15 results about "Definitive endoderm" patented technology

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.

A method and system are disclosed for differentiating non-human primate induced pluripotent stem cells (NHP iPSCs) into hepatocyte-like cells through a defined, multi-stage protocol. The method includes sequentially exposing NHP iPSCs to six distinct cell differentiation media over an approximate eighteen-day period. Each medium comprises defined combinations of growth factors, small molecules, and inhibitors that direct progressive lineage specification through anterior primitive streak, definitive endoderm, posterior foregut, liver bud progenitors, hepatoblasts, and hepatocyte-like cells. Concentration ranges and exposure durations are provided for each stage, ensuring reproducibility and reducing heterogeneity across cell populations. In some embodiments, treatment with a transforming growth factor beta (TGF-β ) receptor I inhibitor during early hepatoblast specification enhances lineage fidelity. The resulting hepatocyte-like cells are validated by immunostaining and western blotting for hepatic markers. Applications include preclinical drug metabolism, hepatotoxicity assessment, and in vitro disease modeling using non-human primate–derived systems.

The present disclosure provides thymic cells, thymic organoids, and animal models comprising such thymic cells and thymic organoids. The present disclosure also provides methods for inducing, in vitro and in vivo, differentiation of pluripotent stem cells into definitive endoderm (DE), anterior and anterior intestinal endoderm (AFE), ventral pharyngeal endoderm (VPE), thymic epithelial progenitor (TEP) cells, thymic epithelial cells (TEC), and thymic organoid.

The invention relates to methods of pluripotent stem cell (PSC) culture, PSC cultured in 3D suspension culture bioreactor systems, expansion of PSC in 3D suspension, and differentiation of 3D expanded PSC clusters into Definitive Endoderm (DE).

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.

The invention discloses a method for culturing vascularization of interstitial-intestinal organs, and belongs to the technical field of biology. The method comprises the following steps: inducing deterministic endoderm; inducing a posterior intestinal endoderm: digesting into single cells and transferring the single cells into an agarose microarray to polymerize into balls; and inducing vascularization of intestinal organs. According to the method for realizing co-differentiation after aggregation of the agarose microarray, the size of the initial intestinal spheroid can be regulated and controlled by adjusting the size of the microarray, and the size of the initial intestinal spheroid is ensured to be consistent; by adjusting the initial cell state, the rapid growth of intestinal organs can be realized, and the growth rate of the polymerized balls is far greater than that of single cells; based on the microarray technology, the problem that the regeneration efficiency is low due to the fact that the spontaneous balling rate of a traditional method is low is solved, the extremely high effective balling rate is achieved, and the intestinal organs with more developed interstitial tissues and the higher vascularization degree are cultivated.

Disclosed is an in-vitro protocol for differentiating human induced pluripotent stem cells (hiPSCs) or human embryonic stem cells (hESC) to give rise to a definitive endoderm, followed by progression into anteriorized foregut endoderm that has the ability to give rise to both proximal and distal lung epithelial cells. The protocol not only offers great opportunities for the study of human development but also have tremendous potential for future clinical cell-based therapies. The protocol outlined here is used to differentiate hiPSCs into lung epithelial cell types through a process that faithfully recapitulates the stepwise events observed in-vivo. The was followed with the working cell bank of an hiPSC line made under current Good Manufacturing Practice (cGMP) conditions, a necessary step for the future clinical application of these cells.

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

Disclosed is an in-vitro protocol for differentiating human induced pluripotent stem cells (hiPSCs) or human embryonic stem cells (hESC) to give rise to a definitive endoderm, followed by progression into anteriorized foregut endoderm that has the ability to give rise to both proximal and distal lung epithelial cells. The protocol not only offers great opportunities for the study of human development but also have tremendous potential for future clinical cell-based therapies. The protocol outlined here is used to differentiate hiPSCs into lung epithelial cell types through a process that faithfully recapitulates the stepwise events observed in-vivo. The was followed with the working cell bank of an hiPSC line made under current Good Manufacturing Practice (cGMP) conditions, a necessary step for the future clinical application of these 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 preparation method and application of prostate lineage progenitor cells obtained by multi-stage induction of human induced pluripotent stem cells. The method comprises the following steps: sequentially inducing to form a definitive endoderm, a posterior intestinal urogenital sinus progenitor cell and a prostate progenitor cell, and at the third stage, adopting a collaborative regulation system in which LDN193189 and CHIR99021 are simultaneously added, so that the lineage specialization efficiency and the cell amplification capacity are remarkably improved. And further sorting through a CD49f high-expression membrane marker combination, a Trop2 positive membrane marker combination, a CD26 low-expression membrane marker combination and a CD117 negative membrane marker combination, so that a cell population with high purity, low heterogeneity and immunoregulation potential can be obtained. The cell can reconstruct a gland structure and secrete prostate specific protein in an animal model, and meanwhile, the inflammatory microenvironment is improved through immune tolerance related pathways. The process can realize batch production and maintain product activity and stability, and is suitable for treating chronic prostatitis, postoperative tissue defect, dysfunction and other 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.

Among the various aspects of the present disclosure is the provision of methods for the generation of stem cell-derived beta cells (SC-β), stem cell-derived islets (SC-islets), definitive endoderm cell, pancreatic progenitor cells, and uses thereof. The various cells are generated by contacting human pluripotent stem cells (hPSCs) with an amount of a cytoskeleton depolymerizing agent during initial stem cell differentiation into definitive endoderm cells. A method of treatment of a subject diagnosed with a diabetes condition by transplanting administering a therapeutically effective amount of the stem cell-derived islets (SC-islets) into the subject is also disclosed.