment and a period experiment show that the targeted protein degradation agent can realize concentration-dependent promotion of cell apoptosis and retard cells in S and G1 / G0 periods.

The invention discloses an in-vitro amplification method for improving the purity of natural killer cells. The use combination of the cell factors is IL-2, IL-15, IL-18 and IL-21, the culture of the umbilical cord blood NK cells is optimized mainly from the aspects of the factor combination used for culturing the NK cells, the use concentration and the like, CD3-CD56 + positive indexes all reach 80% or above, and the amplification rate reaches 100 times or above. The method is also suitable for large-scale culture of peripheral blood NK cells, the CD3-CD56 < + > positive rate reaches 99.0% or above, the amplification rate reaches 1000 times or above, and the killing rate of K562 cells is 30% or above when the effect-target ratio is 1: 1.

The present disclosure provides a fully human therapeutic monoclonal antibody or fragment thereof, which is a fully human antibody specifically binding to the RBD region of SARS-CoV-2 S protein obtained from isolating a single B cell clone from a convalescent blood sample of a Covid-19 patient, which does not produce ADE effect on SARS-CoV-2 infection of THP-1 cells and K562 cells. The monoclonal antibody has high affinity to the RBD of SARS-CoV-2 S protein, with a KD value of 5.0 x 10 ‑9 M; and the monoclonal antibody has blocking activity to the binding of the RBD of SARS-CoV-2 S protein to ACEII, with an IC50 value less than 50 nM.

This invention discloses a method for detecting the killing effect of multiple HLA-specific T cells, belonging to the field of biotechnology. This invention enables T2 cells to express various HLA-I molecular subtypes. After the receptor is expressed, antigen fragments loaded with various HLA-I molecules can be captured in the culture supernatant for in vitro evaluation of the killing effect of HLA-I-adapted antigen-specific T cells, thus expanding the application of T2 cells in in vitro killing effect evaluation. Simultaneously, it avoids the enormous workload and high cost of using the K562 cell line to construct a cell line for each HLA subtype-specific antigen fragment during the HLA subtype construction process.

The invention relates to the technical field of medicines, in particular to application of actinomycin V in preparation of a medicine for treating chronic myelogenous leukemia. In the aspect of cell function, the Act-V shows comprehensive anti-CML activity, not only can inhibit K562 cell proliferation in a time and dose dependent manner, but also can effectively inhibit colony forming ability, reduce the number and diameter of formed tumor spheres and lower the sphere forming frequency. Experiments prove that the Act-V further weakens the dry characteristics of tumors by down-regulating the expression of dry markers such as CD133, CD44, ALDH1A1 and the like. Besides, the Act-V can significantly inhibit the invasion and migration ability of CML cells, and western blot experiment results prove that the Act-V down-regulates the expression of interstitial markers N-Cadherin and Vimentin, up-regulates the expression of an epithelial marker E-Cadherin and inhibits the invasion and migration ability, so that the Act-V has wide application scenarios.

The application provides a method for constructing 1,2-cis fucose glycoside bond with high stereoselectivity and application in preparation of 13-sugar repeating units and 9-sugar substructures of angelica polysaccharide APS-1II, and evaluation of anti-leukemia activity of the 13-sugar and 9-sugar, and belongs to the technical field of polysaccharide preparation; the application realizes a promoter composition of 1,2-cis fucose glycosylation reaction with high stereoselectivity, which comprises iodotrimethylsilane (TMSI) and triphenyl phosphine oxide (Ph3PO), and the donor is fucosyl N-phenyl trifluoroacetyl imidate (PTFAI) with acyl substitution at O-3 and / or O-4 positions. The 13-sugar repeating units and 9-sugar substructures of angelica polysaccharide APS-1II provided by the application have significant inhibitory effect on human leukemia K562 cells and mouse leukemia L1210 cells.

The invention discloses a construction method and application of a membrane-bound IL-21 K562 cell strain, a human IL-21 mature sequence is fused with an IgG4 hinge region, a human immune globulin gamma-4 chain CH2 / CH3 region and a CD4 transmembrane structural domain through genetic engineering to construct an mbIL-21 fusion gene, and the mbIL-21 fusion gene is cloned to a PT-mnud carrier by using BamH I / Xho I double enzyme cutting sites to obtain a recombinant plasmid PCDH-mbIL-21. HEK 293T cells are transfected through a lentivirus packaging system, high-titer lentivirus particles are prepared, and the cell strain stably expressing mbIL-21 is obtained through puromycin screening after K562 cells are infected. The mbIL-21 K562 cell strain constructed by the invention has the characteristics of long acting, targeting and stable expression, and provides a powerful tool for basic research and clinical transformation of immunotherapy.

The present invention provides an application of thyroid hormones and thyroid hormone analogues to preparation of drugs for treating sickle-cell disease, particularly an application to preparation of drugs for improving the expression quantity of ζ-globin. During the differentiation of K562 cells, thyroid hormone analogue (Triac) can significantly up-regulate the expression of ζglobin gene (HBZ) by 50 folds or above. The expression of ζ-globin gene (hbae5) can also be up-regulated by 30-70 folds in zebrafish treated with thyroid hormones and thyroid hormone analogues. Therefore, according to the present invention, the expression of ζ-globin gene can be significantly activated by thyroid hormones and thyroid hormone analogues which may develop new potential therapies for patients with sickle-cell disease. It provides an economical, safe and effective method for treating sickle-cell disease and can be widely used.