59 results about "Endoderm cell" patented technology

The invention belongs to the technical field of organoid culture, and particularly relates to a co-differentiated vascularized liver organoid and a construction method thereof. Aiming at the problems that the existing liver organ vascularization mostly adopts a co-culture mode, the operation process is complicated, the cell source heterogeneity is relatively high, the randomness is strong, the uniformity is poor and the like, the invention provides the construction method of the co-differentiated vascularized liver organ. The preparation method comprises the following steps: sequentially inducing and culturing directional endoderm cells, posterior preintestinal cells and hepatic endoderm cells, adding an S1 culture medium containing VEGF to induce and culture for 1-3 days, then adding an S2 culture medium containing VEGF, BMP4 and FGF2 to induce and culture for 3-5 days, and then adding an S3 culture medium containing VEGF to induce and culture for 14-20 days to obtain the co-differentiated vascularized liver organ. According to the method disclosed by the invention, the operation process is simplified, the experiment efficiency is improved, the liver organ which is rich in blood vessel network, high in maturity and of a blood vessel-bile duct structure is obtained, and a foundation is laid for large-scale production and clinical application.

Provided are: a method for producing pancreatic endoderm cells, which comprises a step for culturing pancreatic endoderm cells in a culture medium containing a ROCK inhibitor and KGF and / or EGF; and pancreatic endoderm cells produced by the method.

The invention relates to a cellulose-based hydrogel prepared based on a cellulose copolymer, a cellulose derivative copolymer or a blend thereof, ultramicro fibers of cellulose form a net structure, and the cellulose-based hydrogel has a highly-ordered three-dimensional micro-nano structure, is better in transparency and is more beneficial to observation of a cell state. The cellulose-based hydrogel is suitable for culture of 3D cell culture tissues and organoids, the stem cells cultured by the cellulose-based hydrogel can form cluster culture stem cells and organoids, the operation is simple, compact fused monolayer endoderm cells are formed, non-directional differentiation of the stem cells can be inhibited, and the formed organoids are thick in vesicle wall and are in a solid shape. And the obtained product is a differentiated mature organ and can be used for high-throughput drug screening, drug toxicity testing and regenerative medicine.

The invention belongs to the technical field of regenerative medicine, and particularly relates to a method for generating vascular endothelial cells by inducing pluripotent stem cells to differentiate in vitro. The method comprises the following steps: carrying out first culture on pluripotent stem cells in a differential medium I to obtain cells A; carrying out second culture on the cell A in a differential culture medium II, and differentiating and directionally forming an endoderm cell B; performing third culture on the endoderm cells B in a differential culture medium III to obtain differentiated cells C; carrying out fourth culture on the differentiated cells C in a differentiation culture medium IV, and differentiating to form differentiated cells D; carrying out fifth culture on the differentiated cells D in a differential culture medium V, and differentiating to obtain vascular endothelial cells; wherein the differential culture media I-V do not contain activin A, VEGF (vascular endothelial growth factor) and BMP4 (bone morphogenetic protein 4). The method can improve the differentiation efficiency of the IPS cells to the vascular endothelial cells, and has a wide prospect in the cell therapy industry.

Described are small molecule cocktails that enable de novo derivation and long-term culture of bovine extraembryonic endoderm cells (bXENs). Methods of using the small molecule cocktails to generate bXENs are also described. Methods of using the bXENs to form blastocyst models or to maintain the stemness of bovine ESCs and prevent them from differentiation are also described.

The invention provides a method for 3D differentiation of pluripotent stem cells into definitive endoderm cells by using a bioreactor, a differentiation medium and a differentiation factor A / B are also included, and the medium and the differentiation factor A / B comprise the following components: an MCDB131 medium, 2.44 mM glucose, 29.28 mM sodium bicarbonate, 2% of HSA, 1% of GlutaMAX, 250 [mu] M Vitamin C, 0.002% of ITS-X, 1% w / v of Polyethylene glycol (PEG), 1 mg / ml of Heparin Sodium salt, 100 ng / mL of Activin A and 3 [mu] M of CHIR99021. The method uses the bioreactor to differentiate the pluripotent stem cells into the stereotyped endoderm cells in a 3D suspension manner, is simple to operate and high in differentiation efficiency, provides technical support for scientific research and industrial large-scale production, and has a good application prospect.

The invention relates to the technical field of biology, in particular to a method for efficiently obtaining human definitive endoderm cells in vitro, which comprises the following steps: S1, inoculating human pluripotent stem cells into matrigel, and performing maintenance culture for 16-24 hours by using a stem cell maintenance culture medium; s2, from Day-2 to Day-1, absorbing and abandoning the stem cell maintenance culture medium, adding a pretreatment culture medium for culture, and changing the pretreatment culture medium once every day; the pretreatment culture medium is a stem cell maintenance culture medium added with dimethyl sulfoxide; s3, carrying out Day0 to Day2, absorbing and abandoning the pretreatment culture medium, adding a definitive endoderm differential culture medium, carrying out induced culture, and changing the definitive endoderm differential culture medium once a day; the definitive endoderm differential culture medium is an RPMI1640 culture medium added with fetal calf serum and activin A; s4, Day3, obtaining the required human definitive endoderm cells, and solving the problems that the existing definitive endoderm differentiation efficiency and cell population purity are limited and are influenced by batch-to-batch variation.

To provide a method for manufacturing a hepatoblast, a hepatoblast, and a method for manufacturing a hepatocyte.SOLUTION: A method for manufacturing a hepatoblast includes a step of culturing an iPS cell-derived entodermal cell serum-free medium containing FGF2, HGF, oncostatin M and dexamethasone and differentiating the cell into a hepatoblast. As one aspect, the medium further contains nicotinamide, or the serum-free medium is serum-free differentiation medium, or an iPS cell is derived from a human, or a culture period of the iPS cell-derived entodermal cell is 5 days or more and less than 10 days.SELECTED DRAWING: Figure 2-1

Provided is a high-function organoid composition. Also provided is a method of producing the organoid composition. The organoid composition is characterized by including: an organoid including a cell cluster of endodermal cells; and an amphiphilic block polymer having a hydrophilic block chain having a sarcosine unit and a hydrophobic block chain having a lactic acid unit. The organoid composition may be produced by culturing the endodermal cells in a culture system containing the amphiphilic block polymer. The organoid composition thus obtained can evaluate pharmacokinetics and drug toxicity, and is hence suitable for drug screening.

The application belongs to the technical field of biology, and discloses a kit for inducing stem cells to differentiate into megakaryocytes and / or platelets and application thereof. The kit can induce stem cells to differentiate into megakaryocytes and / or platelets without introducing exogenous cells and exogenous genes. The kit can highly simulate the generation process of megakaryocytes and platelets in vivo, and can induce stem cells into mesendoderm cells, hematopoietic endothelial cells and megakaryocyte-erythroid progenitor cells in sequence. The megakaryocyte-erythroid progenitor cells further develop into megakaryocytes with a multiploid characteristic, and finally produce stem cell-derived platelets with similar functions to natural platelets. The stem cells have the potential for unlimited proliferation, and can be massively expanded in vitro, which provides an unlimited cell source for the in vitro preparation of platelets.

Disclosed are cell encapsulation devices and methods for transplanting cells, such as pancreatic endoderm cells, into a host. In some examples, a cell encapsulation can comprise a lumen configured to receive cells therein, a cell-excluding membrane, where the lumen is internal to the cell-excluding membrane, and a non-woven fabric layer external to the cell-excluding membrane, where the non-woven fabric layer and the cell-excluding membrane comprise perforations. The device can further comprise a woven mesh external to the non-woven fabric layer, where the non-woven fabric layer provides protection to the cell-excluding membrane from direct contact with the woven mesh.

The present invention belongs to the field of organoid culture technology, and specifically relates to a co-differentiated vascularized pancreatic islet organoid and a method for constructing the same. In view of the problems that the existing pancreatic islet organoid vascularization mostly adopts the co-culture method, which has a long operation process, is difficult, and has randomness and poor uniformity, the present invention provides a method for constructing a co-differentiated vascularized pancreatic islet organoid, which adopts human induced pluripotent stem cells hiPSCs, and sequentially induces and cultures them into directed endoderm cells, primitive intestinal tube cells and posterior foregut cells. In the subsequent induced differentiation, a culture medium containing VEGF, BMP4 and FGF2 is added for culture to obtain co-differentiated vascularized pancreatic islet organoid. For the first time, the present invention directs the differentiation of iPS cells to produce vascularized pancreatic islet organoids containing β, α, δ cells and vascular endothelial cells without adding exogenous endothelial cells. It is more mature than non-vascularized pancreatic islet organoids and has a stronger insulin secretion function, providing new ideas and experimental basis for achieving functional cure of diabetes.

The present application relates to the technical field of biology, and particularly discloses a method for culturing and obtaining foregut endoderm cell spheroids and application thereof. The method for culturing and obtaining foregut endoderm cell spheroids of the present application obtains foregut endoderm cell spheroids by inducing differentiation of stem cells on a culture surface with a PDA-matrigel coating. The culture method of the present application can ensure the differentiation efficiency of DE and AFE adherent cells and spheroids, promote the generation of AFE cell spheroids and the separation from the adherent cell layer, reduce the edge rolling of AFE adherent cells, prolong the production time of AFE cell spheroids, significantly improve the yield of AFE cell spheroids, and provide a basis for subsequent differentiation and culture of lung organoids.

Embodiments of the instant disclosure relate to novel compositions and methods for generating thymic cells. In some embodiments, thymic cells can be differentiated from pluripotent stem cells (PSC), anterior primitive streak (APS) cells, definitive endoderm (DE) cells, anterior foregut endoderm (AFE) cells, pharyngeal endoderm (PE) cells, ventral pharyngeal endoderm (VPE) cells, and third pharyngeal pouch endoderm (TPPE) cells using the compositions and methods disclosed herein. In certain embodiments, thymic cells generated by composition, systems and methods disclosed herein can be used to treat a health condition.

Methods of producing synthetic embryos are provided. Accordingly, a method of producing a synthetic embryo is provided, comprising inducing expression of a factor in a subset of primitive pluripotent stem cells (PSC) that induces differentiation into trophoderm cells to obtain trophoderm originating cells; inducing expression of factors which are induced to differentiate into extraembryonic primitive endoderm cells in the second subgroup of the primitive state PSC to obtain extraembryonic primitive endoderm originating state cells; and mixing the trophoblast ectoderm originating state cells and the extraembryonic primitive endoderm originating state cells with primitive state PSC under conditions that allow the formation of aggregated cells. Articles, mixtures, and aggregates of cells and methods of using the same are also provided.

Methods and compositions for generating human pancreatic beta cells and progenitor cells thereof from human pluripotent stem cells using chemically defined media are disclosed. The method can preliminarily generate FOXA2 + HNFlb + dorsal anterior intestinal endoderm cells (DFECs), and according to the provided method, the cells can be further differentiated into human PTF1A + PDX1 + pancreatic progenitor cells (PPCs) and human INS + PDX + pancreatic beta cells (PBCs). The invention also provides a culture medium, an isolated cell population and a kit.

In a first embodiment, a method for producing fetal and / or neonatal hepatocyte-like cells (fnHLCs) is disclosed. The method includes producing a definitive endoderm cell population by a stage 1 process, producing a hepatoblast-like progenitor cell population from the definitive endoderm cell population by a stage 2 process, and producing a fetal and / or neonatal hepatocyte-like cell population from the hepatoblast-like progenitor cell population by a stage 3 process. In another embodiment a method for screening a target is disclosed. The method includes co-culturing and incubating the target with a fetal and / or neonatal hepatocyte-like cell (fnHLC), retrieving a sample of a fnHLC after the co-culturing and incubating and determining one or more characteristics of the fnHLC based on analysis of the sample of the fnHLC.

The present invention discloses the application of fatty acid synthesis metabolic pathway inhibitors in the differentiation of definitive endoderm cells, and also discloses a culture medium and a culture method for the in vitro directed differentiation of mammalian pluripotent stem cells into definitive endoderm cells. The culture medium comprises a basal medium and an inhibitor of the fatty acid synthesis metabolic pathway; the culture method is to culture mammalian pluripotent stem cells using the above-mentioned culture medium to enable their directed differentiation into definitive endoderm cells. By adding the fatty acid synthesis metabolic pathway inhibitor Firsocostat or C75 to the basal medium, the present invention improves the efficiency of differentiating mammalian pluripotent stem cells into definitive endoderm cells. The differentiation method provided by the present invention has good repeatability and low cost, and improves the efficiency of differentiating mammalian pluripotent stem cells into definitive endoderm. This provides low-cost and highly efficient research materials for studying early embryonic development, organ transplantation, and drug screening, etc.

This method for inducing mesendodermal cells comprises a step for increasing the amount of intercellular zinc finger protein 384 (ZNF384) existing in pluripotent stem cells.

The present invention discloses a mesenchymal stem cell derived from a definitive endoderm cell, a differentiation method and an application thereof, and relates to the technical field of mesenchymal stem cells. The differentiation method comprises: inducing human pluripotent stem cells to differentiate into mesendoderm cells by culturing in a first differentiation medium containing a WNT pathway activator and Activin A for 1-2 days; inducing the formation of definitive endoderm cells (DE) by culturing in a second differentiation medium containing no WNT pathway activator and Activin A for 2-8 days; inducing the acquisition of midgut, hindgut and foregut cell fate by culturing in a MSC generation medium for 2-4 days; inducing the generation of a cell population containing mesenchymal stem cells by culturing in a MSC generation medium for 8-10 days; performing digestion and passage 2-3 times to obtain mesenchymal stem cells (DE-MSCs) derived from definitive endoderm cells. It can secrete anti-inflammatory factors under in vitro stimulation and has the potential to treat colitis.

To provide single hormonal insulin producing cells differentiated from pluripotent stem cells, and an in vitro method for differentiation.SOLUTION: The present invention provides an in vitro differentiated population of pancreatic endoderm cells obtained from the stepwise differentiation of pluripotent cells, wherein cells at each step of differentiation are cultured in medium comprising 5 mM to 20 mM glucose.SELECTED DRAWING: None

Embodiments of the present disclosure relate to novel compositions and methods for generating thymic cells. In certain embodiments, thymus cells can be differentiated from pluripotent stem cells (PSC), anterior primitive stripe (APS) cells, definitive endoderm (DE) cells, anterior intestinal endoderm (AFE) cells, pharyngeal endoderm (PE) cells, abdominal pharyngeal endoderm (VPE) cells, and third pharyngeal sac endoderm (TPPE) cells using the compositions and methods disclosed herein. In certain embodiments, thymic cells generated by the compositions, systems, and methods disclosed herein can be used to treat health conditions.

The present invention discloses the application of sodium acetate in improving the efficiency of differentiating mammalian pluripotent stem cells into definitive endoderm cells, and also discloses a culture medium and a culture method for differentiating mammalian pluripotent stem cells into definitive endoderm cells. The culture medium contains a basal medium and sodium acetate; the method is to culture mammalian pluripotent stem cells using the above-mentioned culture medium to direct their differentiation into definitive endoderm cells. By adding sodium acetate to the basal medium, the present invention improves the efficiency of differentiating mammalian pluripotent stem cells into definitive endoderm cells. The differentiation method provided by the present invention has good repeatability and low cost, improves the efficiency of differentiating mammalian pluripotent stem cells into definitive endoderm, and thus can be widely applied to the research of cell fate determination and organ repair, etc.

The application provides an iPS-driven nasopharyngeal epithelial organoid and a preparation method and application thereof. The preparation method comprises the following steps: differentiating humanized iPS cells into endoderm cells; wrapping the endoderm cells with Matrigel and then placing the endoderm cells in DMEM / F-12 culture medium containing Dorsomorphin and SB431542 for culture to obtain foregut organoids; culturing the foregut organoids in DMEM / F-12 culture medium containing FGF10 protein, CHIR-99021, retinoic acid and BMP4 protein to obtain airway epithelial progenitor cell organoids; and culturing the airway epithelial progenitor cell organoids in DMEM / F-12 culture medium containing FGF10 protein, retinoic acid and BMP4 protein to obtain nasopharyngeal epithelial organoids. The iPS cells are continuously differentiated in multiple stages by using culture medium containing different inducers, and the nasopharyngeal epithelial organoids are mass-produced.

While studies have been made on a method of selectively inducing differentiation from pluripotent stem cells into enterocyte-like cells, provided is an excellent and high-functionality enterocyte-like cell population that can be stably tested for multi-drug metabolism and permeability, and also provided an efficient method of producing cells. The method of inducing differentiation from pluripotent stem cells into enterocyte-like cells includes the following steps (1) and (2): (1) a step of inducing differentiation from pluripotent stem cells into definitive endoderm cells; and (2) a step of culturing the definitive endoderm cells in a system containing CHIR99021, followed by induction of differentiation into intestinal progenitor cells. A cell population of enterocyte-like cells is more effectively obtained by seeding and culturing enterocyte-like cells produced by the method of inducing differentiation, in a base material for organoid production or a base material for two-dimensional culture.

To provide a method for producing in vitro functional beta-cells (functional pancreatic beta-cells) and populations of cells resulting from the differentiation of pluripotent stem cells.SOLUTION: A method of differentiating pancreatic endoderm cells into functional beta cells expressing PDX1, NKX6.1, MAFA, UCN3, and SLC2A1, comprises culturing immature pancreatic beta cells in a medium supplemented with (a) an effective amount of T3 and (b) an effective amount of 3-deazaneplanocin A (DEZA), thereby producing functional beta cells expressing insulin.SELECTED DRAWING: None

The present invention discloses the application of an AMPK activator in improving the efficiency of differentiating mammalian pluripotent stem cells into definitive endoderm cells, and also discloses a culture medium and a culture method for the in vitro directed differentiation of mammalian pluripotent stem cells into definitive endoderm cells. The culture medium contains a basal medium and an AMPK activator EX229; the culture method is to culture mammalian pluripotent stem cells with the above-mentioned culture medium to enable their directed differentiation into definitive endoderm cells. By adding the AMPK activator EX229 to the basal medium, the present invention improves the efficiency of differentiating mammalian pluripotent stem cells into definitive endoderm cells. The differentiation method provided by the present invention has good repeatability and low cost, and improves the efficiency of differentiating mammalian pluripotent stem cells into definitive endoderm, which provides a more efficient tool for the research on early embryonic development and disease treatment.

To provide a device and method for grafting a cell such as a pancreatic endoderm cell to a host.SOLUTION: A cell delivery device includes a nonwoven fabric outside a cell excluding membrane, and can drill the nonwoven fabric and / or the cell excluding membrane. Treatment of a host by immune suppression reagent is required in order to suppress allograft rejection by drilling of the device, and does not impair maturation or function of a grafted pancreatic endoderm cell.SELECTED DRAWING: Figure 1A

To provide a cell structure that includes a lumen, an endodermal cell, an ectodermal cell and a mesodermal cell, and also includes a macrophage, and a method for manufacturing the structure.SOLUTION: A method for manufacturing a cell structure that includes a lumen, an endodermal cell, an ectodermal cell and a mesodermal cell, and also includes a macrophage, includes the steps of: injecting a monocyte into the cell structure including a lumen, an endodermal cell, an ectodermal cell and a mesodermal cell; and maturing the monocyte to a macrophage in a cell structure into which the monocyte has been injected.SELECTED DRAWING: Figure 3A

To provide compositions, systems, kits, and methods for generating dorsal foregut endoderm (DFE) cells by contacting pluripotent stem cells with a retinoic acid signaling pathway agonist and a bone morphogenetic (BMP) pathway inhibitor without exposing the stem cells to a transforming growth factor beta (TGFβ) pathway agonist.SOLUTION: In certain embodiments, the DFE cells are contacted with a retinoic acid signaling pathway agonist and a FGFR pathway inhibitor to generate pancreatic endoderm (PE) cells (e.g., with dorsal identity). In other embodiments, the PE cells are contacted with an ALK5 inhibitor and a Notch inhibitor to generate endocrine cells (e.g., insulin expressing endocrine cells). In certain embodiments, compositions, systems, kits, and methods are provided for generating anterior domain endoderm (ADE) cells.SELECTED DRAWING: None