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192 results about "K562 cells" patented technology

K562 cells were the first human immortalised myelogenous leukemia cell line to be established. K562 cells are of the erythroleukemia type, and the cell line is derived from a 53-year-old female chronic myelogenous leukemia patient in blast crisis. The cells are non-adherent and rounded, are positive for the bcr:abl fusion gene, and bear some proteomic resemblance to both undifferentiated granulocytes and erythrocytes.

Method for amplifying NK cells of human beings under condition of in vitro culture

The invention discloses a method for amplifying NK cells of human beings under the condition of in vitro culture, which is characterized in that a peripheral blood mononuclear cell (PBMC) is taken asthe original culturing material, and the PBMC is cultured together with an engineering cell which is built by adopting the genetic engineering method and is used for stimulating the growth of the NK cell. The built engineering cell which is used for stimulating the growth of the NK cell expresses several cytokines (IL-2, IL-12, IL-15, IL-18, 4-1BB) which can promote the growth of the NK cell on the surface of the K562 cell; after irradiation and inactivation with gamma-rays, the engineering cell is cultured with the PBMC in vitro, as a result, the amplification effect of the stimulation methodin the invention is hundreds of times stronger than that of the currently universal method in which soluble cytokines are added purely to the culture solution; and after cultured for 3 weeks, the non-NK cells in the PBMC are mainly dead and disappeared, the NK cells are proliferated in great quantity, the purity of the NK cells reaches over 96% and the total number of the NK cells is amplified byover 1500 times.
Owner:JIANGMEN LUOSEN BIO PHARMA

Method for in-vitro amplification of NK cells

The invention relates to a method for in-vitro amplification of NK cells, and in particular relates to a method for massive in-vitro amplification of NK cells, wherein the method comprises the following steps of: a, inoculating a peripheral blood mononuclear cell in a CD3McAb and CD226McAb pre-coated culture bottle for coculture; b, adding 1L-2 and 1L-18, coculturing for 72hours to stimulate amplification of NK cells; c, transferring the NK cells, K562 cells after lethal treatment and a serum-free medium containing 1L-2 and 1L-18 in a cell culture bag for coculture; and d, collecting the NK cells. According to the method for in-vitro amplification of the NK cells, two antibodies CD3McAb and CD226McAb are simultaneously coated, so the cell factor synthesis and ADCC effect are promoted, and killing toxicity of the NK cells is remarkably improved; the activation and amplification on the NK cells are achieved just by the 1L-2 and 1L-18 cell factors, so the amplification multiple and cell toxicity of the NK cells are guaranteed, and the cost of cell culture is reduced.
Owner:SHANGHAI CLAISON BIOTECH

Method for amplification and activation of NK cells by K562 cells

ActiveCN103232973AHelp identify and killHelp activate recognition and killMammal material medical ingredientsBlood/immune system cellsNatural Killer Cell Inhibitory ReceptorsCD86
The invention discloses a method for amplification and activation of NK cells by K562 cells. The method comprises that through synergism of K562 cells transfected by transmembrane interleukin 21, CD14, CD19, CD86 and CD137, and low-concentration interleukin 2, NK cells are subjected to directed amplification and activation. Compared with the existing similar compounds, the compound provided by the invention has a stronger lymphocyte amplification and activation capability and higher efficiency. The method has wide prospects in immunological therapy.
Owner:杭州中赢生物医疗科技有限公司

Method for preparing NK (natural killer) cell

The invention provides a method for efficiently preparing an NK cell, which can improve the multiplication rate and the purity of the NK cell through combination of stimulation effects of cell factors and feeder cells. The method is mainly characterized by comprising the steps as follows: NCR3LG1 and m IL-15 are transfected to a K562 cell simultaneously, m IL-15 can be used for adjusting activation and multiplication of the NK cell, NCR3LG1 serving as a ligand of NKp30 which is one of main activated receptors on the surface of the NK cell can effectively stimulate activation of the NK cell, and NCR3LG1 and m IL-15 have a synergistic effect; and PBMC(peripheral blood mononuclear cells) can be multiplied over 500 times in 21 cultivation days through stimulation of factors of freely added IL-2, IL-21 and the like, and a proportion of CD3-CD56+NK cell exceeds 70%; and up to now, a research report that NCR3LG1 and m IL-15 are transfected to the feeder cells simultaneously and jointly stimulate and activate the NK cell in combination of free cell factors is absent. The invention firstly provides a method for jointly cultivating and preparing the NK cell in combination of the feeder cells and the free factors.
Owner:青岛麦迪赛斯生物科技有限公司

Artificial antigen presenting cell and application thereof in NK (natural killer) cell amplification

The invention provides an in vitro amplification method for efficient and highly cytotoxic natural killer (NK) cells. Novel artificial antigen presenting cells 4-1BBL-mIL-21-aAPC, such as 4-1BBL-mIL-21-K562 cells and the like, are constructed through stably expressing 4-1BB ligands (4-1BBL) and membrane immobilized interleukin 21 (mIL-21) on the surfaces of cell membranes, and by using the novel artificial antigen presenting cells as feeder cells for amplification, the NK cells are directly amplified from peripheral blood lymphocytes. Flow cytometry, cytotoxicity test and the like suggest that the amplified cells NK have high purity and strong cytotoxicity and have obvious killing effect on tumor cells.
Owner:SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE

Gene engineering cell and application thereof in NK (Nature Killer) cell proliferation

The invention discloses a gene engineering cell and application thereof in NK (Nature Killer) cell proliferation, and provides an in vitro proliferation method of an NK cell with high efficiency and high cytotoxicity. A 4-1BB ligand (4-1BBL) and a membrane fixed interleukin 21 (mbIl-21) are stably expressed on the surface of a cell membrane by adopting a genetic engineering technology for constructing 4-1BBL-mbIl-21-gene engineering cells (4-1BBL-mbIl-21-Gene Engineering Cells, 4-1BBL-mbIl-21-GEC), such as 4-1BBL-mbIl-21-K562 cells, to be used as feeder cells for proliferation, and the NK cells are directly proliferated from peripheral blood monouclear cells, thus the operation is simpler and easier. Studies such as cell counting, flow cytometry and cytotoxicity tests indicate that high cell proliferation times, high NK cell purity, strong cytotoxicity and remarkable cancer cell killer effect are achieved.
Owner:SHANGHAI JIAOTONG UNIV SCHOOL OF MEDICINE

Method for high-efficient expansion and cryopreservation of NK (Natural Killer) cells and application of method

The invention belongs to the field of cell engineering, in particular to a method for high-efficient expansion and cryopreservation of NK (Natural Killer) cells and an application of the method. According to the invention, fusion genes are subjected to gene recombination to an AAVS1 (Adeno-Associated Virus Integration Site 1) transposon through a plasmid, and the freezing / thawing step is introduced to the NK cell expansion step taking a modified K562 cell line as a basis in the intermediate stage of cell expansion; finally, the NK cells are expanded and saved. A K562-mbIL15-41BBL-mbIL21 cell line developed by the integration technology of site specific genes is used, and the activity, purity and cytotoxicity of the NK cells are high to realize high expansion efficiency of the NK cells to improve NK expansion. The method for the high-efficient expansion and cryopreservation of the NK (Natural Killer) cells and the application of the method for the high-efficient expansion and cryopreservation of the NK (Natural Killer) cells disclosed by the invention have great market prospects and economic values.
Owner:杭州朔溪生物医药有限公司

Method for amplifying and activating lymphocyte by using CD8 alpha-interleukin 21-CD137 compound

The invention relates to the field of immunology, in particular to a method for amplifying and activating a natural killer (NK) cell into a lymphokine-activated killer (LAK) cell by using a CD8 alpha-interleukin 21-CD137 compound. The method disclosed by the invention comprises the following steps of: forming the CD8 alpha-interleukin 21-CD137 compound by using CD8 alpha, interleukin 21 and a CD137 functional polypeptide, making an exogenous expression vector enter a host K562 cell, then activating a promoter and culturing a cell to obtain a cell for expressing a transmembrane interleukin 21-CD137 compound; and purifying the compound in the conventional way, and amplifying and activating a lymphocyte by using the purified compound to generate the LAK cell. The method has the advantage that: the LAK cell cultured and amplified by using the transmembrane interleukin 21-CD137 compound and a small dose of interleukin 2 is used for enhancing the immunity of a patient to help the patient resist tumors, viruses and bacteria. The method has a wide clinical using prospect.
Owner:杭州中赢生物医疗科技有限公司

Long-chain non-coding RNA IncRNA-BcrAR and application thereof in cell canceration resistance

The invention relates to a long-chain non-coding RNA IncRNA-BcrAR and application thereof in cell canceration resistance. IncRNA-BcrAR is in low-level expression in a human chronic myelogenous leukemia cell line K562 with positive Bcr-Abl, the K562 cell line with overexpressed IncRNA-BcrAR is constructed, the action of IncRNA-BcrAR on Bcr-Abl induced cell neoplastic transformation is observed, experiments prove that the IncRNA-BcrAR overexpression can obviously promote K562 cell apoptosis induced by Imatinib (therapeutic drug of Abl positive leukemia patients) and can remarkably inhibit tumor growth induced by K562 cells in a naked mouse body; and besides, the IncRNA-BcrAR overexpression can remarkably promote A-MuLV transformed mouse leukemia cell BC44 apoptosis induced by Imatinib. The IncRNA-BcrAR has important effect on Bcr-Abl and v-Abl mediated cell canceration resistance, and the long-chain non-coding RNA IncRNA-BcrAR provides new molecular marker and drug target for diagnosis and treatment of Abl induced leukemia.
Owner:FUJIAN AGRI & FORESTRY UNIV

Antisense oligonucleotide for tiny RNA-21 seed sequence and application thereof

The invention discloses an antisense oligonucleotide for tiny RNA-21 seed sequence and an application thereof and belongs to the field of medicine preparation. According to the research, tiny antisense nucleic acid t-antimiR-21 of 8 basic groups is designed and synthetized for miR-21 seed sequence. It shows through a t-antimiR-21 transfection cell experiment that t-antimiR-21 targets miR-21 of K562 cell and RPMI-8266 cell strain, cell growth is inhibited and cell apoptosis is obviously increased. It shows that lymphoma cell growth can be effectively inhibited through interfering with miRNA-21 expression by t-antimiR-21. Meanwhile, it shows that miRNA-21 might be used as a potential target for lymphoma gene therapy. t-antimiR-21 can be used in the preparation of an antineoplastic medicine, especially in the preparation of a medicine against chronic granulocytic leukemia and multiple myeloma.
Owner:广州安镝声生物医药科技有限公司

New medical use of cucurbitacin

The invention discloses a new medicament application of cucurbitacins, namely an application of mixing effective monomers of cucurbitacins in medicament for treating leukaemia or an application of effective monomers of cucurbitacins to independently treating leukaemia, belonging to the technical field of medicament. The cucurbitacins with nano-mole concentration can obviously inhibit multiplication of leukaemia HL60 and K562 cells; the cucurbitacins have no cell toxic effect to human leukaemia cells in a multiplication inhibiting concentration range; and the multiplication inhibiting effect to leukaemia cells in vitro is stronger than that to solid tumour cells. Further researches suggest that the cucurbitacins with micro-mole concentration can induce apoptosis of leukaemia HL60 cells by increasing cucurbitacin concentration. Anti-tumour researches in vivo suggest that the cucurbitacins can prolong survival time of a small mouse P388 leukaemia animal model. The effective monomers of cucurbitacins in resisting leukaemia have obvious effect, low toxicity, long lasting drug administration, can be used for both chemotherapy and radiotherapy, and are promising to be further developed into the medicament for treating leukaemia.
Owner:SHENYANG PHARMA UNIVERSITY

Preparation method for efficiently amplifying NK cells by utilizing trophoblasts

The invention relates to the field of gene engineering and cytobiology, in particular to a preparation method for efficiently amplifying NK cells by utilizing trophoblasts. The construction method comprises the following steps: (1) constructing a pHR-mbIL-21 plasmid vector, a pHR-4-1BBL plasmid vector and a pHR-MICA plasmid vector; (2) preparing recombinant lentiviruses by using pHR-mbIL-21, pHR-4-1BBL and pHR-MICA plasmid vectors respectively; (3) preparing K562-mbIL-21-4-1BBL-MICA trophoblasts; (4) extracting PBMC cells; and (5) carrying out in-vitro amplification on NK cells. The inventionprovides the preparation method of NK cells. The method employs the step of independently constructing plasmid vectors for expressing mbIL-21, 4-1BBL and MICA molecules, K562 is infected with recombinant lentivirus, the NK cells can be prepared, the preparation method overcomes the defects in the prior art, the K562 cells simultaneously expressed by IL-21, 4-1BBL and MICA molecules are used as trophoblasts for amplification culture of the NK cells, the amplification multiple of the NK cells reaches up to 890 times, the purity of the prepared NK cells reaches 92.2%, and the repeatability of theamplification multiple between different PBMC cells is good.
Owner:山东德升生物工程有限公司

Artificial antigen submit cell and preparation method thereof

The invention discloses an artificial antigen submitting cell and preparing method, which comprises the following steps: choosing human chronic granular leukocyte leukemia bacterial strain K562 cell as carrier; transfer-dying and expressing eucaryon expressing carrier of CD32a; expressing CD32 molecular on the surface of K562 cell stably through G418 screening and flowed cell sorting device sorting; forming K32 cell; producing eucaryon expressing carrier of double expressing CD86 and 4-1BBL co-simulating signal; proceeding homomycin screen and sort with flowed cell device for the K32 cell; getting stable expression K32 cell of CD86 and 4-1BBL molecule; getting the end product. This invention possesses low cost and good repeatability, which can inhibit die effectively.
Owner:YANGZHOU UNIV

Preparation method and application of N-substituted-3,5-dibenzal piperidine-4-one

The invention belongs to a lead compound of a novel drug for preventing leukaemia K562 cell proliferation and relates to a preparation method and application of N-substituted-3,5-dibenzal piperidine-4-one which can effectively inhibit leukaemia K562 cell proliferation. The preparation method comprises the following steps of: performing Michael addition reaction for substituted amine and methyl acrylate to prepare a yellow oily object N,N-di(beta-methyl propionate) substituted amine; performing Dieckmann condensation under effect of sodium alkoxide and performing hydrolysis and decarboxylationunder effect of acid to obtain a yellow oily object N-substituted piperidine-4-one; and dehydrating the product obtained to obtain the N-substituted-3,5-dibenzal piperidine-4-one with the general formula (I). The product of the invention has higher inhibition activity to eukaemia K562 cell proliferation and the method has the advantages of simple process and easy production.
Owner:SHANGHAI NORMAL UNIVERSITY

Method for producing chimeric antigen receptor modified gamma delta T cell

The invention discloses a method for producing a chimeric antigen receptor modified gamma delta T cell. The method ensures that shFPPS targeted to FPP synthetase transfers a K562 cell through a lentiviral vector, the expression quantity of FPPS in the K562 cell is lowered, and a K562-shFPPS cell line with lowed FPPS expression quantity is constructed. According to the method, a K562-shFPPS cell line is added into a gamma delta T cell culture system to be co-cultured with gamma delta T cell, and the fact that the K562-shFPPS cell line can promote the in vitro differentiation and expansion of the gamma delta T cell is found. According to the method, the lentiviral vector expressing CAR is further added into the gamma delta T cell culture system containing the cell line, so that co-culturingis performed, and the fact that the K562-shFPPS cell line can further effectively improve the transfection efficiency of CAR gene is found. The method effectively solves the technical bottleneck of mass production of CAR-gamma delta T cell, and has an excellent application prospect.
Owner:HEBEI SENLANG BIOTECH CO LTD

Application of Trichoderma pseudokoningii extracellular polysaccharides having anticancer activities

The invention relates to an application of Trichoderma pseudokoningii extracellular polysaccharides having anticancer activities. The Trichoderma pseudokoningii extracellular polysaccharides having the anticancer activities can reduce the vitality of K562 cells in vitro when they are applied to tumor growth inhibition (auxiliary) medicines, so intracellular active oxygen and free calcium ion concentrations are improved, the expression abundance of mRNA of p53 and Bax genes is improved, the expression of the Bc1-2 transcription level is reduced, the ectropion of cell membrane phosphatidylserine of the human chronic granulocyte leukemia cells K562 is induced, and the nuclear polycondensation and the fragmentation of K562 are induced to generate apoptotic bodies and apoptosis; and in-vivo administration can increase weights of tumor bearing mice and alleviate growth speeds and weights of tumors of the tumor bearing mice, thereby tumor cell killing and tumor growth inhibiting purposes are reached.
Owner:博德生物技术(德州)有限公司

Preparation of immunoliposome of CD19 monoclonal antibody and application

The invention provides an immunoliposome of a CD19 monoclonal antibody. First, a film dispersion method is adopted to prepare the cubical space stable norcantharidin immunoliposome, wherein the mole ratio of soy bean lecithin, cholesterol, methoxyl-polyethylene glycol 2000-phosphatidyl ethanolamine is 2:1:0.01, the mass ratio of norcantharidin immunoliposome and soy bean lecithin is 1:20, and entrapment efficiency of high-performance liquid detection NCTD is 46.5+ / -2.21%. The research proves that 2E8-SL-PE can be combined with Nalm-6 and Raji cell specifically, the positive rates thereof are respectively 89.44% and 88.73%, the 2E8-SL-PE is almost not combined with Molt-3 and K562 cell with the positive rates being only 1.95% and 1.39%, which shows that the immunoliposome has obvious target recognition effect. The norcantharidin immunoliposome can be applied in preparing specific target killer B lymphocyte leukemia and stem cell drugs thereof.
Owner:杭州永申生物科技有限公司

Preparation method and application of autologous natural killer cell proliferation

The invention provides a preparation method of autologous natural killer cell proliferation and culture. The preparation method of the autologous natural killer cell proliferation and culture is characterized in that mononuclear cells of a cancer patient are amplified and activated to obtain natural killer cells after the mononuclear cells are transfected by 3- phosphoinositide-dependent protein kinase-1 and CD122 and under the effect of K562 cells which are expressed stably serve as trophoblast cells and interleukin 2, proliferation times of the natural killer cells are above 2500, phenotype of CD16+ / CD56+ is above 90%, and in-vivo and in-vitro tests show that the natural killer cells have an anti-tumor and have quite high killer toxicity. The invention also provides a kit which is used for autologous natural killer cell proliferation and culture and is obtained by using the preparation method. The kit has an efficient anti-tumor function.
Owner:ZICHENG RUISHENGHUI BEIJING BIOTECH DEV CO LTD

Tyrosine kinase inhibitor and preparation method and use thereof

The invention relates to a tyrosine kinase inhibitor and a preparation method and use thereof, and belongs to the technical field of pharmaceutical chemistry. The tyrosine kinase inhibitor having the structural features shown in the general formula I, or its pharmaceutically acceptable salts or stereoisomers can effectively inhibit tyrosine kinase activity, and can inhibit kinases such as DDR1, DDR2, Abl, Src, Btk and Kit. Compared with a positive contrast dasatinib, the tyrosine kinase inhibitor has higher half inhibitory concentration or the same half inhibitory concentration, and especially, aiming at DDR1, DDR2, Src, Btk and Kit, the compound 8j has lower K562 cell half inhibitory concentration. The tyrosine kinase inhibitor has good enzyme inhibitory activity and cell activity and has a large application prospect.
Owner:GUANGZHOU INST OF BIOMEDICINE & HEALTH CHINESE ACAD OF SCI

Tetrahydropyridopyridone derivative as well as preparation method and application thereof

The invention provides a tetrahydropyridopyridone derivative as shown in a general formula (I), and a preparation method thereof. The method comprises the following steps of: preparing N,N-bi(methoxycarbonyl group) substituted benzylamine (b) by carrying out Michael addition on substituted benzylamine (a) and methyl acrylate, carrying out Dieckmann condensation on (b) under the action of sodium alcoholate, subsequently carrying out hydrolysis and decarboxylation under the action of acid so as to obtain N-substituted benzyl-4-piperidone (d), carrying out an aldol condensation reaction on (d) and bimolecular aromatic aldehyde so as to obtain N-substituted-3,5-bi(substituted benzylidene piperidine-4-ketone (e), and refluxing and heating (e) malononitrile and ammonium acetate in ethanol so as to obtain a final product as shown in the general formula (I). The tetrahydropyridopyridone derivative is simple in process and convenient to produce in scale, and the compound (I) has a good inhibition effect on multiplication of leukemia K562 cells, ovarian cancer HO-8910 cells and liver cancer SMMC-7721 cells. Therefore, the invention further provides an application of the compound as shown in the general formula (I) in preparing medicaments for preventing multiplication of the leukemia K562 cells, the ovarian cancer HO-8910 cells and the liver cancer SMMC-7721 cells.
Owner:SHANGHAI NORMAL UNIVERSITY +1

Method for identifying differential marker of IC50 dose of vitamin C for RAW264.7 and K562 cells

The invention discloses a method for identifying a differential marker of IC50 dose of vitamin C for RAW264.7 and K562 cells by applying 1H NMR technology and metabonomic methodology. The method comprises the following steps: (1) determining the IC50 dosage of vitamin C for RAW264.7 or K562 cells; (2) preparing a cell extract; (3) performing nuclear magnetic resonance pretreatment; (4) performing 1H nuclear magnetic resonance detection; (5) screening differential metabolites; and (6) performing metabolism pathway analysis. The method studies influence of different cell differential metabolites and metabolic pathway by applying H-nuclear magnetic resonance technology in combination with metabonomic methodology, and the basic research provides theoretical basis for studying the method for analyzing sub-toxicity of vitamin C for RAW264.7 or K562 cells.
Owner:SHANDONG NORMAL UNIV

Method for preparing CD3+CD56+cells through high-killing K562cells from perinatal placental blood

ActiveCN103173409AGuaranteed stabilityThe collection process is convenient and safeBlood/immune system cellsBiologyK562 cells
The invention provides a method for preparing CD3+CD56+cells through high-killing K562 cells from perinatal placental blood. The method comprises the step of: separating mononuclear cells from the perinatal placental blood so as to carry out induction differentiation to form CD3+CD56+cells. By utilizing the method, the preparation period of the cells can be shortened, and moreover, the multiplication capability of the cells is improved.
Owner:JIANGSU HEZE STEM CELL GENE ENG CO LTD

Phenylbutyryl curcumin derivate and application thereof in anti-tumor drug preparation

The invention relates to phenylbutyryl curcumin derivate and application thereof in anti-tumor drug preparation, in particular to 4-(di(2-chloroethyl) amino) phenylbutyryl curcumin, 4,4'-(di(2-chloroethyl)amino) diphenylbutyryl curcumin, pharmaceutically acceptable salts and a preparation method thereof, and applications of 4-(di(2-chloroethyl) amino) phenylbutyryl curcumin, and 4,4'-(di(2-chloroethyl)amino) di phenylbutyryl curcumin in anti-tumor drug preparation. The phenylbutyryl curcumin derivate can be used for (but not limited to) preparing drugs for treating leukemia, skin cancer, gastric cancer, colon cancer, liver cancer, breast cancer or prostatic cancer. The derivate has obvious inhibition on a plurality of animal tumor cell transplanting modules, especially for the human chronic granulocytic leukemia model constructed by NOD-SCID mice inoculated with the human chronic granulocytic leukemia K562 cell stain, the life prolonging rate is obviously enhanced, and the serious toxicity to mice does not exist.
Owner:FUJIAN MEDICAL UNIV

Hydroxyproline peptoid derivative and preparation method and application thereof

InactiveCN109796439AHas inhibitory activityStrong inhibitory activityOrganic chemistryAntineoplastic agentsBcr-Abl tyrosine-kinase inhibitorBcr abl kinase
Disclosed are a hydroxyproline peptoid derivative and a preparation method and application thereof. With biphenyl pyridine as a hinge region binding fragment, by adopting a design strategy of a fragment drug and introducing L-hydroxyproline as a flexible Linker, a peptoid micromolecule-like compound library with kinase inhibition activity is constructed, and through screening of ADP-Glo and otheractivity tests, a peptoid tyrosine-like kinase inhibitor which has Bcr-Abl kinase inhibition activity and prevents the proliferation of tumor cells is found. The compound can be used for preparing anti-tumor drugs and has the activity of inhibiting Bcr-Abl and Bcr-AblT315I kinases and activity of inhibiting the cell proliferation of K562 cells. By introducing the L-hydroxyproline, the structural diversity of the Bcr-Abl inhibitor is expanded, the activity result shows that the introduction of proline has a certain effect on the inhibition activity of the compound, and the proline can serve asa novel pharmacological-effect fragment of the Bcr-Abl tyrosine kinase inhibitor.
Owner:XIAN CENT HOSPITAL

Composition for autologous lymphocyte culture, culture solution and application of composition

The invention discloses a composition for autologous lymphocyte culture, a culture solution and application of the composition. The active ingredients of the composition for autologous lymphocyte culture are composed of plasma, an anti-human CD3 monoclonal antibody, IL-2, IL-7, IL-15 and PHA-P. Compared with a composition for lymphocyte culture as a control, the content of a T lymphocyte subset obtained by using a culture medium containing the composition for induced culture of peripheral blood mononuclear cells is significantly increased; the number of lymphocytes is also significantly increased after 14 days of culture, and the number is increased by 1.15 times; and the killing activity of the lymphocytes on K562 cells is also significantly enhanced, and the killing activity is enhancedby 1.6 times. The composition for autologous lymphocyte culture can be used for specific induced amplification of mononuclear cells to produce autologous lymphocytes with high purity and strong biological activity.
Owner:北京奥康华医学检验所有限公司

Method for efficiently amplifying NK cells in vitro

The invention provides a method for amplifying NK cells by utilizing gene modified K562 trophoblast cells. IL-21, IL-27 and CD70 genes are transfected into K562 cells through a lentivirus reprint system, so that K562 engineering cells capable of stably expressing IL-21, IL-27 and CD70 protein can be obtained; the K562 engineering cells are mixed with PBMC to perform cultivation after being inactivated, so that the NK cells can be amplified in a serum-free medium under the presence of IL-2, and the amplification multiples can be about 800-1,100; the effective purity of the NK cells can reach 91%, and the secretion volume of IFN-gamma can be five times that of the NK cells cultivated by common methods; and the killing rate of the NK cells on hepatoma carcinoma cell HepG2 can reach 88.5% under the condition that effector-target ratio is 10: 1, which is about 20% higher than that of the NK cells cultivated under normal conditions, and therefore, the NK cells cultivated by the method have stronger killing abilities.
Owner:山东省齐鲁细胞治疗工程技术有限公司 +1

Genetically engineered cell and method for efficiently amplifying NK cells in vitro

The invention relates to specifically relates to a genetically engineered cell and a method for efficiently amplifying NK cells in vitro by using the genetically engineered cell, belonging to the field of immunology. According to the invention, a Piggybac transposon system is utilized to construct a K562 engineered cell that realizes transmembrane stable expression of IL-15, IL-18 and 4-1BBL; andthe method for amplifying and activating NK cells based on the genetically engineered K562 cell is provided in virtue of an in-vitro cell stimulation technology. The genetically engineered cell can realize simultaneous transmembrane high-expression of IL-15, IL-18 and 4-1BBL. The NK cell amplification method of the invention is simple to operate and low in cost; the amplified NK cells have the advantages of high purity, large quantity, good killing activity, etc.; and the NK cell amplification method is suitable for large-scale production of NK cells and lays a good foundation for the clinicalapplication of adoptive immunotherapy of NK cells.
Owner:武汉赛云博生物科技有限公司

Method for detecting killing activity of human (gamma)(delta)T cells against K562 cell strain

The invention belongs to the technical field of cellular immunology, and particularly relates to a method for detecting the killing activity of human (gamma)(delta)T cells against a K562 cell strain, which is used for detecting the prepared human (gamma)(delta)T cells. The method comprises the following steps: preparing the human (gamma)(delta)T cells; fetching the K562 cell strains of the logarithmic phase as target cells, and adjusting the cell density to 1*10<5> / ml, 5*10<4> / ml and 2.5*10<4> / ml; paving the target cells in a 96-hole culture plate with 100mu l per hole, and culturing for 24 hours; fetching the prepared human (gamma)(delta)T cells, and adjusting the cell density to 1*10<6>ml; adding the human (gamma)(delta)T cells into the 96-hole culture plate, wherein the effect-target ratios are 10:1, 20:1 and 40:1 respectively; and setting 3 parallel holes for each density, setting blank controls respectively and calculating the killing activity. By adopting the method provided by the invention, the toxicity of the human (gamma)(delta)T cells is guaranteed.
Owner:SHENZHEN HORNETCORN BIOTECH
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