99 results about "Cell soma" patented technology

The present invention relates to the discovery of a novel molecular pathway involved in long-term hyperexcitability of sensory neurons, which, in higher animals, is associated with persistent pain. It is based on the discovery that, following injury to an axon of a neuron, an increase in nitric oxide synthase activity results in increased nitric oxide production, which, in turn, activates guanylyl cyclase, thereby increasing levels of cGMP. Increased cGMP results in activation of protein kinase G (“PKG”), which then is retrogradely transported along the axon to the neuron cell body, where it phosphorylates MAPKerk.

An apparatus and method for purification and assay of neurites is useful for separation and analyses of extension organelles and/or protrusion of cells for purification, production, observation, and quantification of neurites in the neurobiology field. The present invention provides a pore-sized controlled porous filter membrane which outspace side surface is coated with a cell adhesion layer to form an adhesion surface, and combines the neuronal cells with the porous filter membrane in an aqueous environment under conditions in which outgrown neurites from cell bodies of the neuronal cells are attracted to and grow on the adhesion surface, wherein the outgrown neurites of the neuronal cells pass through pores provided in the porous filter membrane to the adhesion surface while each of the pores has a size smaller than the cell bodies of the neuronal cells so as to prevent the cell bodies of the neuronal cells passing through the pores and remaining on an opposing side surface of the porous filter membrane.

The invention discloses a composite microcapsule model for use in in-vitro cell coculture. The model is a sphere-in-sphere composite microcapsule model consisting of an inner sodium alginate gel microsphere and an outer sodium alginate gel microsphere, which is formed by using natural polysaccharide sodium alginate as a substrate material, dripping the polysaccharide sodium alginate into multivalent cation solution and performing gelatinization treatment twice, wherein the two gel microspheres may contain two different kinds of cells. The model can be used for the study on in-vitro coculture of various cells and overcomes the drawback that the interaction between cells of the same kinds and the interaction between cells of different kinds cannot be controlled, the drawback that different kinds of cells are difficult to separate in late-stage study and the drawback that cell dependency exists of the conventional coculture model; and the model can simulate cell-cell interaction, cell-extracellular matrix interaction and cell-soluble factor interaction and provide theoretical and technical support for in-vitro construction of a tumor model, optimization of in-vitro culture mode of cells and tissue engineering construction of various tissue and organ analogues or equivalents.

The invention provides morphology-based neuron recognizing and analyzing method. The method comprises the following steps: acquiring microscopic image data of nerve cells; preprocessing the microscopic image data to obtain the noise-removed image; performing threshold separating for the noise-removed image to obtain the background-removed neure image; extracting single neure from the neure image; extracting basic parameters of single neure, wherein the basic parameters include a framework, a cell body, area of the cell body, quantity of axon or dendron, and length of axon or dendron. With the adoption of the method, the neure in the microscopic image can be automatically recognized and analyzed; in addition, the recognition is fast, and the ideal effect is realized; moreover, the shape of the neure can be visually observed; the effective neure basic parameters can be output; therefore, the labor intensity of a researcher reading a picture can be greatly reduced; meanwhile, subsequent data analysis can be performed conveniently.

The invention provides a method for preparing umbilical cord mesenchymal stem cells and aims to obtain umbilical cord tissues. The method comprises the steps of conducting digestive treatment on the umbilical cord tissues with Whartons jelly digestive juice, and collecting the digestive juice, wherein the Whartons jelly digestive juice comprises hyaluronidase and a chondroitin sulfate ABC enzyme which separate and extract the umbilical cord mesenchymal stem cells from the digestive juice. By means of the method for preparing the umbilical cord mesenchymal stem cells, the hyaluronidase and the chondroitin sulfate ABC enzyme are adopted for digesting Whartons jelly in a joint mode in the enzymic digestion process, the umbilical cord mesenchymal stem cells with the best cell quality can be obtained on the whole, the influence of long-time digestion on cell bodies by a pancreatic enzyme and collagenase is avoided, and the obtaining rate of the umbilical cord mesenchymal stem cells which are finally obtained through the preparation is higher; the obtained stem cells have higher cell viability and differentiation potential.

The invention discloses the technical field of NK cell amplification. The invention relates to a method for efficiently amplifying NK cells in vitro by a factor method. The method comprises the following steps: carrying out cell separation, cell inoculation and activation, liquid exchange and cell amplification; according to the invention, a CD16 monoclonal antibody and a NKP46 monoclonal antibodyare matched for use, the OK432 is used, and the IL-15 and the hydrocortisone are matched for use; the method is characterized by directly culturing PBMCs by utilizing cured Anti-CD16mAb and NKP46 monoclonal antibodies; the culture bottle does not need to be pre-coated; OK432 is used as a biological adjuvant; NK cells are amplified in vitro under the combined action of hydrocortisone, IL-2 and IL-15; and hydrocortisone is an immunosuppressive agent. When used with IL-15 at the same time; hydrocortisone can be used for promoting proliferation of IL-15 activated NK cells; according to the present invention, the efficient NK cell in vitro amplification method is created, the CD3-CD16+/CD56+ cell expression rate of the NK cell prepared through the method is up to 60% or more, and after 15-18 days of culture, the NK cell can be amplified by 1000 or more.

The invention relates to the field of biotechnology and medical science, in particular to a fusion protein for NKT (natural killer T) cell culture, an encoding gene and an application. The fusion protein is used for preparing NKT cells which are high in proliferating activity and killing activity. The fusion protein ICAM1-FN (intercellular adhesion molecule-1-fibronectin) can effectively stimulate proliferation of lymphocytes in the culture process of the NKT cells, the proportion of CD3/CD56 double-positive cells is improved, so that the in-vitro amplification efficiency of peripheral blood mononuclear cells is twice that of a traditional method, the proportion of CD3/CD56 double-positive cells is 2-10 times that of the traditional method, tumor killing activity of the NKT cells is enhanced, and the tumor killing activity of the prepared method of the NKT cells is improved by 15%-35% as compared with the traditional method.

The invention provides an NK cell in-vitro amplification system. The NK cell in-vitro amplification system comprises an activation culture medium for cell activation and a proliferation culture medium for cell proliferation, the activating culture medium is composed of an activating agent and a basic culture medium, and the activating agent is a combination of solid-phase anti-CD52 and IL-2; the proliferation culture medium is composed of a proliferation agent and a basic culture medium, and the proliferation agent is IL-2. Compared with the prior art, the NK cell in-vitro amplification system disclosed by the invention has the following technical effects: 1) the culture medium contains the immobilized anti-CD52, and the NK cells can be efficiently stimulated to enter an activated and proliferated state by combining the use of IL-2, so that a large number of activated NK cells can be easily obtained; moreover, no trophoblasts are used in the culture process, and the prepared NK cells can be used for clinical research and treatment; and 2) the NK cell culture method is simplified, few NK cell activation components are used, few material resources and manpower are used, the production cost of NK cell preparation is obviously reduced, and the method is particularly suitable for large-scale production.

The invention relates to a construction method and an application of a M2-type tumor-associated macrophage model. In the invention, an immortalized mononuclear cell (for example, a U937 monocytic series) is induced into a tumor-associated macrophage in vitro environment under stimulation of PMA and IL-4, IL-10 and TGF[beta] growth factor; and on the basis of the mentioned before, biological functions and target therapy value of the tumor-associated macrophage in tumor progression are researched in a manner of in-vivo and in-vitro co-cultivation method of the tumor-associated macrophage and tumor cells. The invention provides important tools and experimental measures for tumor immunology fundamental research and therapy measures based on the tumor-associated macrophage.

The invention relates to an in vitro amplification method of self-specific T cells, a T cell system prepared by the method, pharmaceutical use of the cell system, and a component monitoring method of the cell system. The in vitro amplification method comprises the steps of: (1) separating peripheral blood mononuclear cells from peripheral blood of a patient; (2) adding corresponding T cell stimulating factors as needed, adding or not adding corresponding cell growth factors, and amplifying T cells by cultivating the peripheral blood mononuclear cells separated in the step (1) in a corresponding culture medium. According to the invention, polyclonal immunocompetent cells with various functionalities can be prepared according to different clinical needs by adopting the amplification method of the invention in a GMP environment, and the prepared polyclonal immunocompetent cells can be used in immune system regeneration and immunotherapy of patients.

The invention discloses a polypeptide and application of the polypeptide in NK cell culture and preparation of an NK cell culture medium. The amino acid sequence of the polypeptide is as follows: KGNP(XX) TDHQVFWMWMWMLGEAAYD, wherein XX represents D or G. The polypeptide provided by the invention not only can promote in-vitro proliferation of NK cells, but also can improve the lethality of the NKcells, so that the polypeptide can be used for promoting in-vitro proliferation of the NK cells and enhancing the tumor killing activity of the NK cells, and can be used for preparing a cell culturemedium capable of promoting in-vitro proliferation of the NK cells and enhancing the tumor killing activity of the NK cells.

The invention provides a targeting CLL1 chimeric antigen receptor and an application thereof. The targeting CLL1 chimeric antigen receptor comprises an antigen binding structural domain, a hinge region, a transmembrane structural domain and a signal transduction structural domain, and the antigen binding structural domain is an anti-CLL1 antibody. The anti-CLL1 antibody is adopted as an antigen binding structural domain to construct a chimeric antigen receptor molecule, the targeted CLL1 chimeric antigen receptor has a specific targeting effect on CLL1 positive tumor cells, the in-vivo and in-vitro killing effect of immune cells expressing the targeted CLL1 chimeric antigen receptor is remarkable, a large number of cell factors IFN-gamma are secreted after co-culture with the CLL1 positive tumor cells, and the product has a specific removal effect on CLL1 positive tumor cells.

The present invention belongs to the technical field of medicines and specifically relates to an application of ulinastatin in preparations of anti-nasopharyngeal carcinoma metastasis medicines. Through cell in-vitro tests, the ulinastatin is found to be capable of inhibiting metastasis of nasopharyngeal carcinoma cells, does not inhibit growth of the nasopharyngeal carcinoma cells, and is less toxic and more conducive to rehabilitation of patients with nasopharyngeal carcinoma.

The invention discloses a polypeptide and application in promoting proliferation of NK cells and improving killing ability of the NK cells to tumor cells. The amino acid sequence of the polypeptide isKGNPPTDHQVFWM (XX) MLGEAAYD, wherein XX represents W or G. The polypeptide provided by the invention can not only promote the proliferation of the NK cells in vitro, but also increase the killing ability of the NK cells, so that the polypeptide can be used to promote the proliferation of NK cells in vitro and enhance tumoricidal activity of the NK cells, and be used to prepare a cell culture medium which can promote the proliferation of NK cells in vitro and enhance the tumoricidal activity of the NK cells.

A method of manipulating and/or investigating cellular bodies (9) is provided. The method comprises the steps of: providing a sample holder (3) comprising a holding space (5) for holding a fluid medium (11); providing a sample (7) comprising one or more cellular bodies (9) in a fluid medium (11) in the holding space (5); generating an acoustic wave in the holding space exerting a force (F) on thesample (7) in the holding space (5). The method further comprises providing the holding space (5) with a functionalised wall surface portion (17) to be contacted by the sample (7) and the sample (7) is in contact with the functionalised wall surface portion (17) during at least part of the step of application of the acoustic wave. A system and a sample holder (3) are also provided.

Disclosed below is a device comprising a base 112, polymer walls 105, comb fingers 104, groove/ridge architecture 107, metal film 108, comb buses 103, electrical ground electrode 109, and polymer well 101 for electrical and mechanical stimulation as well as for measuring of contractile force and conduction velocity of the cellular sheet/cluster 102 in response to mechanical and electrical stimulation of the cell source as it grows into a multilayer cellular sheet on the device. This allows cell sheets to be cultured and conditioned to be compatible with the patient's cardiac environment in vitro, prior to sheet release and implantation. Major innovative elements of this device include: real-time data for rich understanding of engineered tissues as they are grown, ability to expose engineered tissues to patient-derived stimuli (specifically localized electrical stimuli, mechanical stimuli, and micro-architecture), and the option to implant after characterization.

The invention provides a method for establishing an analysis module for assessing neurological functions. The method comprises the following steps: acquiring culture cells which include a plurality of fluorescent labels by virtue of a micro-fluorescent imaging system so as to conduct image analysis, wherein the culture cells include nerve cells and non-nerve cells; in accordance with area size of cell nucleuses and fluorescence intensity, selecting the nerve cells showing axon and dendrite fluorescent labels, eliminating non-nerve cells, and calculating the area sizes of the nerve cell bodies as well as axon and dendrite lengths and the number of branches, so that axon and dendrite growth situations of the nerve cells are determined; and in accordance with axon and dendrite fluorescent labeling ranges, calculating the number of synaptic puncta fluorescent labels on the nerve cells, so that the growth situation of the nerve cells is determined, and the neurological functions are assessed. The method disclosed by the invention can further serve as a drug screening platform, so as to rapidly detect whether drugs have a protective or neurotoxic effect on the nerve cells or not.

Through a genetic engineering technology, a LMP2A-targeted chimeric antigen receptor extracellular region LMP2A scFv segment is constructed by using a high affinity full human anti-LMP2A Fab as a template. A third generation LMP2A-targeted chimeric antigen receptor is constructed by recombination of the LMP2A-targeted chimeric antigen receptor(LMP2A CAR) extracellular region LMP2A scFv segment andsignal peptide CD8alpha, transmembrane region CD8TM and intracellular regions CD28, CD137 and CD3 zeta. The prepared LMP2A-targeted chimeric antigen receptor is recombined into a linearized lentiviral vector by overlay PCR to construct an anti-LMP2A CAR lentiviral expression vector, a LMP2A CAR virus solution is prepared, and T cells are infected to obtain LMP2A CAR-T cells. In vitro and in vivotests validate the LMP2A CAR-T cells have specific targeted killing effects on LMP2A positive nasopharyngeal carcinoma cells, and provide a new therapeutic idea and means for the treatment of nasopharyngeal carcinoma.

The invention discloses an in-vitro amplification optimization method for regulatory T cells derived from cryopreserved umbilical cord blood. The method comprises the following steps of: resuscitating the cryopreserved umbilical cord blood, and then performing centrifugal washing to obtain total karyocytes; then inoculating the total karyocytes into a culture dish pre-coated with an Anti-human CD3 antibody, and adding an induction medium for induction culture; and performing CD4 magnetic bead sorting and purification on the eighth day of culture to obtain CD4+ regulatory T cells, and then continuing second-stage induction culture amplification to finally obtain high-purity regulatory T cells. The method provided by the invention can obtain the high-purity regulatory T cells from the cryopreserved umbilical cord blood through in-vitro amplification optimization. The method is also suitable for in-vitro amplification optimization culture of the regulatory T cells of fresh umbilical cord blood.

The invention provides a cell in-vitro culture device. The cell in-vitro culture device comprises a peripheral wall (10) and a base (20); the peripheral wall (10) is of a hollow structure with openings at two ends; the base (20) comprises a bottom plate (21); one end of the peripheral wall (10) is fixed on the bottom plate (21); a connecting port (22) is formed in the bottom plate (21); the connecting port (22) is communicated with the hollow structure; a mounting surface (23) is formed on one side, deviating from the peripheral wall (10), of the bottom plate (21); the mounting surface (23) isused for mounting a cell culture membrane (50); and, when the cell culture membrane (50) is mounted on the mounting surface (23), the peripheral wall (10), the base (20) and the cell culture membrane(50) define an open container for placing a cell solution. The cell in-vitro culture device provided by the invention can meet the experimental requirements of different types of biological materials, and is convenient for more accurate experiments of some membrane materials.

The invention discloses a cell culture medium for improving DC cell activity and a culture method, and belongs to the technical field of biomedicine. The culture medium comprises an RPMI 1640 culture medium as well as comprises pomalidomide. According to the method, when the DC cells are cultured in vitro, a proper amount of pomalidomide is added, so that the DC cell activity and maturity in healthy subjects and multiple myeloma patients are remarkably improved, indicating a potential as a DC adjuvant. The cell culture medium of the present invention can be applied to DC-based treatment strategies such as DC vaccine and DC cell therapies.

The invention discloses a cell culture medium additive, a cell culture medium and a cell in-vitro amplification method. The cell culture medium additive comprises 10 to 40 mg/mL of human serum albumin, 0.5 to 10 ng/mL of EGF, 0.5 to 10 ng/mL of KGF, 0.5 to 10 ng/mL of FGF-10, 0.1 to 1.0 [mu] g/mL of hydrocortisone, 1 to 10 [mu] g/mL of insulin and 1 to 10 [mu] g/mL of transferrin. In addition, the invention also discloses an application of the cell culture medium additive in cell culture. According to the technical scheme provided by the invention, the in-vitro amplification efficiency of the human protodermal keratinocytes can be improved, and the obtained cells can support the construction of a three-dimensional recombinant human skin model with intact structures and functions.

The invention discloses a primary regulatory T cell in-vitro amplification system. The amplification system is prepared by adding anti-CD3/28 antibody, interleukin-2. Rapa and transforming growth factor-beta 1 into a basic culture medium, wherein the final concentration of the anti-CD3/28 antibody is 2 microgram/ml, the final concentration of the interleukin-2 is 1000U/ml, the final concentrationof the Rapa is 100-200 nanogram/ml, and the final concentration of the transforming growth factor-beta 1 is 4-8 nanogram/ml. Compared with an amplification system which uses the Rapa alone, the Rapa and transforming growth factor-beta 1 combined amplification system is good in stability and high in Foxp3+regulatory T cell purity.