49 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.

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 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.

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.

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.

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 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.

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.

The invention provides an in vitro method for producing an isolated mammalian primitive retina stem cell (pRSC), comprising: (a) culturing an isolated embryonic stem cell (ESC) from a mammal in a cell culture medium that is free of feeder cells, feeder layer conditioned medium, or serum to produce and grow a culture of the isolated ESC; and (b) contacting the culture of the isolated ESC so grown with one or more of a Wnt or TGF-beta / BMP signaling inhibitor to differentiate the isolated ESC of (a) into a primitive retina stem cell, thereby producing an isolated mammalian pRSC.

This invention relates to a modified feeder cell and a method for culturing natural killer (NK) cells, aiming to enhance the proliferation capacity and killing activity of NK cells by using a specific feeder cell.

The invention provides culture platforms, cell media, and methods of differentiating pluripotent cells into hematopoietic cells. The invention further provides pluripotent stem cell-derived hematopoietic cells generated using the culture platforms and methods disclosed herein, which enable feed-free, monolayer culturing and in the absence of EB formation. Specifically, pluripotent stem cell-derived hematopoietic cell of this invention include, and not limited to, iHSC, definitive hemogenic endothelium, hematopoietic multipotent progenitors, T cell progenitors, NK cell progenitors, T cells, NK cells, NKT cells and B cells.

The present invention relates to an in vitro method for cultivating one or more antibody expressing cell(s). The method comprises cultivating one or more antibody expressing cell(s) obtained from peripheral blood in the presence of IL-2, IL-21 and a non-cell surface presented CD40-stimulating agent and in the absence of feeder cells. Moreover, herein provided are methods for producing antibodies comprising the step of cultivating one or more antibody expressing cell(s) according to the method of the invention, a novel CD40-stimulating agent and uses therefrom as well as a cell culture medium.

Provided are an HLA-E modified feeder cell and a method for culturing natural killer cells (NK cells). By means of culturing NK cells using the HLA-E-modified feeder cell, NK cells having a low expression of inhibitory receptor NKG2A and a high expression of activating receptors, such as NKG2C and NKp44, can be obtained, thereby significantly enhancing the killing activity of NK cells against target cells, particularly those with a high HLA-E expression.

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 relates to methods and compositions for inducing differentiation of hematopoietic cells. The present invention provides culture platforms, cell culture media, and methods for differentiating pluripotent cells into hematopoietic cells. The present invention further provides pluripotent stem cell-derived hematopoietic cells generated using the culture platforms and methods disclosed herein that allow for feeder cell-free monolayer culture without EB formation. In particular, the pluripotent stem cell-derived hematopoietic cells of the present invention include, and are not limited to, iHSCs, permanent hemogenic endothelial cells, hematopoietic multipotent progenitor cells, T cell progenitor cells, NK cell progenitor cells, T cells, NK cells, NKT cells, and B cells.

The invention relates to the technical field of immune cell culture, in particular to a co-culture method for large-scale immune cells. Comprising the following steps: S1, performing irradiation treatment on umbilical cord mesenchymal stem cells, and then performing cryopreservation; s2, reviving the umbilical cord mesenchymal stem cells subjected to irradiation treatment, inoculating the umbilical cord mesenchymal stem cells and NK cells into a co-culture bottle, and performing co-culture through a circulating device; the circulating device comprises a circulating pool, a peristaltic pump, a cell filter, a shaking table and an air supply pump; the circulating pool is connected with the co-culture bottle through a liquid inlet pipe and a liquid outlet pipe, peristaltic pumps are respectively arranged on the liquid inlet pipe and the liquid outlet pipe, and the cell filter is connected in series with the liquid outlet pipe; the co-culture bottle is placed on the shaking table, and the air supply pump is connected with the circulating pool through a pipeline. Umbilical cord mesenchymal stem cells are used as trophoblast cells to provide nutrition and proliferation for NK cells, then a circulating device is used to prevent the cells from clustering and aging, and the NK cell proliferation is promoted while the killing ability of the NK cells to tumor cells is improved.

The invention provides an NK cell high amplification method. Specifically, the invention provides a method for preparing the NK cells, and the method comprises the following steps: 1) culturing CD3-mononuclear lymphocytes in a first-stage culture medium in the presence of feeder cells to obtain a first cell population containing the NK cells; 2) adding the first cell population into a second-stage culture medium to obtain a second cell population containing NK cells; and 3) culturing the second cell population in a third-stage culture medium to obtain a third cell population containing the NK cells so as to prepare the NK cells, wherein the first-stage culture medium and the second-stage culture medium contain serum, and the third-stage culture medium does not contain serum.

The invention provides a feeder cell density and motility rate detection kit and a detection method thereof, and belongs to the technical field of cell detection. The invention specifically discloses a method for detecting cell density and / or motility rate, which comprises the following steps: (1) exposing feeder cells to a CFSE dye environment for dyeing; (2) removing the CFSE dye in the environment; (3) culturing the feeder cells under proper conditions; (4) adding absolute counting microspheres into the cultured feeder cells and exposing the feeder cells in a dead cell dye environment for dyeing; and (5) calculating the density or the motility rate of the living cells according to a dyeing result and absolute counting microspheres. The method has good specificity, accuracy, precision and durability, and has good popularization value and market application prospect.

The invention relates to a method for isolating embryonic stem cells (100) from at least one avian embryonic cell, comprising: isolating at least one embryo (101) around the developmental stage of oviposition; suspending embryonic cells (102) obtained from the embryo by dissociation step a) in an animal serum-free base medium supplemented with a combination of growth factors, at least one inhibitor and an animal serum substitute; seeding the embryonic cell suspension (103) on a feeder cell layer; culturing (104) the embryonic cells at least once passaged.