141 results about "Feeder Cell" patented technology

A high-throughput microfluidics-based fluorescence activated droplet sorting (FADS) system provides activation, sorting, and feeder cell-free expansion of NK cells that are enriched for killing activity. The NK cells are activated and identified based on their ability to kill target cells, such as tumor cells, without prior sensitization. The diversity of endogenous NK cells can be overcome to allow selection of a highly effective population of cells for therapeutic use, such as in the treatment of cancer.

The invention provides a method for enhancing Treg cell functions and application, and belongs to the technical field of cell engineering. According to the method, hUC-MSCs are irradiated by gamma rays to form feeder layer cells, and then the feeder layer cells and Treg cells are co-cultured to obtain the Treg cells with enhanced cell functions. Through the co-culture of the feeder layer cells and the Treg cells, the inhibition ability of the Treg cells on activation and proliferation of immune cells can be enhanced, and the feeder layer cells and the Treg cells can be used as universal cells in clinical researches of autoimmune diseases, graft versus host diseases and the like.

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 a method for differentiating induced pluripotent stem cells into megakaryocytes and platelets, and relates to the technical field of induced pluripotent stem cell directional differentiation, and the method comprises the following steps: inoculating an iPSC single cell into a culture vessel, and carrying out adherent culture through a culture solution; after the cell adheres to the wall and before cloning is not formed, the liquid is changed into a first induction culture liquid, and the cell is induced and differentiated into a mesoderm cell; changing the solution into a second induction culture solution to induce and differentiate the mesoderm cells into hematopoietic endothelial cells; collecting the suspension cells, culturing the suspension cells in an amplification culture solution, and enriching megakaryocytes; inducing megakaryocyte polyploidy through a third induction culture solution; and culturing the polyploidy megakaryocyte through a fourth induction culture solution to promote platelet release. The method for inducing the pluripotent stem cells to differentiate into megakaryocytes and platelets is convenient and rapid, the megakaryocytes and platelets can be obtained through iPSC in a short time, and feeder cells, serum and additional gene editing are not needed; meanwhile, the yield is high.

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.

The present disclosure provides feeder hydrogel particles that can function to support the growth, proliferation, and / or activation of a target cell in culture. The present disclosure also provides methods of culturing target cells with feeder hydrogel particles.

The invention provides a construction method and application of a gene modified K562 cell line, and belongs to the technical field of biological medicines. According to the invention, two viruses CD86-4-1BBL and mbIL-21 are respectively subjected to gene transduction packaging by adopting a third-generation lentivirus four-plasmid packaging system, a K562 cell line is infected by the lentiviruses CD86-4-1BBL and mbIL-21, and a monoclonal cell line capable of stably expressing CD86, 4-1BBL and mbIL-21 proteins is obtained through screening. According to the method, cell lines with the cloning numbers of V5-E9, V10-A5, V5-D6 and V6-D7 are obtained through screening, the four cloning cell lines can serve as feeder cells for stimulating NK cell amplification after being amplified and subjected to irradiation treatment, NK cell amplification can be effectively promoted, the amplification multiple of the NK cells reaches up to 6000-8000, and large-scale preparation and production of the NK cells for treatment are met.

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

The present invention relates to a method for inducing the differentiation of corneal epithelial cells from pluripotent stem cells. More specifically, the present invention relates to a method for autonomously differentiating pluripotent stem cells, such as human iPS cells, into ectodermal cell lineage in a serum-free medium without using feeder cells and inducing the differentiation of the resultant ocular surface ectodermal lineage cells into corneal epithelial cells.

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 producing natural killer cells is disclosed. The method comprises isolating peripheral blood mononuclear cells (PBMCs) from a blood sample; isolating at least one of CD56+ cells and / or CD3− / CD56+ cells from the PBMCs; and co-culturing the at least one of CD56+ cells and / or CD3− / CD56+ cells with a combination of feeder cells in the presence of a cytokine. A composition for treating cancer is also disclosed. The composition comprises the CD56+ natural killer cells produced by the disclosed method and a cytokine.

To provide a method for producing induced pluripotent stem cells (iPS cells) that can initialize somatic cells without using feeder cells or a substrate. For production of iPS cells, the following steps are carried out: I. introducing an initialization gene into a somatic cell; and II. performing initialization and amplification culture of the cell into which the gene has been introduced, in a liquid medium comprising at least one of a protein kinase Cβ (PKCβ) inhibitor and a WNT inhibitor under a suspension culture condition.

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 provides a feeder cell for amplifying NK (Natural Killer) cells as well as a preparation method and application of the feeder cell. The feeder cell is a human cell with over-expression of IL-15, IL-21, 4-1BBL and IL-12p35 on a cell membrane. The human cells are selected from any one of immune cells, immortalized immune cells, lymphoma cells or leukemia cells. When the feeder cells are used for amplifying the NK cells, the amplification efficiency is high; the amplified NK cells are stronger in killing function; the feeder cell has very high clinical application value in amplification of the NK cells and large-scale clinical application of the NK cells.

The invention provides a method for obtaining GiNK cells through novel culture medium amplification, a large number of highly purified human natural killer (NK) cells are obtained by combining IL-15 active fragments, PI3K inhibitors, key receptors targeting the surfaces of the NK cells, antibodies enhancing activation of the NK cells and various cytokines, and the GiNK cells can be used for preparing the human natural killer cells. Expansion from human peripheral blood mononuclear cells (PBMCs) can be achieved without the use of feeder cells. Through culture for 2-3 weeks, the GiNK cells can be amplified by hundreds to thousands of times, and the proliferation efficiency, functional activity and anti-tumor effect of the GiNK cells are remarkably improved. The culture medium provided by the invention not only reduces the use concentration of cell factors, but also solves the problem of function depletion of GiNK cells in in-vitro culture through the synergistic effect of various additives, and provides important technical support for clinical application of the GiNK cells.

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 disclosure provides conditionally immortalized stem cells and cell lines generated therefrom. The conditionally immortalized stem cells and cell lines generated therefrom can be used to prepare immortalized feeder cell lines, tester cell lines, and / or therapeutic cell lines.

The present disclosure provides an engineered population of natural killer cells wherein the NK cells express at least one polypeptide selected from the group consisting of human leukocyte antigen E, human leukocyte antigen F, and human leukocyte antigen G. The NK cell comprises a genetically modified disruption of one or more copies of an endogenous beta-2 microglobulin and a genetically modified disruption of one or more copies of a human leukocyte antigen class II-related gene, the human leukocyte antigen class II-related gene is selected from a protein containing regulatory factor X-related ankyrin, a regulatory factor 5, a regulatory factor X-related protein and a class II trans-activator. Also provided are a method of producing human pluripotent stem cell-derived engineered natural killer cells without feeder cells, and a method of treating a disease or condition in a subject by administering the NK cells to the subject.

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 invention relates to a multiplication culture process of tumor infiltration lymphatic (TIL) cells. Specifically, the invention provides a propagation method of TIL cells, which comprises the step of carrying out rapid propagation culture on preliminarily cultured TIL cells in a propagation culture medium. In the TIL cell expanding culture method, hundreds of times of amplification of the TIL cells can be realized in a short time without depending on an activated antibody CD3, high-dose IL-2 and feeder cells, the requirement of TIL cell therapy on the number of the TIL cells clinically can be met, and the TIL cell expanding culture method has a wide application prospect in the field of TIL cell therapy.

The present invention relates to a method for amplifying an invariant natural killer T (iNKT) population and an application thereof, the method comprising: providing a peripheral blood stem cell sample; culturing the peripheral blood stem cell sample in an amplification culture medium, wherein the amplification culture medium comprises interleukin 2, interleukin 15 and alpha-galactosylceramide; and adjusting the number of the cultured cells, and co-culturing the cultured cells and feeder cells in the amplification medium, the feeder cells being a dendritic cell and a peripheral blood mononuclear cell.

The present invention relates to the production and isolation of monoclonal antibodies, in particular (but not limited to) novel feeder cell lines for culturing B cells that produce monoclonal antibodies. The invention also relates to application of the feeder cell line in culturing B cells generating monoclonal antibodies and in separating the B cells from a cell culture medium. The invention also relates to a method for culturing and isolating B cells producing monoclonal antibodies and a method for isolating monoclonal antibodies.

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 disclosure relates to methods for expanding and increasing the cytotoxic activity of natural killer cells comprising co-culturing, as feeder cells, a population of myeloid leukemia cells engineered to express one or more of membrane-bound IL-21 (mbIL-21) or membrane-bound IL-15 (mbIL-15) in the presence of cytokine support. The present disclosure also relates to a population of acute myeloid leukemia cells engineered to express one or more of membrane-bound IL-21 (mbIL-21) or membrane-bound IL-15 (mbIL-15). The present disclosure also relates to methods of treating cancer employing the step of expanding natural killer cells using feeder cells engineered to express one or more of membrane-bound IL-21 (mbIL-21) or membrane-bound IL-15 (mbIL-15).

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.