31 results about "Endoderm cell" patented technology

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.

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.

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.

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 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 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

The present disclosure provides cell-based compositions for treating diabetes, methods for identifying cells that preferentially differentiate into endoderm cells, and methods for preparing insulin-producing pancreatic cells, as well as related methods of use for treating diseases related to insulin deficiency.

Embodiments of the present disclosure relate to novel compositions and methods for generating thymocytes. In some embodiments, thymocytes can be differentiated from pluripotent stem cells (PSCs), 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, thymocytes generated by the compositions, systems, and methods disclosed herein can be used to treat health conditions.

To provide ventral hindgut organoids for producing bladder organoids with a layered structure of bladder epithelial cell types similar to those in the bladder.SOLUTION: One aspect of the present invention provides a method for producing ventral hindgut organoids, comprising culturing pluripotent stem cells using induction medium A containing activin A and a GSK3β inhibitor to induce differentiation into definitive endoderm cells, and culturing the definitive endoderm cells using induction medium B that contains a fibroblast growth factor and a GSK3β inhibitor and further may contain a bone morphogenetic protein, and then culturing the cells in induction medium B that contains a fibroblast growth factor and a GSK3β inhibitor and further may contain a bone morphogenetic protein, in the presence of an extracellular matrix, to form ventral hindgut organoids.SELECTED DRAWING: None

To provide: processes for differentiating an endodermal cell into a posterior foregut cell, a posterior foregut cell into a hepatic progenitor cell, and / or a hepatic progenitor cell into a hepatocyte-like cell; and obtained cell populations.SOLUTION: In some embodiments, the process can be conducted in the absence of serum. The hepatocyte-like cell population obtained from this process have a detectable CyP3A4 activity and / or express a detectable level of albumin and / or of urea. The process can be designed to increase cellular yield.SELECTED DRAWING: None

Provided herein are 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. 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.

The present invention relates to methods of efficiently generating pancreatic endoderm from human pluripotent stem (PS) cell derived human definitive endoderm. The present invention also relates to pancreatic endoderm cells obtained by the methods of the invention. Finally, the present invention relates to culture medium and composition comprising a RAR antagonist and uses of said RAR antagonist in the induction of pancreatic endoderm cells. The present invention provides a more homogenous and synchronised pancreatic cell population, with increased efficiency.

The invention discloses a method for preparing intestinal organs from human induced pluripotent stem cells, and belongs to the technical field of biomedicine. A related culture differentiation system is optimized, stem cells are efficiently induced to be sequentially differentiated into endoderm cells and midgut and posterior gut cells, and finally intestinal organs with a complete intestinal tissue structure are formed. The method comprises the following key steps: amplifying stem cells by using a special culture medium, taking 3-5 generations of cells, and carrying out synergistic induction on the cells by Activin A and Wnt3a to differentiate into an endoderm; fGF-4, CHIR99021 and Y-27632 are used for promoting the formation of middle and posterior intestine spheres and the maturation of precursor cells; and finally, forming the organoid in a Matrigel three-dimensional scaffold and a compound factor culture medium of EGF (Epidermal Growth Factor), Noggin, RSPO1, N2 and B27. The method can be used for intestinal development mechanism research, disease model construction, drug screening and regenerative medicine, and particularly can be used for establishing a model by utilizing the specific stem cells of the patient in personalized medicine, so that precise drug testing and treatment scheme optimization are realized.

The present invention relates to a method for accelerated and cost-effective generation of insulin-producing beta cells from pluripotent stem cells using a fully defined, small molecule-based differentiation protocol. Further, the present invention relates to a method for obtaining pancreatic endoderm cells from posterior foregut cells. The use of the cells obtained with the method according to the invention is described as well.

The present invention provides: a method for producing pancreatic endoderm cells, the method comprising a step of culturing pancreatic endoderm cells in a culture medium containing a ROCK inhibitor, and KGF and / or EGF; and pancreatic endoderm cells produced by said method.

This invention relates to a method for preparing cardiac organoids derived from human pluripotent stem cells, comprising: Step A: culturing human pluripotent stem cells (hPSCs) to prepare embryonic bodies with a diameter of 100 μm to 130 μm; Step B: culturing the embryonic bodies prepared in Step A to a diameter of 200 μm to 250 μm; Step C: differentiating the embryonic bodies derived from human pluripotent stem cells with a diameter of 200 μm to 250 μm prepared in Step B into constructs containing mesodermal cells and endoderm cells; Step D: differentiating the constructs containing mesodermal cells and endoderm cells differentiated in Step C into constructs containing cardiomyocytes, fibroblasts, and vascular endothelial cells. Self-organized cardiac organoids (cells); and, step E: culturing and maturing the self-organized cardiac organoids differentiated through step D, wherein the cardiac organoids that have completed the maturation process through step E have cardiomyocytes, fibroblasts, and endothelial cells forming a self-organized structure with a cell ratio of 55 to 65:15 to 30:15.

A method of culturing wild-type hepatitis A virus (wtHAV) using induced differentiated pluripotent stem cell-derived hepatocytes. Specifically, iPSCs were used to differentiate into definitive endodermal (DE) cell, and then the DE cells were differentiated into four stages of hepatocyte progenitor (HpSC), hepatoblasts (HB), immature hepatocytes (immHep), and derived hepatocytes (dHep), and each stage cells were inoculated with wtHAV, and it was confirmed that all cells could be cultured more than the initial inoculation amount and the culture reproducibility. In addition, it was confirmed that the differentiated cells of each stage of the present invention can function as sufficient infection hosts, as the number of HAV gene copies increased after 3 days after inoculation. And thus, by inoculating HAV at each stage of differentiation and selecting the optimal time point of culture, it is possible to shorten the culture period and establish an efficient culture system with optimal effect.

The invention relates to the field of biomedical engineering and disease model construction, in particular to a construction method and application of a full-spectrum liver organoid. The construction method comprises the following steps: respectively differentiating induced pluripotent stem cells from human umbilical cord to obtain transitional hepatic endoderm cells, endothelial progenitor cells, red myeloid progenitor cells, hepatic stellate cells and diaphragm mesenchymal stem cells; transition hepatic endoderm cells, endothelial progenitor cells, red myeloid progenitor cells, hepatic stellate cells and diaphragm mesenchymal stem cells are inoculated into a 3D culture system according to the cell number ratio of (9-11): (6-8): (1-2): (1-2): (1-2); and performing self-assembly co-culture on the seed cells in a 3D culture system until the liver function is mature, thereby obtaining the full-spectrum liver organoid. The technical problem that physiological authenticity, functional integrity and application universality of an existing liver model need to be improved is solved.

Provided herein are 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. 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.

The invention relates to the technical field of biology, and particularly discloses a method for inducing front anterior intestine endoderm cells to differentiate to form lung progenitor cells and application of the method. The method for inducing anterior front intestine endoderm cells to differentiate to form the lung progenitor cells comprises the step of inducing anterior front intestine endoderm single cells into the lung progenitor cells by using a lung progenitor cell culture medium in a solidified 3D culture composite adhesive, the 3D culture composite adhesive is prepared from 3D culture hydrogel and polydopamine nanoparticles modified by polyethylene glycol. The cell three-dimensional culture system constructed by mixing the PEG-PDA NPs and the 3D culture hydrogel can promote differentiation of the human pluripotent stem cells to the lung progenitor cells, the differentiation efficiency and quality are improved, and a good foundation is laid for subsequent efficient formation of mature lung organs with trachea and alveolar characteristics.

This disclosure provides cell-based compositions for the treatment of diabetes, methods for identifying cells that preferentially differentiate into endoderm cells, and methods for preparing insulin-producing pancreatic cells, as well as related methods of use for the treatment of insulin deficiency-related diseases.

The invention discloses a universal type definitive endoderm culture medium for inducing pluripotent stem cells to differentiate into definitive endoderm cells and a culture method, and relates to the technical field of stem cell biology and regenerative medicine. The universal typing endoderm culture medium comprises a basic culture medium, polyvinyl alcohol, an animal-origin-free B-27 supplement, a TGF-beta activator, a Wnt signaling pathway activator, a JNK-JUN inhibitor, a PI3K / mTOR inhibitor, L-ascorbic acid and a ROCK inhibitor, wherein the basic culture medium is used for providing nutritional ingredients required by cell growth; the universal typified endoderm culture medium comprises a typified endoderm culture medium 1 and a typified endoderm culture medium 2, according to the culture method, the general type typing endoderm culture medium is constructed to obtain the typing endoderm cell balls, and the general type typing endoderm culture medium is definite in component and does not contain animal-derived components, so that the general type typing endoderm culture medium meets the requirement of a drug supervision institution on chemical component limitation of cell treatment products; high-density culture is carried out on the typified endoderm cells through suspension culture, so that the method is more suitable for production requirements.

To provide a method for efficiently inducing differentiation in vitro of spherical human cell aggregates containing axial mesodermal cells from human pluripotent stem cells, in order to realize the transplant application to patients with degenerative disc disease; and to provide the spherical human cell aggregates containing axial mesodermal cells induced to differentiate in vitro.SOLUTION: A spherical human cell aggregate has the following characteristics: (1) it has two distinct regions, a primitive node area and a non-node area; (2) the primitive node area and the non-node area have a structure consisting of at least two layers; (3) axial mesoderm cells are present in the first layer on the surface of the primitive node area, and endoderm cells are substantially absent in the primitive node area; and (4) endoderm cells are present in the first layer on the surface of the non-node area.SELECTED DRAWING: None

The invention relates to a cellulose derivative-based hydrogel, the matrix of which comprises a cellulose derivative, the matrix comprises a net structure composed of ultramicro fibers and has a highly-ordered three-dimensional micro-nano structure, and the net structure composed of ultramicro fibers has a highly-ordered three-dimensional micro-nano structure. The cellulose derivative-based hydrogel is a ready-to-use hydrogel which does not contain animal-derived components and has no toxicity to cells, and is suitable for 3D cell culture, tissue and organoid culture, stem cell culture and organoid induced differentiation. The method has the advantages that dense fused monolayer endoderm cells can be formed, non-directional differentiation of stem cells can be inhibited, and formed organoid vesicles have thick and solid vesicles walls, are mature organoid differentiation and can be used for high-throughput drug screening, drug toxicity testing and regenerative medicine.