11 results about "NK cell proliferation" patented technology

The invention relates to the technical field of NK cell culture, and discloses an NK culture medium containing hypotaurine and application of the NK culture medium to improvement of NK cell proliferation and killing activity, and the NK culture medium comprises a basic NK cell culture medium and hypotaurine; the basic NK cell culture medium does not contain hypotaurine and comprises amino acid, vitamins, inorganic salt, trace elements, D-glucose and sodium pyruvate. When the basic NK cell culture medium is an RPMI1640 culture medium, the concentration of the hypotaurine is 0.1-1 mM; when the basic NK cell culture medium is an IMDM culture medium, the concentration of the hypotaurine is 0.4-4 mM. According to the scheme, the hypotaurine is added into the basic NK cell culture medium to obtain the NK culture medium, the NK culture medium can increase the number of NK cells when the NK cells are cultured in vitro, and the killing activity of the cultured NK cells on tumor cells is remarkably improved.

The present application relates to the technical field of cell expansion, in particular to a NK cell culture method based on multi-factor combined induction and application. The present application combines CD16, OK432, IL-2, IL-15, IL-18 and NK cell proliferation peptide and other factors to synergistically promote the proliferation of NK cells, so that the in vitro proliferation activity of NK cells is significantly improved, which is conducive to promoting the application of NK cells in clinical treatment.

This invention relates to the field of NK cell culture technology, and discloses an NK culture medium containing taurine and its application in improving NK cell proliferation and killing activity. The medium includes a basal NK cell culture medium and taurine; the basal NK cell culture medium does not contain taurine and includes amino acids, vitamins, inorganic salts, trace elements, D-glucose, and sodium pyruvate. When the basal NK cell culture medium is RPMI_1640 medium, the concentration of taurine is 0.1~1mM; when the basal NK cell culture medium is IMDM medium, the concentration of taurine is 0.4~4mM. This method obtains an NK culture medium by adding taurine to the basal NK cell culture medium. This NK culture medium not only increases the number of NK cells during in vitro culture, but also significantly enhances the killing activity of the cultured NK cells against tumor cells.

The invention discloses a traditional Chinese medicine compound cell culture fluid and application thereof. The traditional Chinese medicine compound cell culture fluid comprises a basic culture medium, and 1 L of the basic culture medium contains 2-10 mg of notoginsenoside R1, 1-5 mg of notoginsenoside Rb1, 1-5 mg of notoginsenoside Rg1, 0.1-1 mg of notoginsenoside Rg3, 0.5-2 mg of notoginsenoside Re, 1-6 [mu] g of shikonin, 5-15 [mu] g of cordycepin and 1-20 mg of basil polysaccharide. In the traditional Chinese medicine compound cell culture fluid, notoginsenoside Rg3 can be combined with shikonin, so that the cytotoxicity can be enhanced, cord blood NK cells are promoted to be differentiated to mature functional phenotypes with high cytotoxic activity, and the killing efficiency of the NK cells on various tumor target cells is remarkably improved; the notoginsenoside Rb1 is combined with the basil polysaccharide, so that the NK cells can be driven to expand and grow, the activity of the NK cells can be maintained, and the cell purity can be improved, so that the persistence of the cell activity can be enhanced, and the tumor killing rate of the NK cells can be improved.

The present application belongs to the field of cell culture. The present application provides a method for inducing, culturing and amplifying human natural killer cells, comprising the following steps: extracting peripheral blood from a healthy person, and separating PBMC; coating a container with an NK activation solution; culturing with an NK cell proliferation medium, inoculating PBMC, and incubating at a constant temperature for 1-6 days; and proliferating with an NK cell amplification medium, and incubating at a constant temperature for 1-10 days. The present application effectively improves the in-vitro proliferation rate of human natural killer cells. Through flow cytometry analysis, the proportion of NK cells with phenotypes of CD3, CD16 and CD56 in the cell population is all above 80%, and the cell phenotype is CD56 + CD3 ‑ NK cell group, and the positive rate is 97.57%. The present application has a simple composition formula, ideal culture effect, and wide application prospect.

The present application provides a kind of interleukin 2 mutant, fusion protein, antibody or conjugate comprising the mutant and pharmaceutical composition comprising the mutant, fusion protein, antibody or conjugate. Compared with wild type interleukin 2, the mutant eliminates the binding with IL-2R alpha, reduces the binding with IL-2R beta gamma dimer, but maintains the stimulation of tumor immune cells, including but not limited to T cells and NK cells proliferation. The IL-2 mutant of the present application eliminates the affinity for high affinity IL-2 receptor, while reducing the affinity for medium affinity IL-2 receptor. The interleukin 2 mutant of the present application is more suitable for fusion protein, antibody conjugate. The mutation site of the interleukin 2 mutant of the present application is very few relative to wild type interleukin 2, so the potential immunogenicity is very low. At the same time, there is no various side effects produced by immunotherapy with natural IL-2.

The invention discloses a trispecific antibody which comprises a first antigen binding region, a second antigen binding region, a compound of IL-15 and a receptor thereof, and an Fc fragment, the first antigen binding region has CD16 binding activity; the second antigen binding region has HLAG binding activity; the compounds of the first antigen binding region and the second antigen binding region IL-15 and the receptors thereof are directly or indirectly connected with the Fc fragment respectively. The trispecific antibody can be combined with HLAG on tumor cells and CD16 and IL-15 receptors on NK cells at the same time, cell synapse is promoted to be formed between the two cells, then the NK cells are activated to kill HLAG positive tumor cells, and NK cell proliferation can be promoted. Therefore, the trispecific antibody provided by the invention can kill HLAG positive tumor cells, and can effectively treat HLAG-mediated related diseases (such as cancers).

The invention relates to the technical field of improved breeding of animal husbandry, and particularly discloses an improved NK cell culture medium and a preparation method thereof. The improved NK cell culture medium comprises a basic culture medium, a supplement and a composite sustained-release body. All the components are mixed according to a specific proportion, and the requirements of NK cell proliferation and function maintenance are met. Wherein the inner layer of the composite sustained-release body takes trehalose and human recombinant serum albumin as protective agents to wrap the interleukin, and the outer layer of the composite sustained-release body is an acrylate prepolymer photocuring polymer containing a photoinitiator, so that the continuous and stable release of the interleukin is realized. Through component optimization and slow release system design, the NK cell culture efficiency and activity are effectively improved, and the method is suitable for NK cell multiplication culture.

The invention relates to the technical field of cell amplification, in particular to an NK cell culture method based on multi-factor combined induction and application. Various factors such as CD16, OK432, IL-2, IL-15, IL-18 and NK cell proliferation promoting peptide are combined to synergistically promote proliferation of the NK cells, so that the in-vitro proliferation activity of the NK cells is obviously improved, and the application of the NK cells in clinical treatment is promoted.

This invention relates to proteins targeting B7H3 and methods of using them. Multispecific B7H3-targeting compounds and methods of using them are provided. These multispecific compounds include bispecific and trispecific targeting proteins. The bispecific targeting protein includes a B7H3 targeting domain and an immune cell binding domain; the trispecific targeting protein includes a B7H3 targeting domain, an immune cell binding domain, and an immune cell activation domain. These methods of use include methods for inducing NK-mediated cell killing, methods for inducing NK cell proliferation in vivo, and methods for treating cancer.

This invention relates to the field of immune cell culture technology, and particularly to a method for in vitro culture of peripheral blood NK cells. The invention involves sequentially performing a first culture and a second culture on in vitro cultured peripheral blood NK cells. In the first culture stage, star anise extract and IL-2 are used. The active components in the star anise extract synergistically interact with IL-2 to induce an increase in NK cell activity. Furthermore, in the second culture stage, epimedium extract is used to culture the NK cells, which improves the NK cell environment and promotes rapid NK cell proliferation. The entire process requires no additional treatment and is simple. Therefore, the technical solution of this invention not only enhances the proliferation capacity of NK cells but also prolongs their original cytotoxic activity.