183 results about "CD16" patented technology

The invention provides a natural killer cell, NK-92, modified to express an Fc receptor on the surface of the cell, such as CD16 (FcγRIII-A), or other Fcγ or Fc receptors. The modified NK-92 cell can be further modified to concurrently express an associated accessory signaling protein, such as FcεRI-γ, TCR-ζ, or to concurrently express interleukin-2 (IL-2) or other cytokines. Additional methods are disclosed for various assays, assessments, and therapeutic treatments with the modified NK-92 cells.

Methods for the identification and isolation of multipotent cells from non-osteochondral mesenchymal tissue. Specifically, this invention relates to an adult multipotent cell or a cell population or composition comprising said cell, isolated from non-osteochondral mesenchymal tissue, characterized in that the cell is positive for the following markers: CD9, CD10, CD13, CD29, CD44, CD49A, CD51, CD54, CD55, CD58, CD59, CD90 and CD105 and because it lacks expression of the following markers: CD11b, CD14, CD15, CD16, CD31, CD34, CD45, CD49f, CD102, CD104, CD106 and CD133.

A method is provided for enhancing ADCC in an individual in need thereof, comprising the administration of T lymphocytes expressing a CD16-like receptor in said individual. Said method for enhancing ADCC may be used to treating cancers, autoimmune diseases or infections.

The invention discloses a method for simultaneous and efficient amplification of CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells. The method comprises the steps as follows: the concentration of separated PBMC (peripheral blood mononuclear cells) is adjusted by a serum-free medium containing autologous plasma, an Anti-CD16 antibody, IL-2 and IL-15 are added, and then the mixture is transferred into a T175 culture flask for culture; an Anti-CD3 antibody and an Anti-CD137 antibody are added; a serum-free medium containing the autologous plasma, IL-2 and IL-15 is supplemented every two days according to the cell growth condition; the cell concentration is controlled to be about 1.5*10<6>/ml; and after culture is performed for 14-21 days, large quantities of high-purity CD<3+>CD<56+>CIK cells and CD<3->CD<56+>NK cells can be obtained simultaneously, and the total cell quantity can reach an effective value of the cell quantity required for adoptive cellular immunotherapy clinically for tumor. The method for simultaneous and efficient amplification of the CD<3+>CD<56+>CIK cells and the CD<3->CD<56+>NK cells is simple, convenient, effective and high in cell killing activity.

The invention relates to the field of stem cell biology, specifically to a method for differentiating human pluripotent stem cells into natural killer cells and an application. The invention disclosesa pluripotent stem cell-derived natural killer cell. The pluripotent stem cell-derived natural killer cell expresses CD56, Nkp30, Nkp44 and Nkp46, and also expresses markers CD16 and CD94 of a maturenatural killer cell. The invention also discloses a method for preparing the natural killer cell. The method comprises the following steps: S1, formation of an embryoid body; S2, differentiation of the embryoid body into hematopoietic progenitor cells; S3, differentiation of the hematopoietic progenitor cells into NK cells; and S4, maturation and expansion of the NK cells. With the differentiation method provided by the invention, the pluripotent stem cells can be rapidly, efficiently, simply and conveniently induced to be differentiated into the natural killer cells with low cost on the basis of a culture medium with definite components and an optimized cell factor combination.

A human monocyte cell characterised by the expression of the following markers: Tie2, CD16 and CD14.

The invention relates to a method for abundantly amplifying NK (Natural Killer) cells from a mononuclear cell of peripheral blood. The method comprises the following steps of collecting the mononuclear cell of the peripheral blood, first putting the mononuclear cell into a culture bottle enveloped by a CD16 monoclonal antibody, culturing the mononuclear cell, and adding IL-2, IL-15, OK432 and inactivated autoserum into a culture medium; subsequently, transferring the cell into a culture bottle which is not enveloped by the CD16 monoclonal antibody, culturing the cell, adding the IL-2, the IL-15 and the inactivated autoserum into the culture medium; afterwards, transferring the cell into a culture bag, culturing the cell, and adding IL-4 into the culture medium on the last third day, and continuously culturing the cell, so as to obtain abundant NK cells. According to the method, components of animal serum and a tumor cell are not contained; therefore, the safety and the reliability of the NK cells are improved to a great extent; the method has a favorable application prospect.

The invention relates to a method for efficiently amplifying natural killer cells from peripheral blood in a high purity manner. The method comprises the following steps: collecting 100ml-200ml of peripheral blood, separating mononuclear cell, putting the mononuclear cell into a culture bottle coated with CD16 monoclonal antibody and CD226 monoclonal antibody for culturing, and activating the cell by virtue of IFN-gamma; after the activation is carried out for 24 hours, adding IL-2, IL-15, IL-21, OK432, CD2 monoclonal antibody and inactivated autoserum into a culture medium; transferring the cell into a culture bottle which is not coated with CD16 monoclonal antibody and CD226 monoclonal antibody for culturing, and supplementing liquid to the cell once in each 2-3 days; and finally transferring the cell into a culture bag for culturing, and collecting NK cell in the 21th day. According to the method, no any heterologous serum component or trophoblast cell is used, so that the safety and reliability of the prepared NK cell are guaranteed; and the method has good clinical application values.

The invention relates to an in-vitro pure culture method for natural killer cells. The method includes the following steps that peripheral blood is collected, single karyocyte is separated, magnetic beads combined with specific antibodies are adopted to remove the NK cells, and NK cell populations are obtained; the NK cell populations are put into a culture bottle coated with multiple antibodies,and are subjected to activation culture with a complete medium containing IL-2, IL-7, IL-15, anti-CD16, OK432 and inactivated autologous serum, and inactivated NK cell populations are obtained; the inactivated NK cell populations are put into a culture bottle free of antibody coating and continue to carry out multiplication culture, and the natural killer cells are obtained. Any heterologous serumingredients and trophoblast cells are not used in the in-vitro pure culture method, and high amplification efficiency can be wholly obtained; meanwhile, the purity of the amplified NK cells reaches 90% or above, in-vitro efficient pure culture is achieved, and the clinical application requirement is conveniently met.

The invention relates to a preparation method used for high efficiency amplification of natural killer (NK) cells. The preparation method comprises following steps: peripheral blood is collected, andsingle karyocytes are separated; the karyocytes are introduced into a culture bottle treated via coating with CD16 monoclonal antibody and HER2 monoclonal antibody for culturing; IL-2, IL-15, IL-21, and inactivated autologous serum are introduced into a culture medium at the same time; the cells are transformed into a culture bottle free of coating treatment, liquid supply is carried out every 2 to 3 days; and at last the obtained NK cells are collected. According to the preparation method, no heterogenous serum or trophoblastic cell is adopted, operation process is simple, high NK cell yieldis achieved, and the clinical application value is high.

The invention relates to a method for in vitro multiplication culture of NK cells. The method comprises coating a culture bottle with heparin sodium, CD16 and human immunoglobulin, collecting peripheral blood, inactivating plasma, separating mononuclear cells, using a serum-free medium (containing autologous plasma, IL-2, IL-15 and IL-18) to culture multiplication cells, harvesting and detecting the cells and the like. Trophoblast cells or magnetic beads are not used for sorting and purifying, and the method has simple operation, high safety, good amplification effect and high clinical application value.

It is intended to provide a method for producing an NK cell-enriched blood product, the method being less invasive and capable of conveniently and rapidly growing NK cells, etc. in blood collected from an organism. The NK cells in blood are stimulated with NK cell growth-stimulating factors comprising an anti-CD16 antibody, OK432, a bisphosphonate derivative or a salt thereof, or a hydrate thereof, and a cytokine. Then, the blood is cultured at a physiological cell temperature to produce an NK cell-enriched blood product.

Provided is a method for preparing natural killer cell with high efficiency using irradiated peripheral blood mononuclear cells, more particularly to a method for proliferating highly activated NK cells using a combination of irradiated peripheral blood mononuclear cells (PBMCs) and a CD16 antibody and an anti-cancer cell therapeutic composition containing the natural killer cells (NK cells) prepared thereby. Further provided is a method for large-scale proliferation of activated NK cells with high efficiency using a combination of irradiated peripheral blood mononuclear cells (PBMCs) and a CD16 antibody without the use of cancer cells or genetically modified feeder cells having safety issues as feeder cells. The highly purified and highly cytotoxic NK cells proliferated in large quantities can be used as an active ingredient of a cancer immunotherapeutic composition.

The invention belongs to the technical field of cells, and particularly relates to a cell culture solution and cell culture method and application thereof. The cell culture solution comprises a first culture solution and a second culture solution, the first culture solution contains cell factors such as OKT3, CD16, IL-2, IL-15 and 4-1BBL, and the second culture solution contains cell factors such as IL-2, IL-15 and 4-1BBL. The invention further provides the cell culture method. The method comprises the steps that cells are inoculated into the first culture solution for culture, and then the first culture solution is supplemented on the next day; the second culture solution is supplemented when culture is conducted on the ninth day for continuous culture, and the second culture solution is supplemented on the next day. According to the cell culture solution and cell culture method and application thereof, the cell factor combination and culture technological process is optimized, the culture period is shortened, only 14-21 days are needed for culture, and the purity of NK cells obtained through culture reaches 70% or above; the NK cells do not need to be purified, the steps are simple and convenient, and the operability is high.

Disclosed is a medium composition for culturing self-activated lymphocytes, which contains anti-CD3 antibody and anti-CD16 antibody in addition to interleukin 2 (IL-2), interleukin 12 (IL-12) and interleukin 18 (IL-18) in a medium, and thus can efficiently proliferate and activate NK cells, T cells and NKT cells and, at the same time, can significantly increase the ratio of NK cells in lymphocytes so as to provide immunocytes having excellent effects on the treatment of various kinds of malignant tumors, and a method for culturing self-activated lymphocytes using the medium composition.

The invention provides a method for preparing efficient clinical-level CD 56<+> group immune cell. The method comprises the following steps: a) separating single karyocyte from peripheral blood or umbilical cord blood; b) inoculating the single karyocyte in the step a) to an anti-CD16 antibody-coated culture dish; c) adding rabbit-anti-human thymic cell immune globulin, Lifein and animal origin-free cytokine in the culture dish for inducing and stimulating the single karyocyte inoculated in the step b); d) supplementing the culture dish and the animal origin-free cytokine according to the cells counting results every 2-3 days; and e) obtaining the CD 56<+> group immune cell. The method can reduce the cost, accords with the requirement of clinical-level preparation, and has good killing activity in vitro and vivo.

It is intended to provide a method for producing an NK cell-enriched blood preparation, which is low invasive and is capable of conveniently and rapidly growing NK cells, etc. in blood collected from an organism. The NK cells in blood are stimulated with NK cell growth-stimulating factors comprising an anti-CD16 antibody, OK432, an anti-CD3 antibody, and a cytokine. Then, the blood is cultured at a physiological cell temperature to produce an NK cell-enriched blood preparation.

The invention discloses a detection method of human peripheral blood lymphocytes. The preparation method comprises the following steps of: taking human anticoagulation peripheral blood; adding anti-human CD3, CD4, CD8 and CD45 monoclonal antibodies and anti-human CD3, CD56, CD16 and CD45 monoclonal antibodies, then adding a hemolysin working solution containing a fluorescent probe to respectivelyobtain the percentage and absolute count value of a human peripheral blood T lymphocyte subset and an NK lymphocyte subset and achieve the detection of human peripheral blood lymphocytes. The method has the advantages of simplicity and convenience in operation, long sample stable preservation time, high accuracy, multi-index output and the like.

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.