78 results about "Feeder Cell" patented technology

The invention relates to the technical field of biology, in particular to a Treg cell amplification culture medium and a preparation method of Treg cells. According to the method for in-vitro amplification of the regulatory T cells (Treg) from peripheral blood, the amplification efficiency and purity of the Treg cells are improved, the culture cost is reduced, the culture period is shortened, and the problem of trophoblast cell residues in subsequent clinical application is solved.

Several embodiments disclosed herein relate to methods and compositions for enhanced expansion of NK cells in culture, through the repeated co-culturing of NK cells with feeder cells and the selective use of stimulatory interleukins. In several embodiments, the methods utilize one or more soluble interleukins as culture media supplements at one or more time points during expansion of the NK cell, or other immune cell, which results in a highly expanded and highly cytotoxic population of cells, for use in, for example allogeneic cellular immunotherapy.

Described herein are serum-free and feeder-free methods and compositions for the tunable differentiation and production of CD4+ (single positive) T cells and CD8+ (single positive) T cells, by adjusting T cell receptor (TCR) stimulation and notch activation (or notch signaling activation, stimulation or inducement). In some other aspects the invention provides a method and in vitro niche and cell culture media that results in enhancing CD4+ (SP) T cell generation, including naive mature CD4+ single positive T cells from double positive (DP) CD4+8+cells or CD8+(SP) T cells by TCR stimulation while having low or no notch signaling activation, stimulation or inducement. In yet other aspects, the invention provides a method, niche and culture media for producing cells derived from CD4+ (SP) T cells, including Treg cells.

This application provides a method, device, and storage medium for establishing a feeder link. The method includes: determining to provide a feeder service to a gateway station and broadcasting a system message about the feeder cell; and, after establishing a radio resource control (RRC) connection with the gateway station, configuring a feeder data radio bearer (DRB) for the gateway station. This method establishes a feeder link.

A method for generating or expanding a population of natural killer cells (NK cells) via using modified K562 feeder cells having expression of factors capable of increasing activation and proliferation of natural killer cells, wherein the factors capable of increasing activation and proliferation of NK cells include: (A) a co-stimulatory molecule comprises CD80 molecules, and 4-1BBL molecules; and (B) a cytokine, specifically membrane-bound IL-21 (mbIL-21).

The invention discloses construction of a Guangdong small-ear spotted pig spermatogonial stem cell in-vitro culture system and screening of surface markers, and belongs to the technical field of cell culture.The technical scheme includes that testis are broken firstly, then enzyme digestion, density gradient centrifugal purification and differential adherent purification are conducted, and purified spermatogonial stem cells are obtained; inoculating the obtained spermatogonial stem cells to feeder layer cells and culturing in a culture medium; the culture medium is a DMEM culture medium added with FBS and growth factors; the mass fraction of the FBS in the DMEM culture medium is 8-12%, an enzyme digestion method, a density gradient centrifugation method and a differential adhesion method are combined for application, the Guangdong small-ear spotted pig spermatogonia stem cells with certain purity are obtained, cells enriched and purified in vitro can proliferate massively, and an in-vitro culture system of the Guangdong small-ear spotted pig spermatogonia stem cells is established.

Provided are methods for expanding tumor-infiltrating lymphocytes (TILs), which include co-culturing an initial cell population containing TILs with first feeder cells to obtain a first expanded cell population; and then co-culturing the first expanded cell population with second feeder cells to obtain an expanded TIL population. The methods can quickly produce a large number of TILs from a small tumor sample.

The present invention provides a method for producing neural cells or a neural tissue, including the following steps (1)-(3):(1) a first step of culturing pluripotent stem cells in the absence of feeder cells and in a medium containing 1) a TGFβ family signal transduction pathway inhibiting substance and / or a Sonic hedgehog signal transduction pathway activating substance, and 2) a factor for maintaining undifferentiated state,(2) a second step of culturing the cells obtained in the first step in suspension to form a cell aggregate, and(3) a third step of culturing the aggregate obtained in the second step in suspension in the presence or absence of a differentiation-inducing factor to obtain an aggregate containing neural cells or a neural tissue.

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.

The object of this invention is to provide a method of producing a nerve bundle including efficiently extending axons of neural cells. As a solution to accomplish this end, neural cells are cultivated in the presence of feeder cells including at least one type of cells selected from vascular component cells and perivascular cells.

The utility model discloses a feeder cell preparation operating table for a cataract mouse model, which comprises an operating table provided with a sliding hole and a mounting rod locked in the sliding hole through a nut, and further comprises a mounting piece elastically arranged on the mounting rod, the same spring piece is jointly installed at the center positions of the pair of rotating rods, one side of the spring piece is fixedly connected with the installation part, and the end of each rotating rod is sleeved with the same rubber belt so that the spring piece can release elastic potential energy to drive the pair of rotating rods to do circular motion around the rotating shaft to the bottom end to expand and tighten the rubber belt. Through cooperation of the mounting part, the mounting rod, the rotating rod, the spring piece and the rubber belt, the multi-purpose mouse cell extraction operation table is formed, the application range of the operation table is expanded, the operation accuracy is improved, meanwhile, injuries caused by mouse operation are reduced, the structural design is compact and reasonable, operation is convenient and fast, postoperative cleaning and disinfection are easy, and the operation table is worthy of popularization and application. And the practicability is high.

The present invention relates to the application of a dynamically cross-linked hydrogel. Specifically, the dynamically cross-linked hydrogel is formed by mixing gelatin and a cross-linking group-modified cyclodextrin into a hydrogel prepolymer. After embryonic stem cells or immune cells are mixed in, a cross-linking reaction occurs under the action of a cross-linking initiator or ultraviolet light to produce the dynamically cross-linked hydrogel. The hydrogel maintains the stemness of embryonic stem cells in a basal culture medium without the need for feeder cells or differentiation inhibitors, is simple to operate, and is suitable for large-scale culture. The immune cell-loaded hydrogel can be directly used for injection and filling in vivo.

A cell population comprising Corin- and / or Lrtm1-positive cells was produced by the following steps (1) and (2), from which Corin positive and / or Lrtm1 positive cells are collected using a substance that binds to Corin and / or a substance that binds to Lrtm1, and dopaminergic neuron progenitor cells are produced by performing suspension culture of the Corin positive and / or Lrtm1 positive cells in a culture solution containing one or more nutritional factors:(1) a step of performing adhesion culture of pluripotent stem cells in a medium for maintaining undifferentiated state containing a Sonic hedgehog (SHH) signal stimulant, and an undifferentiated state-maintaining factor in the absence of feeder cells but in the presence of an extracellular matrix, and(2) a step of culturing the cell population obtained in the step (1) in a culture solution containing one or more differentiation-inducing factors.

The invention relates to a stem cell culture system purification method based on a double-helix inertial micro-fluidic chip. The double-helix inertial micro-fluidic chip comprises a base and five sequentially connected micro-pipelines fixed on the base, the five sequentially connected micro-pipelines are respectively an inlet micro-pipeline, a first helix micro-pipeline, an S-shaped micro-pipeline, a second helix micro-pipeline and an outlet micro-pipeline, and the first helix micro-pipeline and the second helix micro-pipeline are distributed in a staggered manner. The S-shaped micro-pipeline is positioned in the center of the double-spiral channel formed by the first spiral micro-pipeline and the second spiral micro-pipeline; the front end of the outlet micro-pipeline is divided into a first stage and a second stage, the width of the second stage is larger than that of the first stage, and the tail end of the outlet micro-pipeline is divided into a collection outlet micro-pipeline and a waste outlet micro-pipeline. According to the method, the stem cells are sorted and purified from a stem cell-feeder layer cell co-culture system in a high-flux and label-free manner.

The method for producing a cell aggregate including glial progenitor cells according to the present invention comprises:(1) a step of subjecting pluripotent stem cells to suspension culture in an embryoid-body-forming culture medium containing one or more SMAD signaling inhibitors and one or more Wnt signaling activators in the absence of feeder cells for 5 days to 10 days, to form a cell aggregate;(2) a step of subjecting the cell aggregate obtained in (1) to suspension culture in an embryoid-body-forming culture medium containing retinoic acid;(3) a step of subjecting the cell aggregate obtained in (2) to suspension culture in an embryoid-body-forming culture medium or neuron-and-glia-proliferating culture medium containing retinoic acid and one or more SHH signaling activators; and(4) a step of subjecting the cell aggregate obtained in (3) to suspension culture in a neuron-and-glia-proliferating culture medium containing no retinoic acid and one or more SHH signaling activators.

The present invention relates to a method for inducing and expanding natural killer cells derived from peripheral blood mononuclear cells, which comprises co-culturing, as feeder cells, irradiated Jurkat cells and irradiated Epstein-Barr virus transformed lymphocyte continuous line (EBV-LCL) cells in the presence of cytokines, along with peripheral blood mononuclear cells. According to the present invention, a large quantity of natural killer cells can be induced and proliferated from a small quantity of peripheral blood mononuclear cells even without the use of high-cost equipment or various kinds of expensive cytokines, thereby making it possible to significantly improve the efficiency and efficacy of the prevention and treatment of cancer using the natural killer cells.

The application provides a tumor infiltrating lymphocyte culture medium and application thereof. The culture medium adopted by the application is composed of a basic culture medium, a serum substitute, a low-concentration interleukin, a stimulating factor and an antibiotic and the like. The application discloses a method for preparing high stemness tumor infiltrating lymphocytes (TIL) in vitro under serum-free and feeder-free conditions. Through optimized two-step culture (primary culture for 7-14 days + expansion culture for 7-14 days) and activation stimulation, high stemness TIL products with a CD3+ active cell ratio of greater than or equal to 95% and a Tcm ratio of greater than or equal to 70% can be obtained, and the total expansion multiple reaches 500-1000 times. The method of the application avoids the safety risk brought by feeder cells, reduces IL-2 related side effects, shortens the preparation period to 14-28 days, significantly improves the stem cell characteristics and sustained killing ability of TIL cells, and meets the quality control requirements of clinical-grade cell treatment products.

Disclosed herein are method of producing NK cells that include one or more heterologous nucleic acids. The methods include culturing a population of isolated NK cells in the presence of one or more cytokines to produce a population of activated NK cells. The population of activated NK cells are transduced with a viral vector comprising the one or more heterologous nucleic acids, for example by contacting the activated NK cells with viral particles including the viral vector. The resulting transduced NK cells are then cultured in the presence of one or more cytokines, and optionally in the presence of irradiated feeder cells, to produce a population of expanded transduced NK cells. Also disclosed are methods of treating a subject with a disorder (such as a tumor or hyperproliferative disorder) by administering to the subject NK cells produced by the methods described herein.

The present application relates to a kind of tumor infiltrating lymphocyte (TIL) cell expansion culture process. Specifically, the present application provides a kind of TIL cell expansion method, comprising: in expansion culture medium, preliminary culture TIL cell is carried out fast expansion culture. In the TIL cell expansion method of the present application, it is not dependent on activation antibody CD3 antibody, high-dose IL-2 and feeder cell, that is, the TIL cell number hundreds of times expansion can be realized in short time, the demand of TIL cell quantity in TIL cell therapy clinically can be satisfied, and it has broad application prospect in the field of TIL cell treatment.

An improved method of culturing cells for cell therapy applications that includes growing desired cells in the presence of antigen-presenting cells and / or feeder cells and with medium volume to surface area ratio of up to 1 ml / cm2 if the growth surface is not comprised of gas permeable material and up to 2 ml / cm2 if the growth surface is comprised of gas permeable material. The desired cells are at a surface density of less than 0.5×106 cells / cm2 at the onset of a production cycle, and the surface density of the desired cells plus the surface density of the antigen presenting cells and / or feeder cells are at least about 1.25×105 cells / cm2.

The present invention provides a product comprising plated human hepatocytes on a surface and at least some of the plated hepatocytes are in one or more hepatocyte clusters on feeder cells, which are attached to the surface. A method of preparing plated human hepatocytes is also provided. The preparation method comprises applying human hepatocytes to a surface in the presence of feeder cells, co-culturing the applied hepatocytes with the feeder cells, and forming one or more hepatocyte clusters by the co-cultured hepatocytes on the feeder cells, which are attached to the surface. The plated hepatocytes may be used for various purposes, including the preparation of a hepatitis B virus (HBV) infected hepatocyte culture model and drug testing.

The invention relates to monoclonal antibody production and isolation, and particularly, although not exclusively, to a novel feeder cell line for culturing a monoclonal antibody-producing B cell. The invention also extends to the use of the feeder cell line in culturing a monoclonal antibody-producing B cell, and isolating the B cell from the cell culture media. The invention further extends to methods for culturing and isolating a monoclonal antibody-producing B cell, as well as a method for isolating a monoclonal antibody.

Disclosed is a method for in vitro expansion of tumor-infiltrating lymphocytes. Developed are a new method for isolating and enriching TIL cells in tumor tissues and an optimized culture method for TIL cells independent of feeder cells. 1010 grade or more of TIL cells having an anti-tumor function can be obtained. By means of cell phenotype analysis, compared with TILs expanded by the traditional two-step method, the cultured TIL cells have a higher proportion of memory T cells and a lower proportion of exhausted T cells and regulatory T cells.

The invention discloses a CAR-NK cell of a targeted death receptor 5 as well as a preparation method and application of the CAR-NK cell. The preparation method comprises the following steps: constructing a virus for expressing CAR of targeted DR5; sorting NK cells by adopting a negative selection method; the method comprises the following steps: adopting an NK cell amplification reagent purchased from Yicosai, adding 1-10% of Super Grow purchased from Daraceae into an NK culture medium, and culturing NK cells; and after culturing for 5-9 days, adding a virus for expressing the CAR of the targeted DR5 and a virus transduction enhancer, culturing at 37 DEG C in 5% CO2, and after the cell density is increased to 1-3 * 10 < 6 > cells / mL, carrying out passage to obtain the target DR5. According to the amplification process, the culture medium which is definite in component, free of serum and free of animal source components is adopted, feeder cells and pure cell factors are not used for stimulating and culturing the NK cells from human peripheral blood, large-scale and long-term amplification of the high-purity CAR-NK cells can be achieved, and the amplification process can be used for allogeneic retransfusion.

The present application relates to the technical field of reproductive biology, and specifically discloses a method for culturing mammalian ovary tissue in vitro based on dynamic regulation of gas-liquid interface. In view of the problem that oxygen supply and nutrient supply cannot be optimized and dynamically regulated in the existing in vitro culture of ovary, the method comprises the following steps: preparing an agar block with culture holes; controlling the liquid level of the culture solution to be flush with the outer edge of the agar block and not higher than the upper surface, so that the culture holes are initially liquid-free; placing the mammalian ovary in the holes and directly exposing it to the gas phase environment in the initial stage; during the culture process, the culture solution gradually enters the holes through capillary action, so that the ovary automatically changes to a semi-submerged state, and dynamic changes of the gas-liquid interface are realized. The present application does not require feeder cells or complex scaffolds, and is simple to operate. Through the synergistic effect of initial gas phase exposure and subsequent semi-submerged state, the internal hypoxia of the ovary is significantly improved, cell apoptosis is reduced, and follicle assembly is promoted, so the present application can be used for follicle development mechanism research and reproductive toxicity evaluation.

The present invention relates to an in vitro method for culturing one or more antibody-expressing cells. The method comprises culturing one or more antibody-expressing cells obtained from peripheral blood in the presence of IL-2, IL-21, and non-cell surface-presented CD40 stimulants and in the absence of feeder cells. Furthermore, this specification provides a method for producing antibodies, including the step of culturing one or more antibody-expressing cells according to the method of the present invention, novel CD40 stimulants and their use, and cell culture media.

Provided is a serum-free, xeno-free, feeder-free method for selectively activating and expanding human γδ T cells to achieve sufficient cell numbers for adoptive cell therapy while retaining their robust intrinsic antitumor activity comprising: (a) contacting the γδ T cells with an isolated antibody or antigen-binding fragment that agonizes the γδ T cell receptor; (b) culturing the γδ T cells in cell culture medium comprising a cytokine selected from IL-2, IL-7, IL-12, IL-15, IL-18, IL-21, IL-22, IL-27, and TGFβ, and combinations thereof; wherein the cell culture medium further comprises a serum substitute In some embodiments, the γδ T cells are obtained from induced pluripotent stem cells (iPSCs) and / or peripheral blood mononuclear cells (PBMCs). In further embodiments, the γδ T cells express the Vγ9Vδ2 T cell receptor. The iPSC-derived γδ T cells or the PBMC-derived γδ T cells may be engineered to express a chimeric antigen receptor (CAR).

The present disclosure provides methods of expanding immune cells ex vivo or in vitro comprising culturing the immune cells in a medium comprising a feeder cell replacement, e.g., a PCS, comprising a surface-exposed CD3 agonist. In some aspects, the immune cells comprise T cells, TILs, NK cells, Tregs, or any combination thereof. In some aspects, a population of expanded TILs are administered to a subject in need thereof.

The present disclosure relates to methods and compositions for reprogramming cells to a pluripotent state. In particular, it relates to an integration- and feeder cell-free method for reprogramming primary human fibroblast cells to induced pluripotent stem cells (iPSCs).

The invention discloses a dynamic culture amplification method of NK cells or engineered derivatives thereof, and belongs to the technical field of biology. The method comprises the following steps: sorting peripheral blood to obtain NK cells; mixing the culture medium with K562 feeder cells according to a first proportion, and carrying out first co-culture activation and amplification; optionally, carrying out CAR (chimeric antigen receptor) gene transduction; mixing the amplified NK cells or the transduced cells with K562 feeder cells according to a second proportion, and carrying out secondary co-culture activation and continuous amplification under a dynamic culture condition; and finally, the NK cells or the CAR-NK cells are obtained. The method breaks through the limitation of traditional standing co-culture, realizes efficient and stable amplification of NK / CAR-NK cells under dynamic conditions, has the advantages of high amplification efficiency, high cell viability and purity, simplified operation and easy large-scale amplification, and provides a new technical scheme for preparation of immune cell therapy products.