13 results about "Feeder Layer" patented technology

This invention discloses a method for preparing NK cells, the NK cells obtained thereby, and their applications. The method includes: sorting NK cells from peripheral blood of an allogeneic donor; culturing them in a serum-free medium containing CD3 monoclonal antibody, CD16 antibody, and IL-2, without the need for a feeder layer; preferably, IL-21 is added for amplification on days 3-5 of culture, and the cells are cultured for 20-40 days. This method yields non-genetically modified NK cells, among which CD56... bright The NK cells comprise >90% of the cell line and highly express a variety of activating and functional molecules. These NK cells effectively clear Aβ oligomers and senescent cells, and alleviate neuroinflammation. In treatment, intracranial injection achieves equivalent or better efficacy with only about 1 / 10th the dose administered intravenously, significantly clearing Aβ deposits in the brains of AD model mice and improving pathology.

This invention relates to the field of bovine embryonic stem cell culture technology, and particularly to a long-term culture system for morphological bovine embryonic stem cells completely without a feeder layer and its applications. After trying various culture methods, this invention establishes a feeder-free bovine morphological embryonic stem cell line (FF-bESCs). Bovine morphological embryonic stem cells cultured under the feeder-free conditions provided by this invention can maintain stable proliferation capacity and pluripotency over a long period. The bovine morphological embryonic stem cells provided by this invention can undergo gene editing and can be effectively induced to differentiate into primordial germ cell-like cells (PGCLCs). This invention lays an important foundation for the standardized and large-scale culture of bovine embryonic stem cells under feeder-free conditions, and for their application in stem cell breeding and other fields.

The application belongs to the technical field of cell biology, and particularly relates to a feeder-free and serum-free bovine embryonic stem cell culture medium, a culture system and a culture method. The bovine embryonic stem cell culture medium comprises a conditioned medium and an additive. The additive is IWR-1 and activin A. The concentrations of IWR-1 and activin A in the conditioned medium are 2.5-5 muM and 20 ng / mL-25 ng / mL respectively. The culture system comprises the bovine embryonic stem cell culture medium, 0.1%-0.2% gelatin in mass concentration and a subculture medium. The 0.1%-0.2% gelatin in mass concentration is used to provide an attachment matrix for the bovine embryonic stem cell. The subculture medium is prepared by adding Y27632 to the bovine embryonic stem cell culture medium. The application reduces the cost of extracellular matrix, and the feeder-free bovine embryonic stem cell has high single cell subculture efficiency and is convenient for gene editing operation.

A method of differentiating nonhuman primate induced pluripotent stem cells (iPSCs) to oligodendrocytes is provided. The method may include providing a modified vector comprising a piggyBac vector backbone that expresses one or more differentiation factor genes, including SOX10, OLIG2, and NKX6-2. The piggyBac vector backbone may comprise one or more terminal inverted repeats and one or more transposase recognition sites configured to interact with a piggyBac transposase to mediate genomic integration at TTAA or noncanonical target sequences. The vector may further include a selectable marker and a promoter for regulated expression. The modified vector may be introduced into nonhuman primate iPSCs, for example from Macaca fascicularis, by electroporation and used in a feeder-free differentiation workflow. The resulting differentiated cells may be identified by expression of oligodendrocyte-associated markers including O4, O1, PDGFRA, MOG, GALC, CNPase, and myelin basic protein (MBP).

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.

The invention provides a method for inducing efficient amplification of NK cells through the synergistic effect of exosome and immunosuppressor, and belongs to the technical field of cell culture, and the method comprises the following steps: 1) extracting and purifying stem cell exosome; the stem cells are human umbilical cord mesenchymal stem cells; 2) separating CBMC cells and plasma from the umbilical cord blood, inactivating the plasma, activating platelets, incubating and separating to obtain super plasma; 3, CBMC cells are inoculated and cultured in a low-oxygen incubator for 10-16 h, then the exosomes in the step 1 are added, culture continues to be conducted for 10-16 h, the CBMC cells are transferred to a normal oxygen environment, culture does not exceed 4 h, a culture medium and super plasma are supplemented, culture continues to be conducted, and amplified NK cells are obtained.According to the technical scheme, in-vitro culture of the high-activity NK cells can be efficiently amplified and maintained without a feeding layer is achieved.

The present application relates to a method of deriving a human embryonic stem cell (hESC) line under feeder-free and heterologous conditions.

The invention relates to the technical field of biotechnology, and discloses a construction method of a bovine originating embryonic stem cell line, which comprises the following steps: (1) obtaining in vitro fertilized bovine early blastocyst; (2) trophoblast cells of the blastocyst are removed, an inner cell cluster is inoculated into a culture system containing a feeder layer, and the culture system is a basic culture medium added with FGF2 and WNT pathway inhibitors; and (3) culturing until an inner cell cluster forms a cell set drop, and carrying out digestion passage to obtain the stably proliferated bovine originated embryonic stem cell line. According to the construction method and the application of the bovine embryonic stem cell line, a specific initial induction combination of 'FGF2 + WNT inhibitor IWR1' is adopted, and a cell mass in an early blastocyst is used for establishing the line, so that the bovine embryonic stem cell line which is uniform in form, can be stably passaged for more than 26 generations and still keeps typical embryonic characteristics is successfully obtained; the problems of unstable state and easy differentiation of the bovine stem cell line in the prior art are solved.

The present disclosure relates generally to novel methods and compositions for using engineered reprogramming factor(s) for the creation of induced pluripotent stem cells (iPSCs) through a kinetically controlled process. Specifically, this disclosure relates to establishing combinations of reprogramming factors, including fusions between conventional reprogramming factors with transactivation domains, optimized for reprogramming various types of cells. More specifically, the exemplary methods disclosed herein can be used for creating induced pluripotent stem cells from various mammalian cell types, including human fibroblasts. Exemplary methods of feeder-free derivation of human induced pluripotent stem cells using synthetic messenger RNA are also disclosed.

The present application relates to the field of cell biology, specifically will be by adding phosphoantigen (HMBPP), IL-2 and other factors, specific activation of gamma delta T cells, and Daudi lymphoma cell feeder layer co-culture, promote the expansion of gamma delta T cells, at the same time, promote the memory phenotype and survival of gamma delta T cells, and improve the cytotoxicity and function of gamma delta T cells. The method simultaneously induces peripheral blood mononuclear cells to become gamma delta T cells and activates gamma delta T cells, enhances immune signal transduction, and significantly improves the activation efficiency and expansion capacity. The results show that the proliferation of gamma delta T cells is 742.62 times in 14 days, which is significantly higher than that of conventional CIK culture; CD3 + cell ratio reaches 90.15%, which is better than 75.29% of CIK. Thus, a short cycle, high yield, and good purity gamma delta T cell preparation process is formed, which embodies the creativity of multi-factor synergy and feeder layer co-culture, and brings significant beneficial effects.

The present invention aims to provide a method for producing iPS cell-derived keratinocytes that can induce keratinocytes that highly express keratinocyte markers in a feeder-free and serum-free environment. [Solution] A method for producing iPS cell-derived keratinocytes, comprising an induction step of inducing iPS cells into keratinocytes using one or more induction media in a feeder-free and serum-free environment, wherein for at least a portion of the induction step, a medium containing fibroblast growth factor 7 (FGF7) is used as the induction medium.

The invention relates to the technical field of biology, in particular to a combined culture medium of chicken embryo primordial germ cells (Primordial Germ Cells, PGCs) and a cell culture method of the combined culture medium, calcium ion-free DMEM is used as a basis, various supplements, nucleoside, sodium pyruvate, amino acid, vitamins, additives, trace components, three antibodies, serum and specific cell factors are added, filtration sterilization is performed after osmotic pressure is adjusted, and the combined culture medium and the cell culture method of the combined culture medium are used for culturing the chicken embryo primordial germ cells (Primordial Germ Cells). The cell factors are added before use to adapt to the culture of the related PGCs. The method does not need a feeding layer, simplifies the operation, avoids exogenous pollution, improves the purity and survival state of the PGCs, promotes proliferation, has no sex preference, is adaptive to different sources of PGCs, is stable and reliable, and has a wide application prospect.